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PRODUCT SPECIFICATIONS:
Tissue: Positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain: Fontana Masson quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

CONTROL SLIDE VALIDATION:
With Fontana Masson Stain Kit: Part 9105A Individual Stain Solution
Solution A: Silver Nitrate 10%, Aqueous 250 ml Part 13806
Solution B: Ammonium Hydroxide 28-30%, ACS 250 ml Part 1006
Solution C: Gold Chloride 0.2%, Aqueous 250 ml Part 11286
Solution D: Sodium Thiosulfate 5%, Aqueous 250 ml Part 1389
Solution E: Nuclear Fast Red Stain, Kernechtrot 250 ml Part 1255

APPLICATION:

Newcomer Supply Argentaffin Control Slides are for the positive histochemical staining of argentaffin substances in tissue sections.

PRESTAINING PREPARATION:

1. Heat dry sections in oven according to your laboratory protocol.
2. All glassware/plasticware must be acid cleaned prior to use.
a. See Procedure Notes #1 and #2.
3. Prepare Fontana Silver Working Solution (diamine silver) in an acid cleaned Erlenmeyer flask:
a. Solution A: Silver Nitrate 10%, Aqueous; 25 ml
b. Add Solution B: Ammonium Hydroxide 28-30%, ACS drop by drop, mix with swirling motion until solution clouds, then clears. Use caution to not add too much Solution B: Ammonium Hydroxide 28-30%, ACS.
c. Add more Solution A: Silver Nitrate 10%, Aqueous drop by drop until clear solution becomes slightly cloudy.
d. Let solution stand for 2-4 hours before use.
e. For use; after standing, filter the solution. Combine 20 ml of this filtered diamine silver solution with 40 ml of distilled water; 60 ml total.

NEWCOMER SUPPLY VALIDATION PROCEDURE:

4. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each. Wash well with distilled water.
a. See Procedure Notes #3 and #4..
5. Immerse slides in the Fontana Silver Working Solution (Step #3) in a 45°C to 60°C water bath for 1 hour.
6. Check slides microscopically; remove control, rinse in warm distilled water. Confirm that reaction is complete when granules are dark brown and background is colorless.
a. Return to heated Fontana Silver Working Solution for longer incubation if indicated.
7. Rinse well in three changes of distilled water.
8. Immerse in Solution C: Gold Chloride 0.2%, Aqueous; 10 minutes.
9. Rinse well in distilled water.
10. Place in Solution D: Sodium Thiosulfate 5%, Aqueous; 5 minutes.
11. Rinse well in distilled water.
12. Counterstain in Solution E: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
a. Shake solution well before use; do not filter.
13. Rinse well in distilled water.
a. See Procedure Note #5.
14. Dehydrate quickly in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

CaviWipes™ Bleach disinfecting towelettes are non-woven disposable towelettes  pre-saturated with CaviCide™ Bleach.

With patent pending Enhanced Chlorine Technology™ (ECT™) they have demonstrated effectiveness against 41 relevant microorganisms – including C. diff spores in 3 minutes when used as directed on hard, non-porous surfaces.

They kill C. diff spores in 3 minutes—and do not require PPE*. They also meet the CDC recommended 1:10 bleach dilution and are compatible with most medical device equipment.  So when it comes to disinfecting hard non-porous surfaces, leave the gloves behind and reach for CaviWipes Bleach.

    • Kill all product labeled organisms in 3 minutes including C. diff, TB, drug resistant bacteria, viruses and fungi
    • No PPE required*
    • 1:10 Bleach dilution – Compatible with most medical device equipment**
    • One-step cleaner and disinfectant
    • Low Odor

 

*Special instructions for Cleaning Prior to Disinfection against Clostridium difficile spores, HIV-1, HBV, HCV.

CaviWipes Bleach Directions
CaviWipes Bleach Technical Bulletin

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal spleen and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  AFB, Fite quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

CONTROL SLIDE VALIDATION:

With AFB, Fite Stain Kit: Part 91013A Individual Stain Solution
Solution A:  Xylene/Peanut Oil, 2:1 500ml Part 1449
Solution B:  Carbol Fuchsin Stain, Ziehl-Neelsen 250ml Part 1030
Solution C:  Acid Alcohol 1% 250ml Part 10011
Solution D:  Light Green SF Yellowish 0.1%, Aqueous 250ml Part 12203

 

APPLICATION:

Newcomer Supply Fite, Leprosy, Animal Control Slides are for the positive histochemical staining of Mycobacterium leprae sp., the causative agent of leprosy.

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Filter Solution B: Carbol Fuchsin Stain, Ziehl-Neelsen with high quality filter paper.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

      1. Deparaffinize slides in Solution A: Xylene/Peanut Oil, 2:1, two changes for 10 minutes each.
        1. See Procedure Note #1.
      2. Drain slides, wipe off excess oil, and blot to opacity taking care to remove residual oil.
        1. See Procedure Note #2.
      3. Stain in freshly filtered Solution B: Carbol Fuchsin Stain, Ziehl-Neelsen for 15 minutes at room temperature.
      4. Rinse well in distilled water.
      5. Differentiate slides individually in Solution C: Acid Alcohol 1% until sections are light pink; 5-10 dips.
      6. Rinse well in distilled water.
      7. Counterstain in Solution D: Light Green SF Yellowish 0.1%, Aqueous; 5-10 dips.
      8. Rinse in distilled water.
      9. Blot excess water from slide and air-dry or oven-dry completely.
      10. Dip dried slides in xylene and coverslip with a compatible mounting medium.

 

RESULTS:

Mycobacterium leprae Red
Other tissue elements Green
Negative lung Negative for Mycobacterium leprae

 

PROCEDURE NOTES:

    1. Acid-fastness of leprosy organisms is enhanced when the waxy capsule is protected by the mixture of xylene/peanut oil and avoidance of dehydrating solutions.
    2. It is important to blot well, residual oil may produce staining artifact.
    3. If using a xylene substitute, closely follow the manufacturer’s recommendations for coverslipping step.

 

REFERENCES:

    1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 220-221.
    2. Fite, George, P.J. Cambre and M.H. Turner. “Procedure for Demonstrating Lepra Bacilli in Paraffin Sections”. Archives of Pathology 43 (1947). 624-625.
    3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
    4. Modifications developed by Newcomer Supply Laboratory.

Actin, Muscle Specific, Control Slides contain a section of positive staining uterus.

 

PRODUCT DESCRIPTION:

The enclosed positive control slides are intended to be used to verify histological techniques and reagent reactivity.  These slides are to be used for the qualitative purpose of determining positive or negative results, and are not intended to be used for any quantitative purpose.  The first serial section within the control box is stained and provided for your reference.  Before using the unstained slides, review the enclosed stained slide with your pathologist to ensure that this tissue source is acceptable.  Newcomer Supply will not accept a return with missing slides in the series. Newcomer Supply guarantees reactivity of these control slides for one year from the date of receipt.  Revalidate after one year to verify continued reactivity.  Store at 15-30°C in a light deprived and humidity controlled environment.

These positive control slides were produced from human surgical or autopsy tissues under carefully controlled conditions.  Reasonable measures are used to deliver quality control slides that are as consistent as possible.  However, characteristics of quality control slides may be dissimilar due to variations in the reagents, stains, techniques, laboratory conditions, and tissue sources used. Newcomer Supply Laboratory uses a manual method of performing quality control procedures, specifically avoiding automation, in order to provide reactive control slides for even less aggressive methods of staining that our customers may be using.

 

APPLICATION:

Newcomer Supply Actin, Muscle Specific, Control Slides are for the positive immunohistochemical staining of actin, a major component of the cytoskeleton, present in skeletal, cardiac and smooth muscle cells.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique: Paraffin sections cut at 4 microns on Superfrost® Plus

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  2. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  3. Rinse in distilled water; tap off excess water.
  4. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  5. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  6. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  7. Tap off excess buffer; apply Actin, Muscle Specific primary antibody.  Incubate at room temperature for 30 minutes.
  8. Rinse slides in two changes of buffer.
  9. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  10. Rinse slides in two changes of buffer.
  11. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  12. Rinse slides in two changes of buffer.
  13. Prepare required quantity of DAB substrate/chromogen.
  14. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  15. Rinse slides in four changes of distilled water.
  16. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  17. Rinse slides in warm tap water to blue sections.
  18. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Actin, Muscle Specific positive expression Brown cytoplasmic staining
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during staining procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Actin, Muscle Specific (HHF35) is the pre-diluted primary antibody used.
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Actin, Muscle Specific Antibody datasheet.
  2. Cell Marque HiDef Detection™ Polymer System datasheet.
  3. Cell Marque DAB Substrate Kit datasheet.
  4. Modifications developed by Newcomer Supply Lab

Solution A: Conditioning Solution A

Solution B: Conditioning Solution B

Solution C: FrozFix® Fixative
Solution D: Tris Buffered Saline + Tween, Mod., pH7.6, 10X

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal tissue and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Brown-Hopps quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Gram, Brown-Hopps Stain Kit: Part 9124A Individual Stain Solution
Solution A:  Crystal Violet Stain 1%, Aqueous, Brown-Hopps 250 ml Part 1041
Solution B:  Iodine, Gram, Aqueous 250 ml Part 1140
Solution C:  Basic Fuchsin Stain 0.25%, Aqueous 250 ml Part 1011
Solution D:  Gallego Solution 250 ml Part 1098
Solution E:  Picric Acid-Acetone 0.05% 250 ml Part 13351
Solution F:  Acetone-Xylene 1:1 250 ml Part 10015
Acetone, ACS Part 10014

 

APPLICATION:

Newcomer Supply Gram Positive & Gram Negative Bacteria, Animal Control Slides are for the positive histochemical staining of gram positive and gram negative bacteria in a naturally occurring infection.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Stain in Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps for 2 minutes.
  4. Rinse well in distilled water, ensuring excess stain is removed.
  5. Mordant in Solution B: Iodine, Gram, Aqueous for 2 minutes.
    1. Sections should turn black.
  6. Rinse well in running tap water, ensuring excess iodine is removed.
  7. Blot excess water from slide; decolorize one slide at a time in Acetone, ACS (Part 10014) until blue color stops running; 1-2 dips.
    1. Sections should be very light gray in color.
  8. Quickly rinse in running tap water to remove excess Acetone.
  9. Stain in Solution C: Basic Fuchsin Stain 0.25%, Aqueous; 5 minutes.
  10. Rinse well in running tap water.
  11. Differentiate in Solution D: Gallego Solution for 5 minutes.
  12. Rinse in running tap water.  Blot water off slide(s), but not to dryness.
    1. Proceed with Steps #13 to #16 one slide at a time.
  13. Dip quickly in Acetone, ACS (Part 10014); 1-2 dips.
  14. Dip directly in Solution E: Picric Acid-Acetone 0.05%; 3-10 dips.
  15. Dip quickly in Solution F: Acetone-Xylene 1:1; 5 dips.
  16. Clear in three changes of xylene, 10 dips each; coverslip with  compatible mounting medium.

 

RESULTS:

Gram positive bacteria Blue/violet
Gram negative bacteria Red
Nonreactive lung Negative for gram positive/negative bacteria

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Brown, Robert C., and Howard C. Hopps. “Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.” American Journal of Clinical Pathology 60.2 (1973): 234-240.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 222-224.
  3. Modifications developed by Newcomer Supply Laboratory.

 

 

 

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining submandibular gland and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  GCDFP-15 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply GCDFP-15 (gross cystic disease fluid protein 15) Control Slides are for the positive immunohistochemical staining of GCDFP-15, found in cyst fluid of cystic breast disease and a marker for primary and metastatic breast carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply GCDFP-15 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

GCDFP-15 positive expression Brown cytoplasmic staining
Myometrium Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque GCDFP-15 (23A3) is the concentrated primary antibody used.  Dilute primary antibody to 1/150 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque GCDFP-15 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

   

PERIODIC ACID SCHIFF (PAS) STAIN KIT INCLUDES: 

Part 9162A Part 9162B
Solution A: Periodic Acid 0.5%, Aqueous 250 ml 500 ml
Solution B: Schiff Reagent, McManus 250 ml 500 ml
Solution C: Hematoxylin Stain, Harris 250 ml 500 ml
Solution D: Acid Alcohol 1% 250 ml 500 ml
Solution E: Lithium Carbonate, Saturated Aqueous 250 ml 500 ml

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed are two complimentary unstained positive control slides for initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers at www.newcomersupply.com.

Additionally Needed:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Alpha Amylase 1%, Aqueous Part 1905  (for glycogen digestion)
Coplin Jar, Plastic Part 5184  (for glycogen digestion microwave modification)

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Periodic Acid Schiff (PAS) Stain Kit procedure, with methods for glycogen digestion, is used for staining glycoproteins and may aid in the differential diagnosis of tumors through detection of acid/neutral epithelial mucins and/or glycogen.   Digestion steps can be employed for further identification of mucosubstances.  PAS can also be used for staining basement membranes and fungal cell walls.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
    2. Proceed to Step #5 if not running Digestion.
  3. Digestion Step:  See Procedure Note #3.
    1. Two control slides and two patient slides are needed.
    2. Label one control slide and one patient slide “with”.
    3. Label the other control slide and patient slide “without”.
    4. Place slides labeled “without” in separate Coplin jar of distilled water; hold for Step #5.
    5. Apply Alpha Amylase 1%, Aqueous (Part 1905) to slides labeled “with” for 30 minutes at room temperature.        
    6. Proceed to Step #5.
  4. Digestion Microwave Modification:  See Procedure Note #4.
    1. Follow Steps #3a through #3d.
    2. Place slides labeled “with” in a plastic Coplin jar containing Alpha Amylase 1%, Aqueous (1905) and microwave  for 1 minute at 37°C. Let sit in warm solution for an additional minute.
  5. Combine all slides for remaining steps;  wash in running tap water for 1 minute, rinse in distilled water.
  6. Place in Solution A: Periodic Acid 0.5%, Aqueous for 10 minutes.
  7. Wash in three changes of tap water; rinse in distilled water.
  8. Place in Solution B: Schiff Reagent, McManus for 20 minutes.
  1. Wash in lukewarm tap water for 5 minutes.
  2. Stain with Solution C: Hematoxylin Stain, Harris 1-5 minutes, depending on preference of nuclear stain intensity.
  3. Wash in tap water for 2-3 minutes.
  4. Differentiate in Solution D: Acid Alcohol 1%; 1-2 quick dips.
  5. Wash in tap water for 1 minute.
  6. Blue in Solution E: Lithium Carbonate, Saturated Aqueous; 3-4 dips.
  7. Wash in several changes of tap water; rinse in distilled water.
  8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Glycogen  Magenta
Glycogen digestion Absence of magenta
Acid & neutral epithelial mucin Magenta
Fungal cell walls Red to purple
Basement membranes Red to purple
Nuclei Blue

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Slides labeled “with” will be treated with amylase digestion, slides labeled “without” will not be treated for digestion.
  4. The suggested microwave procedure has been tested at Newcomer Supply.  This procedure is a guideline and techniques should be developed for use in your laboratory.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-171, 180.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168, 245.
  4. Modifications developed by Newcomer Supply Laboratory.

 

  

Newcomer Supply PAP GnomePen Liquid Blocker creates a thin visible hydrophobic film around tissue sections to reduce the amount of reagent needed for tissue coverage and stain reaction. GnomePen has one of the thinnest PAP pen tips available, allowing for adjacent circles, lines or a drawn pattern to be applied on a single slide.

 

GnomePen contains a unique formulation that is water repellent, insoluble in alcohol and acetone, soluble in xylene and is insensitive to detergents (Triton X-100, Tween 20), varying pH and temperature. Two barrier colors are available for color coding and specimen differentiation on the same slide. Each GnomePen contains approximately 350 applications in a transparent container that provides an accurate visual estimate of remaining solution volume.

 

METHOD:

Technique:  Paraffin, frozen sections and tissue culture cells

      • Manual staining for:
        • Immunohistochemistry (IHC) procedures
        • Immunofluorescence Assay (IFA) procedures

 

PROCEDURE:

 

      1. Shake pen thoroughly before use.
      2. Practice applying GnomePen liquid barrier on a test slide; push pen tip against the glass and apply a thin liquid barrier that dries to a film. If flow does not instantly begin, gently squeeze the pen body.
      3. Store GnomePen tightly capped; vertically with cap end up.
      4. Paraffin Section Method:
        1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        2. Remove slide from water or buffer; blot excess solution from slide and tissue section. Or place long edges of slide on absorbent material to remove excess moisture.
        3. Encircle tissue section(s) on slide surface with GnomePenas illustrated. Do not touch the pen to any edge of the tissue.
        4. The liquid barrier can be drawn on a slightly damp slide.
        5. See Procedure Notes #1 and #2.
      5. Frozen Section and Tissue Culture Cells:
        1. Encircle frozen tissue section(s) or tissue culture cells on slide surface at room temperature with the GnomePen as illustrated. Do not touch the pen to any edges of the tissue or culture cells.
        2. See Procedure Notes #1 and #2.
        3. The liquid barrier should be applied before fixation or prior to the immersion of slide into water or buffer.
      6. After liquid barrier application, allow slide to dry in a flat position for 30-60 seconds at room temperature. Proceed with procedure when the drawn barrier has completely dried.
        1. See Procedure Note #3.
      7. Drain/rinse reagents off between staining steps; blotting slide and tissue as needed to remove excess solution.
        1. See Procedure Note #4.
      8. Complete staining and coverslip with a compatible mounting medium.

 

PROCEDURE NOTES:

 

      1. Once a tissue section is touched with GnomePen liquid barrier it cannot be removed. The section remains useable but slight colorization will result on the touched tissue.
      2. If the tissue section is not completely encircled or segregated by the GnomePen barrier film, reagents will not be fully retained on the tissue section and flood out onto the slide. This may compromise complete and adequate tissue coverage by reagents.
      3. If GnomePen barrier lines are not completely dry prior to staining, a precipitate from reaction with detection reagents may occur.
      4. The use of a Slide Moisture Chamber or StainTray™ (Part 68431, 6848 or 6847) is recommended for manual staining to maintain slide organization and a moist environment during the procedure.

 

REFERENCES:

 

      1. Grizzle, William, Cecil Stockard, and Paul Billings. “The Effects of Tissue Processing Variables Other Than Fixation on Histochemical Staining and Immunohistochemical Detection of Antigens.” The Journal of Histotechnology3 (2001): 213-219.
      2. Modifications developed by Newcomer Supply Laboratory.

 

 

Lymph node reactive for Anaplastic Lymphoma Kinase (ALK, CD246).

A percentage of patients with Anaplastic Large Cell Lymphoma (ALCL) or Non Small Cell Lung Cancer (NSCLC) will express ALK.

Retrieve easily and gently at 60°C for 30 minutes or 75°C for 15 minutes. Low temperature retrieval solution universally applicable to all antibodies and tissues. Also for IHC, Immunofluorescence, DNA & RNA probes. No Boiling. No High pH Buffer. No Citrate. Contains No Enzymes.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  PAS quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Periodic Acid Schiff (PAS) Stain Kit: Part 9162A/B Individual Stain Solution
Solution A: Periodic Acid 0.5%, Aqueous 250/500 ml Part 13308
Solution B:  Schiff Reagent, McManus 250/500 ml Part 1371
Solution C:  Hematoxylin Stain, Harris 250/500 ml Part 12013
Solution D:  Acid Alcohol 1% 250/500 ml Part 10011
Solution E:  Lithium Carbonate, Saturated Aqueous 250/500 ml Part 12215
Alpha Amylase 1%, Aqueous (for glycogen digestion)   Part 1905
Coplin Jar, Plastic (for glycogen digestion microwave modification) Part 5184

 

APPLICATION:

Newcomer Supply Periodic Acid Schiff (PAS) Glycogen Control Slides are for the positive histochemical staining of glycogen in tissue sections and can also be utilized as control slides for glycogen digestion steps.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
    2. Proceed to Step #5 if not running Digestion
  3. Digestion Step:  See Procedure Note #3.
    1. Two control slides and two patient slides are needed. 
    2. Label one control slide and one patient slide “with”.
    3. Label the other control slide and patient slide “without”.
    4. Place slides labeled “without” in separate Coplin jar of distilled water; hold for Step #5.
    5. Apply Alpha Amylase 1%, Aqueous (Part 1905) to slides labeled “with” for 30 minutes at room temperature.    
    6. Proceed to Step #5.
  4. Microwave Modification:  See Procedure Note #4.
  1. Follow Steps #3a through #3d.
  2. Place slides labeled “with” in a plastic Coplin jar containing Alpha Amylase 1%, Aqueous (Part 1905) and microwave  for 1 minute at 37°C. Let sit in warm solution for an additional minute.
  1. Combine all slides for remaining steps; wash in running tap water for 1 minute; rinse in distilled water.
  2. Place in Solution A: Periodic Acid 0.5%, Aqueous for 10 minutes.
  3. Wash in three changes of tap water; rinse in distilled water.
  4. Place in Solution B: Schiff Reagent, McManus for 20 minutes.
  5. Wash in lukewarm tap water for 5 minutes.
  6. Stain with Solution C: Hematoxylin Stain, Harris, 1 to 5 minutes, depending on preference of nuclear stain intensity.
  7. Wash in tap water for 2-3 minutes.
  8. Differentiate in Solution D: Acid Alcohol 1%; 1-2 quick dips.
  9. Wash in tap water for 1 minute.
  10. Blue in Solution E: Lithium Carbonate, Saturated Aqueous; 3-4 dips.
  11. Wash in several changes of tap water; rinse in distilled water.
  12. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Glycogen digestion Absence of magenta
Glycogen Magenta
Acid & neutral epithelial mucin Magenta
Nuclei Blue

 

  PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Slides labeled “with” will be treated with amylase digestion, slides labeled “without” will not be treated for digestion.
  4. The suggested microwave procedure has been tested at Newcomer Supply.  This procedure is a guideline and techniques should be developed for use in your laboratory.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-171, 180.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
  3. Sheehan, Dezna C., and Barbara B.Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168.
  4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Gordon & Sweets Reticulum quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Reticulum, Gordon & Sweets Stain Kit: Part 9168A Individual Stain Solution
Solution A: Potassium Permanganate 1%, Aqueous 250 ml Part 13393
Solution B: Oxalic Acid 1%, Aqueous 250 ml
Solution C: Ferric Ammonium Sulfate 2.5%, Aqueous 250 ml
Solution D: Silver Nitrate 10%, Aqueous 50 ml Part 13806
Solution E: Ammonium Hydroxide 28-30%, ACS 50 ml Part 1006
Solution F: Sodium Hydroxide 3%, Aqueous 50 ml
Solution G: Formalin 10%, Aqueous 250 ml
Solution H: Gold Chloride 0.2%, Aqueous 250 ml Part 11286
Solution I: Sodium Thiosulfate 5%, Aqueous 250 ml Part 1389
Solution J: Nuclear Fast Red Stain, Kernechtrot 250 ml Part 1255

 

APPLICATION:

Newcomer Supply Reticulum Control Slides are for the positive histochemical staining of reticulum fibers; regarded as specialized connective tissue fibers.

 

PRESTAINING PREPARATION:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. All glassware/plasticware must be acid cleaned prior to use.
    1.  See Procedure Notes #1 and #2.
  3. Prepare Ammoniacal Silver Working Solution.  Save for Step #11.
    1. Place 5 ml of Solution D: Silver Nitrate 10%, Aqueous in a flask.
    2. Add Solution E: Ammonium Hydroxide 28-30%, ACS drop by drop, continuously swirling until formed precipitate completely dissolves.  Do not add any excess Ammonium Hydroxide. 
    3. Add 5 ml of Solution F: Sodium Hydroxide 3%, Aqueous.
    4. Re-dissolve formed precipitate with Solution E: Ammonium Hydroxide 28-30%, ACS until a faint cloudiness remains.
    5. If proceeded too far and no cloudiness remains, add Solution D: Silver Nitrate 10%, Aqueous drop by drop, until one drop causes solution to become permanently cloudy. Faint cloudiness is the optimum.
    6. Bring solution volume to 50 ml with distilled water; filter.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #3 and #4.
  2. Oxidize in Solution A: Potassium Permanganate 1%, Aqueous for 3 minutes.
  1. Wash in running tap water for 1 minute; rinse in distilled water.
  2. Bleach in Solution B: Oxalic Acid 1%, Aqueous for 2 minutes or until sections are colorless.
  3. Wash in running tap water for 1 minute; rinse in distilled water.
  4. Sensitize in Solution C: Ferric Ammonium Sulfate 2.5%, Aqueous; 10 to 15 minutes.
  5. Rinse in several changes of distilled water.
  6. Impregnate sections in filtered Ammoniacal Silver Working Solution (Step #3) for 2 minutes.
  7. Rinse well in running distilled water for 1 minute.
    1. See Procedure Note #5.
  8. Reduce in Solution G: Formalin 10%, Aqueous for 1 minute.
  9. Rinse in running tap water for 1 minute.
  10. Check control slide microscopically for sufficient black reticular fiber development.
    1. See Procedure Note #6.
  11. Tone in Solution H: Gold Chloride 0.2%, Aqueous for 1-2 minutes.
  12. Rinse well in distilled water.
  13. Place in Solution I: Sodium Thiosulfate 5%, Aqueous for 1 minute.
  14. Wash well in tap water for 1 minute; rinse in distilled water.
  15. Counterstain with Solution J: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
    1. Shake solution well before use; do not filter.
  16. Rinse well in distilled water.
    1. See Procedure Note #7.
  17. Quickly dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Reticular fibers Black
Background Red

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
  2. Plastic (Part 5500), plastic-tipped or paraffin coated metal forceps must be used with silver solutions to prevent precipitation of silver salts.  No metals of any kind should come in contact with silver solutions.
  3. Drain slides after each step to prevent solution carry over.
  4. Do not allow sections to dry out at any point during procedure.
  5. This rinse step is critical for good reticulum demonstration.  If rinsing is insufficient, excessive background staining may occur.
  6. If black reticular fibers are not evident or are lightly/poorly stained, return all slides to Ammoniacal Silver Working Solution (Step #11) and repeat Steps 11-14 with the same timings.
  7. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
  8. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 177-179.
  2. Gordon, Harold, and Henry Sweets. “A Simple Method for the Silver Impregnation of Reticulum.” American Journal of Pathology 12.4 (1936): 545-552.
  3. Modifications developed by Newcomer Supply Laboratory.

  

Safety Overview – Click Here

FUNGUS STAIN KIT INCLUDES:  

Part 9121A Part 9121B
Solution A: Chromic Acid 5%, Aqueous 250 ml 500 ml
Solution B: Sodium Bisulfite 1%, Aqueous 250 ml 500 ml
Solution C: Silver Nitrate 125 ml 250 ml
Solution D: Methenamine Borate 125 ml 250 ml
Solution E: Gold Chloride 0.1%, Aqueous 250 ml 500 ml
Solution F: Sodium Thiosulfate 2%, Aqueous 250 ml 500 ml
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250 ml 500 ml

  

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed are two complimentary unstained positive control slides for the initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers.

 

Additionally Needed:

Hydrochloric Acid 5%, Aqueous Part 12086 (for acid cleaning glassware)
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Coplin Jar, Plastic Part 5184 (for microwave modification)

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Fungus, Grocott Methenamine Silver (GMS) Stain Kit procedure, with included microwave modification, is one of the best staining methods to demonstrate a variety of fungal organisms including: Pneumocystis, Aspergillus, Blastomyces, Candida and Histoplasma.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns.

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. All glassware/plasticware must be acid cleaned prior to use.
    1. See Procedure Notes #1 and #2.
  3. Prepare Silver-Methenamine Working Solution and mix well.
    1. Solution C: Silver Nitrate                      20 ml
    2. Solution D: Methenamine Borate 20 ml
  4. Preheat Silver-Methenamine Working Solution to 45°C-60°C in a water bath approximately 20 to 30 minutes before use.
    1. See Procedure Note #3.
    2. Do not preheat if using Microwave Modification; Step 11.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #4 and #5.
  2. Oxidize in Solution A: Chromic Acid 5%, Aqueous for 1 hour.

        Microwave Modification: See Procedure Note #6

    1. Oxidize slides in a plastic Coplin jar containing Solution A:

       Chromic Acid 5%, Aqueous and microwave for 1 minute and 20 seconds at 60°C.

  1. Wash well in running tap water; rinse in distilled water.
  2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
  3. Wash for 5 minutes in running tap water; rinse well in distilled water.
  4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°C-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
    1. Periodically remove control, rinse in warm distilled water, check microscopically for adequate silver impregnation.  Fungi should be dark brown. 
    2. If organisms are not sufficiently dark, return slides to warm silver solution.  Recheck at 2-3 minute intervals until desired intensity is achieved.
    3. Pneumocystis may take longer to stain than other fungus.
    4. Staining at room temperature will require longer incubation.
  5. Microwave Modification:
    1. Incubate slides in a plastic Coplin jar containing Silver-Methenamine Working Solution and microwave for 1 minute at 70°C.
    2. Check microscopically for adequate development.
    3. If additional incubation is required, return slides to warm silver solution.  Recheck at 2-3 minute intervals.
  6. Rinse in three to four changes of distilled water.
    1. Never use tap water at this step.
  7. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 20 seconds to 1 minute.
  8. Rinse well in distilled water.
  9. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
  10. Wash in running tap water for 5 minutes; rinse in distilled water.
  11. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
  12. Dehydrate quickly in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Fungi Crisp black cell walls with visible internal structures
Background Green
Mucin Taupe to dark gray

 

PROCEDURE NOTES: 

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
  2. Plastic (Part 5500), plastic-tipped or paraffin coated metal forceps must be used with any silver solution to prevent precipitation of silver salts. No metals of any kind should be in contact with any silver solution. Only glass thermometers should be used.
  3. Staining at higher temperatures means faster development but may cause precipitate to form in the working silver solution and deposit on slides. Maintaining silver solution between 45°C-60°C will minimize precipitate.
  4. Drain slides after each step to prevent solution carry over.
  5. Do not allow sections to dry out at any point during procedure.
  6. The suggested microwave procedure has been tested at Newcomer Supply. This procedure is a guideline and techniques should be developed for use in your laboratory.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 239-243.
  2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic”. American Journal of Clinical Pathology 25 (1955): 975-979.
  3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 3 (1981): 115-119.
  4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
  5. Modifications developed by Newcomer Supply Laboratory.

 

   

SOLUTION: 500 ml
Alpha Amylase 1%, Aqueous Part 1905B

 

Additionally Needed:

Periodic Acid Schiff (PAS) Glycogen Control Slides Part 4540
Periodic Acid 0.5%, Aqueous Part 13308
Schiff Reagent, McManus Part 1371
Hematoxylin Stain, Harris Part 12013
Acid Alcohol 1% Part 10011
Lithium Carbonate, Saturated Aqueous Part 12215
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Coplin Jar, Plastic Part 5184 (for glycogen digestion microwave modification)

 

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Alpha Amylase 1%, Aqueous is a convenient ready-to-use glycogen digestion solution for aiding in further identification of mucosubstances used in conjunction with the Periodic Acid Schiff (PAS) Stain procedure.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

 

STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Digestion Step: See Procedure Note #3.
    1. Two control slides and two patient slides are needed.
    2. Label one control slide and one patient slide “with”.
    3. Label the other control slide and patient slide “without”.
    4. Place slides labeled “without” in separate Coplin jar of distilled water; hold for Step #5.
    5. Apply Alpha Amylase 1%, Aqueous to slides labeled “with” for 30 minutes at room temperature.     
    6. Proceed to Step #5.
  4. Digestion Microwave Modification:  See Procedure Note #4.
    1. Follow Steps #3a through #3d.
    2. Place slides labeled “with” in a plastic Coplin jar containing Alpha Amylase 1%, Aqueous and microwave  for 1 minute at 37°C. Let sit in warm solution for an additional minute.
  5. Combine all slides for remaining steps; wash in running tap water for 1 minute, rinse in distilled water.
  6. Place in Periodic Acid 0.5%, Aqueous (Part 13308) for 10 minutes.
  7. Wash in three changes of tap water; rinse in distilled water.
  8. Place in Schiff Reagent, McManus (Part 1371) for 20 minutes.
  9. Wash in lukewarm tap water for 5 minutes.
  10. Stain with Hematoxylin Stain, Harris (Part 12013), 1-5 minutes, depending on preference of nuclear stain intensity.
  11. Wash in tap water for 2-3 minutes.
  12. Differentiate in Acid Alcohol 1% (Part 10011); 1-2 quick dips.
  13. Wash in tap water for 1 minute.
  1. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215); 3-4 dips.
  2. Wash in several changes of tap water; rinse in distilled water.
  3. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Glycogen Magenta
Glycogen digestion Absence of magenta
Acid & neutral epithelial mucin Magenta
Nuclei Blue

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Slides labeled “with” will be treated with amylase digestion, slides labeled “without” will not be treated for digestion.
  4. The suggested microwave procedure has been tested at Newcomer Supply.  This procedure is a guideline and techniques should be developed for use in your laboratory.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-171, 180.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
  3. Sheehan, Dezna C., and Barbara B.Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168.
  4. Modifications developed by Newcomer Supply Laboratory.

 

 

Technical Memo 1

Schiff Reagent, McManus for Periodic Acid Schiff (PAS) Stain 

 

 

SOLUTION:                                                                                                            

125 ml 250 ml 500 ml 1 Liter 4 Liters
Schiff Reagent, McManus Part 1371A Part 1371E Part 1371B Part 1371C Part 1371D

 

Additionally Needed For Periodic Acid Schiff (PAS) Stain:

Periodic Acid Schiff (PAS) Glycogen Control Slides Part 4540
Periodic Acid 0.5%, Aqueous Part 13308
Hematoxylin Stain, Harris Modified Part 1201
Acid Alcohol 1% Part 10011
Lithium Carbonate, Saturated Aqueous Part 12215
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

For storage requirements and expiration date refer to individual product labels.

APPLICATION:

Newcomer Supply Schiff Reagent, McManus a crucial element in the Periodic Acid Schiff (PAS) Stain and for staining glycoproteins, epithelial mucins, basement membranes and fungal cell walls. Schiff Reagent, McManus is used in a variety of staining procedures that include:

  • PAS with and without diastase
  • Alcian Blue/PAS for mucins
  • PAS/Light Green for Fungus
  • Feulgen Reaction for demonstration of DNA

Schiff Reagent, McManus is a stable and reliable product that is conveniently stored at room temperature for ready-to-use staining.   Although initially colorless, Schiff Reagent, McManus when in direct contact with skin, clothing, countertops, floors and other surfaces, will react and leave a bright magenta stain that is difficult to remove.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions manufactured by Newcomer Supply, Inc.

Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.

 

PAS STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Place in Periodic Acid 0.5%, Aqueous (Part 13308) for 10 minutes.
  4. Wash in three changes of tap water; rinse in distilled water.
  5. Place in Schiff Reagent, McManus for 20 minutes.
    1. See Procedure Notes #3, #4 and #5.
  6. Wash slides in lukewarm tap water for 5-10 minutes.
  7. Stain with Hematoxylin Stain, Harris Modified (Part 1201), 1-5 minutes, depending on preference of nuclear stain intensity.
  8. Wash in tap water for 2-3 minutes.
  9. Differentiate in Acid Alcohol 1% (Part 10011); 1-2 quick dips.
  10. Wash in tap water for 1 minute.
  11. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215); 3-4 dips.
  12. Wash in several changes of tap water; rinse in distilled water.
  13. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Glycogen Magenta
Acid & neutral epithelial mucin Magenta
Fungal cell walls Red to purple
Basement membranes Red to purple
Nuclei Blue

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Once used, do not pour Schiff Reagent, McManus back into the original bottle and/or mix with fresh solution.
  4. To test quality of Schiff Reagent, McManus reactivity;
    1. Add a few drops of Schiff Reagent, McManus to 10 ml of Formaldehyde 37-40%, ACS (Part 1089) or Formalin 10%, Phosphate Buffered (Part 1090).
    2. Good Schiff reagent will rapidly turn reddish-purple.
    3. Deteriorating Schiff reagent will have delayed reaction and turn deep blue-purple.
  5. All glassware/plasticware containing Schiff Reagent, McManus should be thoroughly rinsed with running tap water to remove residual stain prior to standard glassware cleaning.
  6. Digestion steps can be employed in the PAS procedure for further identification of mucosubstances.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 168-174, 180.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.137-141.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 164-168, 245.
  4. Modifications developed by Newcomer Supply Laboratory.

Technical Memo 2: 

Schiff Reagent, McManus for Fungus, PAS/Light Green 

SOLUTION:                                                                                                         

125 ml 250 ml 500 ml  1 Liter 4 Liters
Schiff Reagent, McManus Part 1371A Part 1371E Part 1371B Part 1371C Part 1371D

 

Additionally Needed For Fungus Stain, PAS/Light Green:

Fungus, PAS, Aspergillus sp., Artificial Control Slides

                         OR

Fungus, PAS, Candida sp., Artificial Control Slides

Part 4232

 

Part 4233

Periodic Acid 0.5%, Aqueous Part 13308
Light Green SF Yellowish Stain 0.1%, Aqueous Part 12203
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Schiff Reagent, McManus, a crucial element in the Fungus, PAS/Light Green procedure, is used for staining fungus and glycogen in tissue sections.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin section cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.

 

FUNGUS, PAS/LIGHT GREEN STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Place in Periodic Acid 0.5%, Aqueous (Part 13308) for 5 minutes.
  4. Wash in three changes of tap water; rinse in distilled water.
  5. Drain slides of  excess water and stain in Schiff Reagent, McManus for 20 minutes.
  6. Wash gently in lukewarm tap water for 10 minutes to allow pink color to develop.
  7. Counterstain in Light Green SF Yellowish Stain 0.1%, Aqueous (Part 12203) for 5 seconds.
    1. See Procedure Note #3.
  8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Fungal cell walls and glycogen Red to magenta
Background Pale green

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Increase or decrease staining time in Light Green SF Yellowish Stain 0.1%, Aqueous for preference of counterstain intensity.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 321-323.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245.
  3. Modifications developed by Newcomer Supply Laboratory.

 

 

  

KIT INCLUDES:                                                                     

Part 9155A    
Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic 250 ml
Solution B: Lithium Carbonate 0.5%, Aqueous 250 ml
Solution C: Cresyl Violet Stain, Aqueous 250 ml
Solution D: Acetic Acid 10%, Aqueous 50 ml

 

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed are two complimentary unstained positive control slides for initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers.

 

Additionally Needed:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Alcohol, Ethyl Denatured, 70% Part 10844
Coplin Jar, Plastic Part 5184 (for microwave modification)

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Luxol Fast Blue (LFB) – Cresyl Violet Stain Kit, with included microwave modification, is for the demonstration of myelin and Nissl substance in central nervous system and peripheral nerve tissues.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090) 

Technique: Paraffin sections cut at 8-10 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Prepare Working Lithium Carbonate 0.05%; combine and mix well;
    1. Solution B: Lithium Carbonate 0.5%, Aqueous 10 ml
    2. Distilled Water                90 ml
    3. Use 50 ml in each of Steps #7 & #10a.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.
    1. Stop at 95% ethyl alcohol; no distilled water rinse.
    2. See Procedure Notes #1 and #2.
  2. Incubate slides in Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic for 2 hours at 60°C or overnight at 37°C; cover tightly.

        Microwave Modification: See Procedure Note #3.

    1. Place slides in a plastic Coplin jar containing Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic and microwave at 70°C for 10 minutes.
  1. Rinse slides quickly in 95% ethyl alcohol, 2-3 dips.
  2. Rinse slides in distilled water.
  3. Differentiate each slide individually in Working Lithium Carbonate 0.05% (Step #2) for 10-15 seconds with agitation until gray matter and white matter are colorless and in high contrast with stained tissue.
  4. Further differentiate in 70% ethyl alcohol, until gray and white matter can be distinguished. Do not over differentiate.
  5. Rinse slides in distilled water.
  6. Complete differentiation:
    1. One dip in Lithium Carbonate 0.05%, Aqueous (Step #2).
    2. Dip in two changes of 70% ethyl alcohol until green/blue white matter sharply contrasts with colorless gray matter.
  7. Rinse thoroughly in distilled water.
  8. Prepare Cresyl Violet Working Solution:
    1. Solution C: Cresyl Violet Stain, Aqueous            40 ml
    2. Solution D: Acetic Acid 10%, Aqueous 7 drops
    3. Combine, mix well and filter.
    4. Directly before use, heat filtered solution to 57°C in microwave; hold in oven.
  9. Stain in heated Cresyl Violet Working Solution; 6 minutes in oven.
  10. Rinse in distilled water.
  11. Dehydrate quickly to maintain Cresyl Violet Stain in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Myelin Blue
Nissl substance and nuclei Violet
Neurons Pink to violet

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. The suggested microwave procedure has been tested at Newcomer Supply. This procedure is a guideline and techniques should be developed for use in your laboratory.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 214-215.
  2. Klüver, Heinrich, and Elizabeth Barrera. “A Method for the Combined Staining of Cells and Fibers in the Nervous System.” Journal of Neuropathology and Experimental Neurology4 (1953): 400-403.
  3. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 494-495.
  4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining ALK-Lymphoma and negative staining breast adenocarcinoma.

Fixation: Formalin 10%, Phosphate Buffered.

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Storage: 2-8°C in a light deprived and humidity controlled environment.

Intended Use:  To verify histological techniques and reagent reactivity.

 

PRODUCT DESCRIPTION:

The enclosed cell line control slides are developed from a process that allows production of compact cell preparations, cultured from human cell lines, that retain their cellular morphology and are tissue-like in composition.  These cell lines are standardized and manufactured to provide consistent results from slide to slide.  Each slide contains two, 2mm diameter cell line cores which demonstrate positive and negative expression for the specific marker.

 

 

 

 

APPLICATION:

Newcomer Supply ALK-Lymphoma (anaplastic lymphoma kinase) Cell Line Control Slides are for the positive immunohistochemical staining of ALK-Lymphoma with a NPM (nucleophosmin) gene translocation, associated with anaplastic large cell lymphoma (ALCL).

Validate positive ALK-Lymphoma Cell Line Control Slides per your laboratories standard operating procedure.

 

Convenient slide holder made of heavy duty plastic. Tray designed to hold slide in vertical position for easy left to right reading. Slides rest at a slightly upward angle. Designed for easy tipping of slide to pick up. Stackable trays so that slide will not touch tray above. Holds 20 slides (3″ x 1″). Dimensions: 8 1/8″ x 11 3/4″ x 11/16″ H). Available in 4 colors.

 

  

HELICOBACTER STAIN KIT INCLUDES:      

Part 9130A
Solution A: Periodic Acid 1%, Aqueous 250 ml
Solution B: Sodium Metabisulfite 5%, Acidified 250 ml
Solution C: Alcian Yellow Stain 1%, Alcoholic 250 ml
Solution D: Toluidine Blue Stock Stain 1%, Aqueous   10 ml
Solution E: Sodium Hydroxide 3%, Aqueous   10 ml

 

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed with this kit are two complimentary unstained positive control slides to be used for the initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and control slides may be available for purchase under separate part numbers.

 

Additionally Needed:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Acetic Acid 3%, Aqueous Part 10017 (for more intense Alcian Yellow staining)

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Helicobacter, Toluidine Blue/Alcian Yellow Stain Kit procedure is used for screening and detection of Helicobacter pylori in gastrointestinal biopsy specimens.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1.   See Procedure Note #1.
  2. Oxidize in Solution A: Periodic Acid 1%, Aqueous for 10 minutes.
  3. Wash slides well with tap water.
  4. Treat slides with Solution B: Sodium Metabisulfite 5%, Acidified for 5 minutes.
  5. Wash in tap water for 5 minutes.
    1.   See Procedure Note #2.
  6. Stain in Solution C: Alcian Yellow Stain 1%, Alcoholic for 5 minutes.
  7. Wash well in tap water.
  8. Prepare fresh Toluidine Blue Stain Working Solution; combine and mix well.
    1.  Distilled Water                                                              50 ml
    2.   Solution D: Toluidine Blue Stock Stain 1%, Aqueous  0.5 ml
    3.   Solution E: Sodium Hydroxide 3%, Aqueous               2 drops
  9. Stain in fresh Toluidine Blue Stain Working Solution for 3-5 minutes depending on preference of stain intensity.
  10. Wash well in tap water.
  11. Allow slides to air-dry completely.
  12. Clear dried slides in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
    1. See Procedure Note #3.

 

RESULTS:

Helicobacter pylori   Blue
Mucin Yellow
Background Pale blue

 

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. For a more intense Alcian Yellow stain, add a step of Acetic Acid 3%, Aqueous (Part 10017) for 1-3 minutes prior to Step #6.  Do not rinse slides and proceed directly into Solution C: Alcian Yellow Stain 1%, Alcoholic.
  3. The elimination of the dehydration steps is necessary to retain organism staining.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 225-226.
  2. Leung, Jennifer, Kevin Gibbon and Robert Vartaniam. “Rapid Staining Method for Helicobacter pylori in Gastric Biopsies.” The Journal of Histotechnology 19.12 (1996): 131-132.
  3. Modifications developed by Newcomer Supply Laboratory.

 

  

ALCIAN BLUE/PAS STAIN KIT INCLUDES: 

Part 91022A Part 91022B
Solution A: Acetic Acid 3%, Aqueous 250 ml 500 ml
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250 ml 500 ml
Solution C: Periodic Acid 0.5%, Aqueous 250 ml 500 ml
Solution D: Schiff Reagent, McManus 250 ml 500 ml
Solution E: Hematoxylin Stain, Mayer Modified 250 ml 500 ml

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed with this kit are two complimentary unstained positive control slides to be used for the initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers .

Additionally Needed:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Alcian Blue/PAS Stain Kit procedure is used to differentiate between acidic epithelial mucins (sialomucin, sulfomucin) and neutral epithelial mucin and is a means of detecting the overall presence of mucins.  Acidic mucins are stained with the Alcian Blue technique while neutral mucins and glycogen are stained by the PAS reaction.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  2. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
  3. Place slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous for 15 minutes.
  4. Wash slides in gently running tap water 1-2 minutes; rinse in distilled water.
  5. Place in Solution C: Periodic Acid 0.5%, Aqueous for 5 minutes.
  6. Wash in running tap water for 1-2 minutes; rinse in distilled water.
  7. Place in Solution D: Schiff Reagent, McManus for 10 minutes.
  8. Wash in lukewarm tap water for 5-10 minutes.
  9. Stain lightly in Solution E: Hematoxylin Stain, Mayer Modified for 1 minute.
  10. Rinse in running tap water for 1-2 minutes.
  11. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucin Violet
Neutral epithelial mucin Magenta
Glycogen Magenta
Stromal (mesenchymal) mucin Blue
Nuclei Pale blue

 

PROCEDURE NOTES:

  1. Drain staining rack/slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during staining procedure.
  3. Newcomer Supply Schiff Reagent, McManus is stored at room temperature.  There is no benefit to store this product at 4°C.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 173-174.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 150-151
  3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Alcian Blue/PAS quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Alcian Blue/PAS Stain Kit:  Part 91022A/B Individual Stain Solution
Solution A: Acetic Acid 3%, Aqueous 250/500 ml Part 10017
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250/500 ml Part 1003
Solution C: Periodic Acid 0.5%, Aqueous 250/500 ml Part 13308
Solution D: Schiff Reagent, McManus 250/500 ml Part 1371
Solution E: Hematoxylin Stain, Mayer Modified 250/500 ml Part 1202

 

APPLICATION:

Newcomer Supply Alcian Blue/PAS Control Slides are for the positive histochemical staining and differentiation of acidic epithelial mucins (sialomucin, sulfomucin), stromal (mesenchymal) mucin, neutral mucins and glycogen.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
  4. Place slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous for 15 minutes.
  5. Wash slides in gently running tap water for 1-2 minutes; rinse in distilled water.
  6. Place in Solution C: Periodic Acid 0.5%, Aqueous for 5 minutes.
  7. Wash in running tap water for 1-2 minutes; rinse in distilled water.
  8. Place in Solution D: Schiff Reagent, McManus for 10 minutes.
  9. Wash in lukewarm tap water for 5-10 minutes.
  10. Stain lightly in Solution E: Hematoxylin Stain, Mayer Modified for 1 minute.
  11. Rinse in running tap water for 1-2 minutes.
  12. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Violet
Neutral epithelial mucin Magenta
Glycogen Magenta
Stromal (mesenchymal) mucin Blue
Nuclei Pale blue

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 173-174.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 150-151
  3. Modifications developed by Newcomer Supply Laboratory.

SOLUTION:  500 ml 1 Liter
Tartrazine Stain 1.5%, Aqueous Part 14015A Part 14015B

 

Additionally Needed:

Gram, Multi-Tissue, Artificial Control Slides

                           OR

Gram+ & Gram- Bacteria, Artificial Control Slides

Part 4256

     OR

Part 4255

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Crystal Violet Stain 1%, Aqueous, Brown-Hopps Part 1041
Iodine, Gram, Aqueous Part 1140
Acetone, ACS Part 10014
Basic Fuchsin Stain 0.25%, Aqueous Part 1011
Gallego Solution Part 1098
Acetone-Xylene 1:1 Part 10015

For storage requirements and expiration date refer to individual product labels.

APPLICATION:

Newcomer Supply Brown-Hopps Modified Gram Stain with Tartrazine is a modification of the original Gram stain technique. Tartrazine, a synthetic water soluble, lemon yellow colored azo dye, provides a safe alternative to picric acid stains. Tartrazine Stain 1.5%, Aqueous provides yellow background staining, replacing the picric acid-acetone counterstain and the disposable issues associated with picric acid.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. If necessary, heat dry tissue sections/slides in oven
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Stain in Crystal Violet Stain 1%, Aqueous, Brown-Hopps (Part 1041) for 2 minutes.
  4. Rinse well in distilled water.
  5. Mordant in Iodine, Gram, Aqueous (Part 1140) for 5 minutes.
  6. Rinse well in distilled water.
  7. Blot excess water from slide; decolorize one slide at a time  in Acetone (Part 10014) until the blue stops running, 1-2 dips.
    1. Sections should be very light gray in color.
  8. Quickly rinse in running tap water.
  9. Place in Basic Fuchsin Stain 0.25% Aqueous (Part 1011) for 5 minutes.
  10. Rinse well in running tap water.
  11. Differentiate sections in Gallego Solution (Part 1098) for 5 minutes.
  12. Rinse in running tap water. Blot water off slide(s) but not to dryness.
    1. Proceed with Steps #13 to #16 one slide at a time.
  13. Place in Tartrazine Stain 1.5%, Aqueous for 1 minute.
  14. Rinse well in running tap water.
  15. Dip in Acetone, 1-2 quick dips.
  16. Dip in Acetone-Xylene 1:1 (Part 10015) for 5 dips.
  17. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Gram-positive bacteria Blue
Gram-negative bacteria Red
Nuclei Red
Background tissue Yellow

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Brown, Robert C., and Howard C. Hopps. “Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.” American Journal of Clinical Pathology 60.2 (1973): 234-240.
  2. Dapson, Janet Crookham, and Richard Dapson. Hazardous Materials in the Histopathology Laboratory: Regulations, Risks, Handling, and Disposal. 4th ed. Battle Creek, MI: Anatech, 2005. 150, 182, 266.
  3. Mercado, Gene. “Modifying the Modification: How to Redden Shy Gram Negatives.” The Journal of Histotechnology 25.2 (2002): 115-116.
  4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 235.
  5. Modifications developed by Newcomer Supply Laboratory.

Slotted lid and base for max fluid flow. Large labeling areas on 3 sides. Anterior edge @ 45° angle for manual or instrument labeling. Covers detached and packaged separately in the case.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal organ and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Spirochete quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Spirochete, Animal Control Slides are for the positive immunohistochemical staining of spirochetes, the causative agent of a variety of diseases such as; syphilis, bejel, pinta, yaws and lyme.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Treponema pallidum primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply HRP-Polymer solution. Incubate for 20 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  14. Rinse slides in four changes of distilled water.
  15. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  16. Rinse slides in warm tap water to blue sections.
  17. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Spirochete positive expression Brown
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare Treponema pallidum (polyclonal ACA135) is the concentrated primary antibody used.  Dilute primary antibody to 1/50 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Biocare Mach 2™ Universal HRP-Polymer Detection (M2U522) is the HRP-Polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare Treponema pallidum Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Biocare Mach 2™ Universal HRP-Polymer Detection datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

 

The Power of CaviCide in a Wipe!

Pre-saturated with CaviCide, CaviWipes Surface Disinfection Towelettes are a cleaner and disinfectant in one.  CaviWipes Surface Disinfection Towelettes are durable,non-woven, non-abrasive towelettes that offer quick, easy-to-use, time-saving convenience.  They are recomended  for use on non-porous surfaces and fixtures.  CaviWipes Surface Disinfection Towelettes should not be disposed of in the toilet.

CaviWipes Surface Disinfection Towelettes are intended for use in health care settings such as:

  • hospitals
  • laboratories
  • clinics
  • dental offices
  • opthalmic offices and
  • veterinary facilities

 

Features of the CaviWipes Surface Disinfection Towelettes:

  • Prevents cross-contamination of infected surfaces while maintaining material compatibility
  • Low Alcohol (17.20% Isopropanol)
  • Effective against Staphylococcus aureus with reduced susceptibility to vancomycin and influenza A2 Virus
  • Kills TB in 3 minutes and MRSA, HIV-1 and HBV and HCV in 2 minutes
  • Cleaner and disinfectant in one
  • Stays fully saturated – better fluid capacity means the wipes won’t dry out
  • Convenient, ready-to-use towel and CaviCide in one
  • Nonabrasive – for use on non-porous surfaces
  • Superior surface contact – durable, nonwoven towels won’t bunch up during use
  • Packaged in flat packs – minimizing storage space
  • Resealable packaging

 

CaviWipes Surface Disinfection Towelettes are effective against the following microorganisms on hard, non-porous surfaces when used as directed:

Kill Time
Mycobacterium bovis BCG 3 minutes
Pseudomonas aeruginosa 3 minutes
Salmonella enterica 3 minutes
Staphylococcus aureus 3 minutes
Trichophyton mentagrophytes 3 minutes
Methicillin Resistant Staphylococcus aureus (MRSA) 2 minutes
Staphylococcus aureus with Reduced Susceptibility to Vancomycin 2 minutes
Vancomycin Resistant Enterococcus faecalis (VRE) 2 minutes
Hapatitis B Virus (HBV) 2 minutes
Hepatitis C Virus (HCV) 2 minutes
Herpes Simplex Virus Type 1 2 minutes
Herpes Simplex Virus Type 2 2 minutes
Human Immunodeficiency Virus (HIV-1) 2 minutes
Influenza A2 Virus 2 minutes

 

 

The Polyscience 28 Liter Digital Water Bath offers excellent temperature control, range and uniformity.

 

Temperature Range and Dimensions of the Polyscience 28 Liter Digital Water Bath:

Temperature Range: Ambient +5ºC to 100ºC

Working Access: (L x W x D) 9.5″ x 17″ x 8″

Overall Dimensions: (L x W x H) 17.5″ x 22.5″ x 18″

 

Features of the Polyscience 28 Liter Digital Water Bath:

  • Hinged gable cover
  • Digital Temp. Controller with 3 pre-sets for frequently used temps.
  • Calibration capability
  • Local lockout to help prevent unauthorized set-point changes
  • Reservoir drain
  • Primary & automated safety thermostats
  • ETL Certification

 

 

The Polyscience 20 Liter Digital Water Bath offers excellent temperature control, range and uniformity.

 

Temperature Range and Dimensions of the Polyscience 20 Liter Digital Water Bath:

Temperature Range: Ambient +5ºC to 100ºC

Working Access: (L x W x D) 9.5″ x 17″ x 6″

Overall Dimensions: (L x W x H) 22.5″ x 17.5″ x 15.5″

 

Features of the Polyscience 20 Liter Digital Water Bath:

  • Hinged gable cover
  • Digital Temp. Controller with 3 pre-sets for frequently used temps.
  • Calibration capability
  • Local lockout to help prevent unauthorized set-point changes
  • Reservoir drain
  • Primary & automated safety thermostats
  • ETL Certification

 

 

The Polyscience 10 Liter Digital Water Bath offers excellent temperature control, range and uniformity.

 

Temperature Range and Dimensions of the Polyscience 10 Liter Digital Water Bath:

Temperature Range: Ambient +5ºC to 100ºC

Working Access: (L x W x D) 10.6″ x 11.6″ x 6″

Overall Dimensions: (L x W x H) 17″ x 15.5″ x 15.5″

 

Features of the Polyscience 10 Liter Digital Water Bath:

  • Hinged gable cover
  • Digital Temp. Controller with 3 pre-sets for frequently used temps.
  • Calibration capability
  • Local lockout to help prevent unauthorized set-point changes
  • Reservoir drain
  • Primary & automated safety thermostats
  • ETL Certification

 

 

The Polyscience 5 Liter Digital Water Bath offers excellent temperature control, range and uniformity.

 

 

Temperature Range and Dimensions of the Polyscience 5 Liter Digital Water Bath:

 

Temperature Range: Ambient +5ºC to 100ºC

Working Access: (L x W x D) 5″ x 10.8″ x 6″

Overall Dimensions: (L x W x H) 12″ x 14.5″ x 13.5″

 

 

Features of the Polyscience 5 Liter Digital Water Bath:

 

  • Hinged gable cover
  • Digital Temp. Controller with 3 pre-sets for frequently used temps.
  • Calibration capability
  • Local lockout to help prevent unauthorized set-point changes
  • Primary & automated safety thermostats
  • ETL Certification

 

 

The PolyScience 2 Liter Digital Water Bath offers excellent temperature control, range and uniformity.

 

Temperature Range/Dimensions of the PolyScience 2 Liter Digital Water Bath:

Temperature Range: Ambient +5ºC to +100ºC

Working Access: (L x W x D) 3.9″ x 4.3″ x 6″

Overall Dimensions: (L x W x H) 12″ x 9″ x 13.5″

 

Features of the PolyScience 2 Liter Digital Water Bath:

  • Hinged gable cover
  • Digital Temp. Controller with 3 pre-sets for frequently used temps.
  • Calibration capability
  • Local lockout to help prevent unauthorized set-point changes
  • Primary & automated safety thermostats
  • ETL Certification

 

  

KIT INCLUDES:                                                                                             

Part 9172A Part 9172B
Solution A: Zinc Formalin Sensitizer 250 ml 500 ml
Solution B: Silver Nitrate 1%, Aqueous 250 ml 500 ml
Solution C: Gum Mastic 2.5%, Alcoholic 175 ml x 2 350 ml x 2
Ingredient D: Hydroquinone, Powder 5 Grams 10 Grams
Mini Sampling Spoon

 

COMPLIMENTARY POSITIVE CONTROL SLIDES: Enclosed with this kit are two complimentary unstained positive control slides to be used for the initial verification of staining techniques and reagents.  Verification must be documented by running one Newcomer Supply complimentary positive control slide along with your current positive control slide for the first run. Retain the second complimentary control slide for further troubleshooting, if needed.

Individual stain solutions and additional control slides may be available for purchase under separate part numbers.

Additionally Needed:

Hydrochloric Acid 5%, Aqueous Part 12086 (for acid cleaning glassware)
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Acidulated Water pH 4.0-4.1 Part 10013
Coplin Jar, Plastic Part 5184 (for microwave modification)

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Steiner-Chapman Modified Silver Stain Kit procedure, with included microwave modification, is used for staining spirochetes, Helicobacter pylori, Legionella pneumophila, other nonfilamentous bacteria and fungus. This modified method eliminates the use of Uranyl Nitrate and its regulatory and disposal requirements.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

  • Mercury fixatives will inhibit silver staining

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. All glassware/plasticware must be acid cleaned prior to use.
    1. See Procedure Notes #1 and #2.
  2. Preheat Solution A: Zinc Formalin Sensitizer to 60°C.
    1. Do not preheat if using Microwave Modification.
  3. Preheat Solution B: Silver Nitrate 1%, Aqueous to 60°C.
    1. Do not preheat if using Microwave Modification.
  4. Prepare Hydroquinone Working Solution; combine and mix well.
    1. Ingredient D: Hydroquinone, Powder      0.5 gm (or one rounded scoop with reusable mini sampling spoon)
  1. Distilled water                                          25 ml 
  2. Save for use in Step #13. 

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with Acidulated Water pH 4.0-4.1 (Part 10013); two changes, 5 minutes each.
    1. See Procedure Notes #3 and #4.
  2. Sensitize slides in preheated Solution A: Zinc Formalin Sensitizer (Step #2) in a 60°C water bath or oven for 15 minutes.

        Microwave Modification:  See Procedure Note #5.

  1. Place slides in a plastic Coplin jar containing Solution A: Zinc Formalin Sensitizer and microwave to 60°C.  
  2. Remove from microwave; incubate and agitate in warm solution under fume hood for an additional 5 minutes.
  3. See Procedure Note #6.
  1. Rinse in several changes of Acidulated Water pH 4.0-4.1 (Part 10013).
  2. Place slides in preheated Solution B: Silver Nitrate 1%, Aqueous (Step #3) and incubate in a 60°C water bath or oven for 1 hour.

Microwave Modification: 

  1. Place sides in a plastic Coplin jar containing Solution B: Silver Nitrate 1%, Aqueous and microwave to 60°C.  
  2. Remove from microwave and let stand an additional 1 minute 30 seconds; agitate occasionally for even heat distribution.
  1. Rinse well in several changes of distilled water.
    1. Excessive rinsing may cause nuclei to pick up silver.
  2. Dip briefly in two changes each of 95% and 100% ethyl alcohols.
  3. Place slides in Solution C: Gum Mastic 2.5%, Alcoholic for 5 minutes.
  4. Air-dry slides for 1-5 minutes until slides are milky white.
  5. Prepare fresh Reducing Solution by combining:
    1. Solution C: Gum Mastic 2.5 %, Alcoholic           15 ml
    2. Hydroquinone Working Solution (Step #4)         25 ml
    3. Filter, then add and mix well:
    4. Solution B: Silver Nitrate 1%, Aqueous               0.3 ml
  6. Rinse air-dried slides quickly in distilled water.
  7. Place slides in fresh Reducing Solution. Incubate in a 45°C water bath for 10-30 minutes with frequent agitation; examine microscopically at 10 minutes.
    1. Check staining progress at timed intervals.
    2. Tissue will turn tan in color; bacteria will be black when the tissue reaches a golden brown color.
    3. Dip in warm distilled water before and after examination.

        Microwave Modification:  See Procedure Note #7.

  1. Heat slides in a plastic Coplin jar containing fresh Reducing Solution and microwave at 45°C for 30 seconds.  
  2. Remove from microwave; continue to incubate in warm solution for an additional 2 minutes.
  1. Wash for 3 minutes in running tap water; rinse in distilled water.
  2. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Spirochetes Dark brown to black
Helicobacter pylori Dark brown to black
Legionella pneumophila Dark brown to black
Nonfilamentous bacteria and fungus Dark brown to black
Background Golden brown

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.  Cleaning glassware with bleach is not equivalent to acid washing.
  2. Plastic (Part 5500), plastic-tipped or paraffin coated metal forceps must be used with any silver solution to prevent precipitation of silver salts.  No metals of any kind should be in contact with any silver solution. Only glass thermometers should be used.
  3. Drain staining rack/slides after each step to prevent solution carry over.
  4. The use of Acidulated Water pH 4.0-4.1 rinses (Steps #5 and #7) is recommended to ensure proper tissue pH, enhanced staining and reduced background staining.
  5. The suggested microwave procedure has been tested at Newcomer Supply using an “EB Sciences”, 850 watt microwave oven with temperature probe and agitation tubes.  This procedure is reproducible in our laboratory.  It is nonetheless a guideline and techniques should be developed for your laboratory which meet the requirements of your situation. Microwave devices should be placed in a fume hood or vented into a fume hood, according to manufacturer’s instructions, to prevent exposure to chemical vapors.
  6. Heating Zinc Formalin Sensitizer will result in liberation of toxic fumes. Use appropriate protective actions.
  7. The Reducing Solution contains alcohol and will reduce its boiling point. Do not boil solution. Adjust microwave times and power levels accordingly.
  8. The use of some xylene substitutes may result in diminished spirochete staining.  If using a xylene substitute exercise caution and closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

REFERENCES:

  1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 238-239.
  2. Chapman, Clifford, and Lorelei Margeson. “Use of Zinc Formalin as a Sensitizer in Silver Stains for Spirochetes.” The Journal of Histotechnology 19.2 (1996): 135-138.
  3. Steiner, Gabriel, and Grete Steiner. “New Simple Silver Stain for Demonstration of Bacteria, Spirochetes and Fungi in Sections of Paraffin Embedded Tissue Blocks.” Journal of Laboratory Clinical Medicine 29 (1944). 868-871.
  4. Modifications developed by Newcomer Supply Laboratory.

SOLUTIONS: 125 ml 500 ml 1 Liter 4 Liters
Hydrochloric Acid 20%, Aqueous Part 12087A Part 12087B
Schiff Reagent, McManus Part 1371A Part 1371B Part 1371C Part 1371D

Additionally Needed:

Normal Tonsil Custom Tissue Slides Part CT39790A
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Light Green SF Yellowish Stain 0.2%, Aqueous Part 12202

 

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

The Newcomer Supply Feulgen Reaction procedure is for the demonstration of DNA (deoxyribonucleic acid) in tissue sections.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

  1. See Procedure Note #1.

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.

 

STAINING PROCEDURE:

  1. Prepare Hydrochloric Acid Working Solution; combine and mix well.
    1. Hydrochloric Acid, 20% Aqueous            16 ml
    2. Distilled Water                                          24 ml
    3. Preheat and maintain Hydrochloric Acid Working Solution at 60°C prior to use. 
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #2 and #3.
  3. Hydrolyze sections in Hydrochloric Acid Working Solution at 60°C for 10 minutes.
    1. See Procedure Notes #4 and #5.
  4. Place slides directly in Schiff Reagent, McManus for 45 minutes.
    1. See Procedure Note #6.
  5. Wash in running tap water for 5 minutes; rinse in distilled water.
  6. Counterstain in Light Green SF Yellowish Stain 0.2%, Aqueous (Part 12202) for 1 minute.
  7. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

DNA Red-purple
Nuclei Red-purple
Background Green

 

PROCEDURE NOTES:

  1. Bouin fixed tissue is unsatisfactory for use with Feulgen reaction.
  2. Drain staining rack/slides after each step to prevent solution carry over.
  3. Do not allow sections to dry out at any point during staining procedure.
  4. For optimal results it is critical to preheat and maintain Hydrochloric Acid Working Solution at 60°C during the hydrolysis process.
  5. Prolonged exposure to hydrochloric acid may over-hydrolyze sections with poor staining results.
  6. Sodium Metabisulfite 5%, Acidified Aqueous (Part 13855) rinses can be added directly after Schiff Reagent, McManus to enhance Schiff staining.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008.224-225.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-Instructional Text. 4th ed. Chicago: ASCP Press, 2015. 126-127.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 150.
  4. Modifications developed by Newcomer Supply Laboratory.

 

  

FAT STAIN KIT INCLUDES:
Part 9119A
Solution A: Propylene Glycol 100%, ACS 250 ml
Solution B: Oil Red O Stain, Propylene Glycol 250 ml
Solution C: Propylene Glycol 85%, Aqueous 250 ml
Solution D: Hematoxylin Stain, Mayer Modified 250 ml

Individual stain solutions and additional control slides may be available for purchase under separate part numbers .

Additionally Needed:

Formalin 10%, Phosphate Buffered Part 1090
Lithium Carbonate, Saturated Aqueous

              OR

Scott Tap Water Substitute

Part 12215

 

Part 1380

Mount-Quick Aqueous Part 6271A

 

For storage requirements and expiration date refer to individual bottle labels.

 

FAT STAIN KIT APPLICATION:

Newcomer Supply Fat, Oil Red O, Propylene Glycol Stain Kit procedure is classified as a physical staining method and is used for identification of fat/lipid in frozen sections.

 

FAT STAIN KIT METHOD:

Fixation:   Fresh tissue or formalin fixed unprocessed tissue (See Procedure Note #1)

Technique:   Frozen sections cut at 8-10 microns mounted on adhesive slides

Solutions:     All solutions manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Kits are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the kit may contain extra volumes.

 

FAT STAIN KIT PROCEDURE:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
  1. See Procedure Note #2.
  1. Rinse sections carefully in two changes of distilled water.
  2. Blot off excess water and place slides in Solution A: Propylene Glycol 100%, ACS for 2-5 minutes.
  3. Place slides directly into Solution B: Oil Red O Stain, Propylene Glycol for 1 hour.  Agitate occasionally or place Coplin jar on rotator/shaker with continuous gentle agitation.
  1. See Procedure Notes #3 and #4.
  1. Differentiate in Solution C: Propylene Glycol 85%, Aqueous with agitation for a minimum of 3 minutes.
  2. Rinse gently in two changes of distilled water.
  3. Counterstain with Solution D: Hematoxylin Stain, Mayer Modified, for 2-3 minutes, depending on preference of nuclear stain intensity.
  4. Wash gently in several changes of tap water.
  5. Blue slides in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
  1. See Procedure Note #5.
  1. Wash gently in several changes of tap water.
  2. Blot excess water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) mounting medium.
  1. See Procedure Note #6. 

RESULTS:

Fat Bright red
Nuclei Blue to dark blue

 

FAT STAIN KIT PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue post-fixation: Place specimen in tissue cassette, wash in running water for 5 minutes. Remove tissue from cassette and blot well removing all excess water from tissue. Freeze tissue (fresh or formalin fixed) according to your laboratory protocol.
  2. Frozen formalin fixed tissue does not require an additional formalin fixation step.
  3. To decrease staining time; preheat Solution B: Oil Red O Stain, Propylene Glycol in a 60°C oven and decrease incubation time to 7-10 minutes.
  4. If a filmy precipitate develops in Solution B: Oil Red O Stain, Propylene Glycol filter with coarse filter paper.
  5. The use of a bluing agent in this procedure is an optional step.
  6. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles under coverslip; soak slide in warm water until coverslip can easily be removed. Blot excess water from slide and remount with new coverslip and Mount-Quick Aqueous mounting medium.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 184-186.
  2. Prophet, Edna B., Bob Mills, Jacquelyn Arrington, and Leslie Sobin. Laboratory Methods in Histotechnology. Washington, D.C.: American Registry of Pathology. 1992.178.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 205.
  4. Modifications developed by Newcomer Supply Laboratory.

Safety Overview – Click Here

SOLUTION:                                                  

500 ml  1 Liter
Van Gieson Stain Part 1404A Part 1404B

 

Additionally Needed:

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Van Gieson Stain is a connective tissue stain and/or counterstain that combines acid fuchsin and saturated picric acid, with acid fuchsin selectively staining collagen and osteoid tissue red and the picric acid component staining muscle, elastin, fibrin and cytoplasm yellow.

This stain solution is commonly used in elastic stains, referred to as the Verhoeff-Van Gieson (VVG) technique. Other procedures that use Van Gieson Stain include;

  • Bile Stain, Hall’s Method
  • Colloidal Iron, MÏ‹ller-Mowry Stain
  • Sulfated Alcian Blue (SAB) Stain
  • Van Gieson’s Picric Acid-Fuchsin Stain

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique: Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  2. Proceed with selected stain procedure:
    1. Verhoeff-Van Gieson (VVG) Elastic Stain
    2. Bile Stain, Hall’s Method
    3. Colloidal Iron, MÏ‹ller-Mowry Stain
    4. Sulfated Alcian Blue (SAB) Stain
    5. Van Gieson’s Picric Acid-Fuchsin Stain
    6. Or other appropriate stain procedure
  3. Counterstain in Van Gieson Stain for 3 to 5 minutes.
    1. See Procedure Note #3.
  4. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Collagen Red
Muscle, elastin, fibrin, cytoplasm Yellow
Other tissue components Dependent on stain procedure used

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. The picric acid element may act as a decolorizer in some procedures.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 146-147.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 166-167.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 189, 196.
  4. Modifications developed by Newcomer Supply Laboratory.

 

  

SET INCLUDES:                                                                                       

Part 14034A Part 14034B
Solution A: Neutral Red Stain 1%, Alcoholic 250 ml 500 ml
Solution B: Fast Green Stain 1%, Alcoholic 100 ml 200 ml

 

Additionally Needed For Gram Stain, Hucker-Twort:                                                                                       

Gram, Multi-Tissue, Artificial Control Slides

                            OR

Gram+ & Gram- Bacteria, Artificial Control Slides

Part 4256

    OR

Part 4255

Crystal Violet-Oxalate Stain, Alcoholic, Hucker-Twort Part 10422
Iodine, Weigert & Lugol, Aqueous  Part 12092
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Acetone, ACS Part 10014

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Twort’s Gram Stain Set provides stain solutions for the Gram Stain, Hucker-Twort, a rapid and simple procedure that stains gram-positive and gram-negative bacteria without the use of picric acid.  The Neutral Red Stain combined with a Fast Green counterstain provides clear detection of any red gram-negative bacteria present against a green background.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Sets are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure. Some solutions in the set may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Filter Crystal Violet-Oxalate Stain, Alcoholic, Hucker-Twort (Part 10422) with high quality filter paper.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  2. Stain in freshly filtered Crystal Violet-Oxalate Stain, Alcoholic, Hucker-Twort (Step #2) for 30 seconds.
  3. Rinse quickly in distilled water.
  4. Mordant in Iodine, Weigert & Lugol, Aqueous (Part 12092) for 20 seconds.
  5. Rinse quickly in distilled water.
  6. Decolorize individually with Acetone, ACS (Part 10014); 2 quick dips.
    1. Or until majority of purple stain is removed and tissue remains light gray.
  7. Rinse quickly in distilled water.
  8. Prepare fresh Twort Stain; combine and mix well.
    1. Solution A: Neutral Red Stain 1%, Alcoholic      9 ml
    2. Solution B: Fast Green Stain 1%,  Alcoholic      3 ml
    3. Distilled Water                                                 30 ml
    4. Use within 30 minutes.
  9. Stain in fresh Twort Stain for 2 minutes.
  1. Rinse quickly in distilled water and carefully blot dry.
  2. Agitate slides quickly in clean Acetone, ACS to dehydrate (do not use any alcohols).
    1. See Procedure Notes #2 and #3.
  3. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:                        

Gram-positive bacteria Dark blue
Gram-negative bacteria Red
Cytoplasm and red blood cells Shades of green
Nuclei Red

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. If needed, add extra dips in acetone to further differentiate and dehydrate sections.
    1. Check microscopically to avoid over-differentiation.
  3. The use of alcohol will remove Neutral Red staining.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bancroft, John D., and Alan Stevens. Theory and Practice of Histological Techniques. 3rd ed. Edinburgh: Churchill Livingstone, 1990. 290-292.
  2. Culling, C.F.A. Handbook of Histopathological and Histochemical Techniques. 3rd ed. London: Butterworth, 1974. 393-395.
  3. Twort, F.W., “An Improved Neutral Red, Light Green Double Staining for Animal Parasites, Microorganisms and Tissues”. Journal of State Medicine 32. (1924). 351.
  4. Modifications developed by Newcomer Supply Laboratory.

Toluidine Blue Stain for Mast Cells – Technical Memo

 

SOLUTION:                                                                                                                                      

250 ml 500 ml 1 Gallon
Toluidine Blue Stain 0.1%, Aqueous Part 14027A Part 14027B Part 14027D

 

Additionally Needed:

Mast Cell Control Slides

                OR

Mast Cell, Animal Control Slides

Part 4410

     OR

Part 4412

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Toluidine Blue Stain for Mast Cells is for the demonstration of mast cells, characterized as cells filled with basophilic granules, associated with inflammation and allergic reactions, which stain metachromatically with toluidine blue.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  2. Place slides in Toluidine Blue Stain 0.1%, Aqueous for 10 minutes.
  3. Rinse well in distilled water.
  4. Dehydrate quickly through two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
    1. See Procedure Note #3.

 

RESULTS:

Mast cells Deep blue-violet
Background Blue

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Metachromasia of mast cell granules is stable and staining will be maintained during dehydration steps.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Broome, Michelle and Beth Villarreal. “Differential Staining of Mast Cells with Toluidine Blue”. The Journal of Histotechnology 35.1 (2012): 27-30.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.188.
  3. Modifications developed by Newcomer Supply Laboratory.

 

 

Toluidine Blue Stain for Mohs Technique – Technical Memo

 

SOLUTION:

250 ml 500 ml 1 Gallon
Toluidine Blue Stain 0.1%, Aqueous Part 14027A Part 14027B Part 14027D

 

Additionally Needed:

Alcohol, Ethyl Denatured, 70% Part 10844
Alcohol, Ethyl Denatured, 95% Part 10842
Alcohol, Ethyl Denatured, 100% Part 10841
Xylene, ACS Part 1445

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Toluidine Blue Stain for Mohs Technique provides a rapid staining method for Mohs micrographic surgery (MMS), useful when evaluating frozen skin samples for basal cell carcinoma (BCC).  Toluidine Blue imparts an identifiable staining pattern if BCC is present that will highlight islands of blue staining basal cell carcinoma and metachromatically stain surrounding mucopolysaccharides/stroma pink.

 

METHOD:

Fixation: 70% Ethyl Alcohol (Part 10844)

Technique:  Frozen sections cut at 5-7 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Fix tissue sections in 70% Ethyl Alcohol for 30-60 seconds.
    1. See Procedure Note #1.
  2. Wash well in distilled water.
  3. Stain slides in Toluidine Blue Stain 0.1%, Aqueous for 30-60 seconds, depending on preference of stain intensity.
  4. Wash gently in distilled water.
  5. Dehydrate quickly through one change of 95% ethyl alcohol; 1 quick dip and then two changes 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
    1. See Procedure Note #2.

 

RESULTS:

Islands of basal cell carcinoma Deep blue to purple
Surrounding mucopolysaccharides/stroma Pink to magenta
Background Blue
Nuclei Dark blue

 

PROCEDURE NOTES:

  1. Section thickness and fixation timing may affect staining quality.
  2. Alcohol will work as a differentiator. Proceed quickly through dehydration steps to maintain Toluidine Blue stain.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendation for clearing step.

 

REFERENCES:

  1. Arnon, Ofer, Ronald Rapini, Adam Mamelak, and Leonard Goldberg. “Mohs Micrographic Surgery: Current Techniques.” IMAJ 12 (2010): 431-35.
  2. Gross, Kenneth G. Mohs Surgery: Fundamentals and Techniques. St. Louis: Mosby, 1999. 125-138.
  3. Modifications developed by Newcomer Supply Laboratory.

 

PRACTICAL APPLICATION OF THIAZINE STAIN SOLUTION:

Histology:

  • Stain for touch preps on fresh tissue during intraoperative procedures (See Reference 1)
  • Rapid, inexpensive stain to identify H. pylori in gastrointestinal tissue sections (See Reference 2)

 

Cytology:

  • Adequacy assesment of FNA’s (See Reference 3 & 4)

 

Hematology:

 

REFERENCES:

  1. Yulin Liu M.D., Jan F. Silverman M.D., Charles D. Sturgis M.D., Henry G. Brown M.D., David J. Dabbs M.D. and Stephen S. Raab M.D.  Utility of intraoperative consultation touch preparations, Diagnostic Cytopathology Volume 26, Issue 5, pages 329-333, May 2002.
  2. Skipper, Ray, DeStephano, Don B. A Rapid Stain for Campylobacter pylori in Gastrointestinal Tissue Sections Using Diff-Quik® J of Histotechnology Volume 12 Issue 4. 1989, pp. 303-304.
  3. Silverman JF, Frable WJ.  The use of the Diff-Quik stain in the immediate interpretation of fine-needle aspiration biopsies.  Diagon Cytopathology 1990; 6:366-369.
  4. Rosemary H. Tambouret, MD; Guliz A. Barkan, MD; Daniel F.I. Kurtycz, MD; Vijayalakshmi Padmanabhan, MD  Rapid on-site evaluation – how practice varies.  CAP Today Volume 128, No. 5, pages 29-31, May 2014

SOLUTIONS: 125 ml
Sudan IV Stain, Herxheimer Alcoholic Part 1400A

 

Additionally Needed:

Alcohol, Ethyl Denatured, 70% Part 10844
Hematoxylin Stain, Mayer Modified Part 1202
Lithium Carbonate, Saturated Aqueous

                 OR

Scott Tap Water Substitute

Part 12215

    OR

Part 1380

Mount-Quick Aqueous Mounting Medium Part 6271A
For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Sudan IV Stain, Herxheimer Alcoholic procedure is used for identification of fat/lipid in frozen sections. Herxheimer method refers to an acetone/alcohol solvent mixture; the acetone component of this solution may dissolve out small amounts of lipid.

Sudan dyes are a group of fat/lipid soluble solvent dyes, also known as lysochromes.  These solvent dyes readily stain fat/lipid due to the fact that the dyes are more soluble in lipid than in the solvents from which they are applied.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue

  1. See Procedure Note #1.

​Technique: Frozen sections cut at 8 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered for 1 minute.
    1. See Procedure Note #2.
  2. Rinse sections carefully in two changes of distilled water.
  3. Rinse in Alcohol, Ethyl Denatured, 70% (Part 10844).
  4. Stain in Sudan IV Stain, Herxheimer Alcoholic for 10 minutes.
    1. Keep tightly capped to avoid evaporation.
  5. Differentiate quickly in Alcohol, Ethyl Denatured, 70% to remove excess stain.
  6. Wash thoroughly in distilled water.
  7. Counterstain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2-3 minutes.
  8. Wash gently in several changes of tap water.
  9. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    1. The use of a bluing agent is optional.
  10. Wash gently in several changes of tap water.
  11. Blot excess water from slide; coverslip with Mount-Quick Aqueous Mounting Medium.
    1. See Procedure Note #3.

 

RESULTS:

Fat Orange/red
Nuclei Blue

 

PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue:
    1. Place specimen in tissue cassette; wash in running water for 5 minutes.
    2. Remove tissue from cassette; blot well, removing all excess water from tissue.
    3. Freeze tissue according to laboratory protocol.
  2. Frozen formalin fixed tissue does not require additional formalin fixation.
  3. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles or to recoverslip;
    1. Soak slide in warm water until coverslip is easily  removed.
    2.  Blot excess water from slide.
    3. Remount with new coverslip and Mount-Quick Aqueous Mounting Medium.

 

REFERENCES:

  1. Culling, C. F. A. Handbook of Histopathological and Histochemical Techniques: (including Museum Techniques). 3rd ed. London: Butterworth, 1974. 359-362.
  2. Kiernan, J. A. Histological and Histochemical Methods: Theory and Practice. 3rd ed. London, Ontario: Arnold, 2003. 251-254.
  3. Lillie, R. D., and Harold Fullmer. Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, 1976. 565-568.
  4. Modifications developed by Newcomer Supply Laboratory.

 

Tech Memo for Sudan III Stain, Saturated Alcoholic for Frozen Sections/For Fecal Fat:

 

SOLUTIONS: 500 ml
Sudan III Stain, Saturated Alcoholic Part 1390A

 

Additionally Needed:

Alcohol, Ethyl Denatured, 70% Part 10844
Hematoxylin Stain, Mayer Modified Part 1202
Lithium Carbonate, Saturated Aqueous

                   OR

Scott Tap Water Substitute

Part 12215

    OR

Part 1380

Mount-Quick Aqueous Mounting Medium Part 6271A

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Sudan III Stain, Saturated Alcoholic is used for identification of fat/lipid in frozen tissue sections. Sudan dyes are a group of fat/lipid soluble solvent dyes, also known as lysochromes.  These solvent dyes readily stain fat/lipid due to the fact that the dyes are more soluble in lipid than in the solvents from which they are applied.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue

  1. See Procedure Note #1.

​Technique: Frozen tissue sections cut at 8 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered for 1 minute.
    1. See Procedure Note #2.
  2. Rinse sections carefully in two changes of distilled water.
  3. Rinse in Alcohol, Ethyl Denatured, 70% (Part 10844).
  4. Stain in Sudan III Stain, Saturated Alcoholic for 10 minutes.
    1. Keep tightly capped to avoid evaporation.
  5. Differentiate in Alcohol, Ethyl Denatured, 70%.
  6. Wash thoroughly in distilled water.
  7. Counterstain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2-3 minutes.
  8. Wash gently in several changes of tap water.
  9. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    1. The use of a bluing agent is optional.
  10. Wash gently in several changes of tap water.
  11. Blot excess water from slide; coverslip with Mount-Quick Aqueous Mounting Medium.
    1. See Procedure Note #3.

 

RESULTS:

Fat Orange/red
Nuclei Blue

 

PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue post:
    1. Place specimen in tissue cassette, wash in running water for 5 minutes.
    2. Remove tissue from cassette; blot well, removing all excess water from tissue.
    3. Freeze tissue according to laboratory protocol.
  2. Frozen formalin fixed tissue does not require additional formalin fixation.
  3. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles or to recoverslip;
    1. Soak slide in warm water until coverslip is easily removed.
    2. Blot excess water from slide.
    3. Remount with new coverslip and Mount-Quick Aqueous Mounting Medium.

 

REFERENCES:

  1. Culling, C. F. A. Handbook of Histopathological and Histochemical Techniques: (including Museum Techniques). 3rd ed. London: Butterworth, 1974. 359-362.
  2. Kiernan, J. A. Histological and Histochemical Methods: Theory and Practice. 3rd ed. London, Ontario: Arnold, 2003. 251-254.
  3. Modifications developed by Newcomer Supply Laboratory.

 

Tech Memo for Sudan III Stain Alcoholic For Fecal Fat:

                                                             

SOLUTION:   500 ml
Sudan III Stain, Saturated Alcoholic  Part 1390A

 

Additionally Needed:

Alcohol, Ethyl Denatured, 95% Part 10842
Acetic Acid, Glacial, ACS Part 10010

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Sudan III Stain, Saturated Alcoholic is used for microscopic detection of increased fecal fat levels due to malabsorption.  This screening procedure for malabsorption and steatorrhea consists of two parts; the neutral fat stain and the split fat stain for fatty acids.

 

METHOD:

Fixation:  Fresh/unpreserved fecal material

  1. See Procedure Note #1.

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

 

STAINING PROCEDURE: METHOD FOR NEUTRAL FATS

  1. Examine within accepted time, a fresh fecal sample collected, stored and prepared per laboratory protocol.
    1. See Procedure Note #2.
  2. Place small aliquot of prepared fecal sample on a clean glass slide; approximately 5mm in diameter.
  3. Mix two drops of Alcohol, Ethyl Denatured, 95% (Part 10842) with the fecal sample on the slide.
  4. Add two drops of Sudan III Stain, Saturated Alcoholic to the fecal suspension on the slide; mix well.
  5. Coverslip and examine microscopically.

 

RESULTS:       

Neutral fats Red/orange refractile globules

 

STAINING PROCEDURE:  SPLIT FAT STAIN FOR FATTY ACIDS

  1. Prepare in advance 36% Acetic Acid, Glacial; mix well.
    1. Acetic Acid Glacial, ACS (Part 10010)          9 ml
    2. Distilled Water                                     16 ml
    3. Store at room temperature for up to 1 year.
  2. Examine within accepted time frame, a fresh fecal sample collected, stored and prepared per laboratory protocol.
    1. See Procedure Note #2.
  3. Place small aliquot of prepared fecal sample on a clean glass slide; approximately 5mm in diameter.
  4. Mix two drops of Alcohol, Ethyl Denatured, 95% (Part 10842) with the fecal sample on the slide.
  5. Add two drops of Sudan III Stain, Saturated Alcoholic to the fecal suspension on the slide; mix well.
  6. Add two drops of 36% Acetic Acid Glacial; mix well.
  7. Place on preheated hot plate (calibrated to 60°C or slightly higher) until bubbles appear.  Or hand hold slide over preheated hot plate until bubbles appear; quickly remove slide and reheat two additional times.
    1. See Procedure Note #3.
  8. Immediately coverslip and examine microscopically while warm.

 

RESULTS:      

Fatty acids Red/orange refractile globules

 

PROCEDURE NOTES:

  1. For optimum results fresh, unpreserved fecal material is required.
  2. Refer to laboratory protocols and guidelines for proper collection methods, specimen storage requirements and time frame that satisfactory testing results can be obtained.
  3. Sudan dyes dissolve in lipids at temperatures above the melting point of the lipid or when lipid is in liquid phase. The melting point of lipids in which fatty acid chains are saturated melt above 60°C. Most unsaturated lipids are liquid at room temperature (20°-22°C).

 

REFERENCES:

  1. Bauer, John D. Clinical Laboratory Methods. 9th ed. St. Louis: Mosby, 1982. 793-794.
  2. Fine, Kenneth D., and Frederick Ogunji. “A New Method of Quantitative Fecal Fat Microscopy and Its Correlation With Chemically Measured Fecal Fat Output.” American Journal of Clinical Pathology 113.4 (2000): 528-534.
  3. Fischbach, Frances, and Marshall Dunning. A Manual of Laboratory and Diagnostic Tests. 8th ed. Lippincott Williams & Wilkins, 2008. 303-304.
  4. Khouri, M.R., G. Huang, and Y.F. Shiau. “Sudan Stain of Fecal Fat: New Insight Into an Old Test.” Gastroenterology 96.2 (1989): 421-427.
  5. Kiernan, J. A. Histological and Histochemical Methods: Theory and Practice. 3rd ed. London, Ontario: Arnold, 2003. 251-252, 254.
  6. Modifications developed by Newcomer Supply Laboratory.

SOLUTIONS: 125 ml 250 ml
Sudan Black B Stain, Propylene Glycol Part 1401A Part 1401B

 

Additionally Needed:

Formalin 10%, Phosphate Buffered Part 1090
Propylene Glycol 100%, ACS Part 13391
Propylene Glycol 85%, Aqueous Part 133912
Nuclear Fast Red Stain, Kernechtrot Part 1255
Mount-Quick Aqueous Mounting Medium Part 6271A

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Sudan Black B Stain, Propylene Glycol procedure is used for identification of fat/lipid in frozen sections. Sudan black B is a lipid soluble solvent dye that readily stains neutral fats and phospholipids.

Sudan dyes are a group of fat/lipid soluble solvent dyes, also known as lysochromes.  These solvent dyes readily stain fat/lipid due to the fact that the dyes are more soluble in lipid than in the solvents from which they are applied.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue

  1. See Procedure Note #1.

​Technique: Frozen sections cut at 8-10 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
    1. See Procedure Note #2.
  2. Rinse sections carefully in two changes of distilled water.
  3. Blot off excess water and dehydrate slides in Propylene Glycol 100%, ACS (Part 13391) for 10-15 minutes.
  4. Place directly into Sudan Black B Stain, Propylene Glycol for 30 minutes to 1 hour.  Agitate occasionally or place Coplin jar on rotator/shaker with continuous gentle agitation.
    1. See Procedure Note #3.
  5. Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) for 3 minutes with agitation.
  6. Rinse gently in distilled water.
  7. Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
    1. Shake solution well before use; do not filter.
  8. Wash gently in several changes of tap water.
    1. See Procedure Note #4.
  9. Blot excess water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) Mounting Medium.
    1. See Procedure Note #5.

 

RESULTS:

Fat Blue-black
Nuclei Red

 

PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue:
    1. Place specimen in tissue cassette, wash in running tap water for 5 minutes.
    2. Remove tissue from cassette; blot well, removing all excess water from tissue.
    3. Freeze tissue according to laboratory protocol. 
  2. Frozen formalin fixed tissue does not require additional formalin fixation.
  3. To decrease staining time; preheat Sudan Black B Stain, Propylene Glycol in a 60oC oven; stain for 3-10 minutes.
  4. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in coverslipped sections.
  5. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles or to recoverslip;
    1. Soak slide(s) in warm water until coverslip is easily removed.
    2. Blot excess water from slide.
    3. Remount with new coverslip and Mount-Quick Aqueous Mounting Medium.

 

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 192-193.
  2. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 185-186.
  3. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 464-465.
  4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 204-205.
  5. Modifications developed by Newcomer Supply Laboratory.

 

 

SOLUTION:                                                                                               

500 ml
Safranin O Stain 0.25%, Aqueous Part 1360A

Additionally Needed: 

Crystal Violet Stain 1%, Aqueous, Brown-Hopps Part 1041
Iodine, Gram, Aqueous Part 1140
Acetone, ACS Part 10014

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Safranin O Stain 0.25%, Aqueous provides the preferred counterstain for Gram staining of microbiology smears.  The Gram Stain technique, is used for differential staining of gram-positive and gram-negative bacteria in both smears and tissue sections.

 

METHOD:

Technique:  Flat staining rack method.

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

 

STAINING PROCEDURE:

  1. Prepare within an accepted time frame, a well-made smear(s) per laboratory protocol, with a focus on uniform distribution of material.
  2. Allow slides to thoroughly air-dry before staining.
  3. Fix slides according to laboratory protocol.
    1. See Procedure Note #1. 
  4. Place slides on flat staining rack suspended over sink.
    1. See Procedure Note #2.
  5. Flood fixed smears with Crystal Violet Stain 1%, Aqueous, Brown-Hopps (Part 1041) for 45-60 seconds.
  6. Drain off Crystal Violet Stain and rinse well in distilled water.
  7. Mordant smears in Iodine, Gram, Aqueous (Part 1140) for 45-60 seconds.
  8. Rinse well in running tap water to remove excess iodine.
  9. Blot one slide at a time and individually decolorize in Acetone (Part 10014) until color stops running off the smear; 5-10 seconds.
    1. See Procedure Note #3.
  10. Quickly rinse in distilled water to remove excess acetone.
  11. Counterstain in Safranin O Stain 0.25%, Aqueous for 30-60 seconds.
    1. See Procedure Note #4.
  12. Wash well in distilled water.
  13. Allow slides to air-dry.
  14. Examine microscopically with oil immersion.

 

RESULTS:

Gram-positive bacteria Blue to blue/black
Gram-negative bacteria Pink to red
Background Yellow

 

PROCEDURE NOTES:

  1. Gentle heat or methanol are both accepted methods of smear fixation.
  2. Do not allow smears to dry out at any point during staining procedure.
  3. To decolorize slower, dip slides in Alcohol, Ethyl Denatured, 95% (Part 10842) in Coplin jar for 10-60 seconds or until color stops running off the smear.
  4. For enhanced counterstaining;
    1. Safranin O Stain 0.25%, Aqueous 10 ml
    2. 95% Ethyl Alcohol (Part 10842)               1 ml
    3. Combine; mix well and stain for 30-60 seconds.

 

REFERENCES:

  1. Kidd, Larry. “Histology vs. Microbiology – The Gram Stain the Easy Way.” The Journal of Histotechnology 7.2 (1984): 85-86.
  2. Lillie, R. D., and Harold Fullmer. Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, 1976. 726-727.
  3. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia:  Elsevier Saunders, 2011. 1080.
  4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 235.
  5. Modifications developed by Newcomer Supply Laboratory.

 

SOLUTION:                                                                                                    

250 ml 500 ml
Oil Red O Stain, Propylene Glycol Part 12772A Part 12772B

 

Additionally Needed:

Formalin 10%, Phosphate Buffered Part 1090
Propylene Glycol 100%, ACS Part 13391
Propylene Glycol 85%, Aqueous Part 133912
Hematoxylin Stain, Mayer Modified Part 1202
Lithium Carbonate, Saturated Aqueous

                   OR

Scott Tap Water Substitute

Part 12215

     OR

Part 1380

Mount-Quick Aqueous Part 6271A

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Oil Red O Stain, Propylene Glycol procedure is classified as a physical staining method and is used for identification of fat/lipid in frozen sections.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue

Technique: Frozen sections cut at 8-10 microns on adhesive slides

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

  1. Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
    1. See Procedure Note #2.
  2. Rinse sections carefully in two changes of distilled water.
  3. Blot excess water and place slides in Propylene Glycol 100%, ACS (Part 13391) for 2-5 minutes.
  4. Place directly into Oil Red O Stain, Propylene Glycol for 1 hour.  Agitate occasionally or place Coplin jar on rotator/shaker with continuous gentle agitation.
    1. See Procedure Notes #3 and #4.
  5. Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) with agitation for a minimum of 3 minutes.
  6. Rinse gently in two changes of distilled water.
  7. Counterstain with Hematoxylin Stain, Mayer Modified (Part 1202) for 2-3 minutes.
  8. Wash gently in several changes of tap water.
  9. Blue in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
    1. The use of a bluing agent is optional.
  10. Wash gently in several changes of tap water.
  11. Blot excess water from slide; coverslip with Mount-Quick Aqueous (6271A) mounting medium.
    1. See Procedure Note #5.

 

RESULTS:

Fat Bright red
Nuclei Blue to dark blue

 

PROCEDURE NOTES:

  1. To freeze formalin fixed unprocessed tissue:
    1. Place specimen in tissue cassette, wash in running tap water for 5 minutes.
    2. Remove tissue from cassette; blot well, removing all excess water from tissue.
    3. Freeze tissue according to laboratory protocol.
  2. Frozen formalin fixed tissue does not require additional formalin fixation.
  3. To decrease staining time; preheat Oil Red O Stain, Propylene Glycol in a 60°C oven; stain for 7-10 minutes.
  4. If a filmy precipitate develops in Oil Red O Stain, Propylene Glycol, filter with coarse filter paper.
  5. Use minimal pressure when applying coverslip or fat/lipid staining may be disturbed.  To remove trapped air bubbles or to recoverslip;
    1. Soak slide in warm water until coverslip is easily removed.
    2. Blot excess water from slide.
    3. Remount with new coverslip and Mount-Quick Aqueous mounting medium.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 184-186.
  2. Prophet, Edna B., Bob Mills, Jacquelyn Arrington, and Leslie Sobin. Laboratory Methods in Histotechnology. Washington, D.C.: American Registry of Pathology. 1992.178.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 205.
  4. Modifications developed by Newcomer Supply Laboratory.

  

SET INCLUDES:                                                                                                     

Part 1142A Part 1142B
Solution A: Silver Nitrate 500 ml 1000 ml
Solution B: Methenamine Borate 500 ml 1000 ml

 

Additionally Needed For Fungus Stain, Grocott Methenamine Silver, GMS:

Fungus, GMS, Multi-Tissue, Artificial Control Slides Part 4235
Hydrochloric Acid 5%, Aqueous Part 12086 (for acid cleaning glassware)
Chromic Acid 5%, Aqueous Part 10341
Sodium Bisulfite 1%, Aqueous Part 13821
Gold Chloride 0.1%, Aqueous Part 11285
Sodium Thiosulfate 2%, Aqueous Part 13888
Light Green SF Yellowish Stain 0.02%, Aqueous Part 12204
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842
Coplin Jar, Plastic Part 5184 (for microwave modification)

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Grocott Methenamine Silver Set, GMS with included microwave modifications, provides the silver staining solutions for the Fungus Stain, Grocott Methenamine Silver (GMS) procedure.  This is one of the best staining methods for demonstrating a variety of fungal organisms including: Pneumocystis carinii, Aspergillus, Blastomyces, Candida and Histoplasma.

When staining for Pneumocystis with other fungal organisms, running a separate control specific for Pneumocystis (Part 4556) is recommended.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 5 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply Stain Sets are designed to be used with Coplin jars filled to 40 ml following the staining procedure provided below.  Some solutions in the set may contain extra volumes.

 

PRESTAINING PREPARATION:

  1. All glassware/plasticware must be acid cleaned prior to use.
    1. See Procedure Notes #1 and #2.
  2. Prepare Silver-Methenamine Working Solution and mix well:
    1. Solution A: Silver Nitrate                        20 ml
    2. Solution B: Methenamine Borate            20 ml
  3. Preheat Silver-Methenamine Working Solution to 45°C – 60°C approximately 20-30 minutes before use.
    1. See Procedure Notes #3 and #4
    2. Save for use in Step #9.
    3. Do not preheat if using Microwave Modification; Step 10.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #5 and #6.
  2. Oxidize in Chromic Acid 5%, Aqueous (Part 10341) for 1 hour.

        Microwave Modification: See Procedure Note #7

  1. Oxidize slides in a plastic Coplin jar containing Chromic Acid 5%, Aqueous (Part 10341) and microwave for 1 minute and 20 seconds at 60°C.
  1. Wash well in running tap water; rinse in distilled water.
  2. Place in Sodium Bisulfite 1%, Aqueous (Part 13821) for 1 minute.
  3. Wash for 5 minutes in running tap water; rinse well in distilled water.
  4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #3) at 45°C-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
    1. Periodically remove control, rinse in warm distilled water, check microscopically for adequate silver impregnation.  Fungi should be dark brown. 
    2. If organisms are not sufficiently dark, return slides to warm silver solution.  Recheck at 2-3 minute intervals until desired intensity is achieved.
    3. Pneumocystis may take longer to stain than other fungus.
    4. Staining at room temperature will require longer incubation.
  5. Microwave Modification: See Procedure Note #7
    1. Incubate slides in a plastic Coplin jar containing Silver-Methenamine Working Solution (Step #2) and microwave for 1 minute at 70°C. 
    2. Check microscopically for adequate development.
    3. If additional incubation is required, return slides to warm Silver-Methenamine Working Solution.  Recheck at 2-3 minute intervals.
  6. Rinse in three to four changes of distilled water.
    1.   Do not use tap water at this step.
  7. Tone in Gold Chloride 0.1%, Aqueous (Part 11285) until sections turn gray; 20 seconds to 1 minute.
  8. Rinse well in distilled water.
  9. Remove unreduced silver in Sodium Thiosulfate 2%, Aqueous (Part 13888) for 2 minutes.
  10. Wash in running tap water for 5 minutes; rinse in distilled water.
  11. Counterstain in Light Green SF Yellowish Stain 0.02%, Aqueous (Part 12204) for 2 minutes.
  12. Dehydrate quickly in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium

 

RESULTS:

Fungi Crisp black cell walls with visible internal structures
Background Green
Mucin Taupe to dark gray

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.  Cleaning glassware with bleach is not equivalent to acid washing.
  2. Plastic (Part 5500), plastic-tipped, or paraffin coated metal forceps must be used with any silver solution to prevent precipitation of silver salts.  No metals of any kind should be in contact with any silver solution. Only glass thermometers should be used.
  3. Preheating Silver-Methenamine Working Solution to 45°C-60°C prior to incubation is suggested for timely silver development. A water bath can be used for preheating.  Begin preheating the silver solution approximately 20-30 minutes before use.
  4. Staining slides at higher temperatures will cause the development reaction to happen faster, but may also cause precipitate to form in the working silver solution and deposit on the slides.  Maintaining the silver solution between 45°C-60°C will help to minimize precipitate.
  5. Drain staining rack/slides after each step to prevent solution carry over.
  6. Do not allow sections to dry out at any point during staining procedure.
  7. The suggested microwave procedure has been tested at Newcomer Supply using an “EB Sciences”, 850 watt microwave oven with temperature probe and agitation tubes.  This procedure is reproducible in our laboratory.  It is nonetheless a guideline and techniques should be developed for your laboratory which meet the requirements of your situation. Microwave devices should be placed in a fume hood or vented into a fume hood, according to manufacturer’s instructions, to prevent exposure to chemical vapors.
  8. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 239-243.
  2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic”. American Journal of Clinical Pathology 25 (1955): 975-979.
  3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
  4. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
  5. Modifications developed by Newcomer Supply Laboratory.

 

 

CLICK HERE to see TRICHROME GOMORI, One-Step in Standard & Special Stains Section

SOLUTION: 500 ml 1 Liter
Giemsa Stock Stain Part 1120A Part 1120B

 

Additionally Needed:

Alcohol, Methanol Anhydrous, ACS Part 12236
Phosphate Buffer, pH 7.0 Part 1331

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Giemsa Stain is a simple one-step method designed to demonstrate differential staining of cells types in peripheral blood smears and bone marrow smears/films as well as a method for detecting rickettsia, bacteria and parasites.

 

METHOD:

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

STAINING PROCEDURE:

Prepare within an accepted time frame, a well-made blood smear or bone marrow smear/film per your laboratories protocol, with a focus on uniform cell distribution.

  1. Proceed with either the thin or thick smear/film staining method.

 

Thin Smear/Film Staining Method: See Procedure Notes #1 and #2.

  1. Allow smear to thoroughly air-dry prior to staining.
  2. Fix smear in Methanol: 1-2 minutes.
  3. Air-dry slides in a vertical position.
  4. Prepare fresh 1:20 Working Giemsa Stain; combine and mix well.
    1. Giemsa Stock Stain                           2 ml
    2. Phosphate Buffer, pH 7.0 (Part 1331)    40 ml
  5. Stain in Working Giemsa Stain for 20-30 minutes.
  6. Rinse briefly in Phosphate Buffer, pH 7.0 or distilled water.
  7. Air-dry slides in a vertical position.
  8. If coverslip is preferred, allow slides to air-dry; coverslip with compatible mounting medium.

 

Thick Smear/Film Staining Method: See Procedure Notes #1 and #2.

  1. Allow smear to thoroughly air-dry prior to staining; several hours or overnight.
  2. Proceed directly to stain; do not place in fixative.
  3. Prepare fresh 1:50 Working Giemsa Stain; combine and mix well.
    1. Giemsa Stock Stain                           1 ml
    2. Phosphate Buffer, pH 7.0 (Part 1331)    50 ml
  4. Stain in Working Giemsa Stain for 50 minutes.
  5. Rinse briefly in Phosphate Buffer, pH 7.0 or distilled water.
  6. Air-dry slides in a vertical position.
  7. If coverslip is preferred, allow slides to air-dry; coverslip with compatible mounting medium.

 

RESULTS:

Erythrocytes Orange – pink to rose
Platelets Red to purple granules with blue halo

Granulocytes

Neutrophils Nucleus – Dark blue to violet
Cytoplasm – Pink
Granules – Purple to lilac
Eosinophils Nucleus – Blue
Granules – Orange to pink
Basophils Nucleus – Deep blue to violet
Granules – Deep blue to violet

Mononuclear Cells

Lymphocytes Nuclei – Deep blue to violet
Cytoplasm – Light blue
Monocytes Nuclei – Light blue/purple
Cytoplasm – Pale gray/blue
Mast cells Nuclei – Deep blue to violet
Granules – Deep blue-violet
Malarial parasites Nucleus – Red chromatin dot
Cytoplasm – Blue
Rickettsia Bluish purple
Bacteria Blue

 

PROCEDURE NOTES:

  1. The timings provided are suggested ranges.  Optimal staining times will depend upon smear/film thickness and preference of stain intensity.
  2. Smears/films containing primarily normal cell populations require minimum staining time.
    1. Immature cells may require a longer staining time.
    2. Bone marrow smears/films may require a longer staining time.

 

REFERENCES:

  1. Bailey, W. Robert, and Elvyn Scott. Diagnostic Microbiology. 4th ed. St Louis: C. V. Mosby Company, 1974. 394.
  2. Garcia, Lynne Shore. Diagnostic Medical Parasitology. 5th ed. Washington DC: ASM Press, 2007. 888-889.
  3. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia:  Elsevier Saunders, 2011. 522-531.
  4. Modifications developed by Newcomer Supply Laboratory.

 

 

SOLUTIONS:                                             

250 ml 500 ml
Fouchet Reagent Part 1095A Part 1095B

 

Additionally Needed:

Bile Control Slides Part 4060
Van Gieson Stain Part 1404
Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Bile Stain, Hall’s Method is for the demonstration of bile (bilirubin) substances in tissue sections and to distinguish bile pigments from other tissue pigments.  The acidity of Fouchet Reagent works to oxidize bilirubin to biliverdin, resulting in a green color development with the Van Gieson Stain serving as a complimentary counterstain.

 

METHOD:

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Filter Fouchet Reagent with Grade 1 filter paper prior to use.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  2. Place slides in freshly filtered Fouchet Reagent for 5 minutes.
  3. Wash in three changes of tap water; rinse in distilled water.
  4. Stain sections in Van Gieson Stain (Part 1404) for 5 minutes.
  5. Rinse quickly in 95% ethyl alcohol.
  6. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Bile/bilirubin Emerald green to olive drab
Connective tissue Pink to red
Background Yellow

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 268-269.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 219.
  3. Modifications developed by Newcomer Supply Laboratory.

 

Technical Memo 1: Ziehl-Neelsen for AFB Stain

 

SOLUTION:                                                        

250 ml 500 ml 1 Liter
Carbol Fuchsin Stain, Ziehl-Neelsen Part 1030A Part 1030B Part 1030C

 

Additionally Needed for AFB Stain, Ziehl-Neelsen:

Acid Fast Bacteria (AFB) Control Slides Part 4011
Acid Alcohol 1% Part 10011
Light Green SF Yellowish Stain .1%, Aqueous

                          OR

Methylene Blue Stain 0.14%, Alcoholic 

Part 12203

     OR

Part 12401

Xylene, ACS Part 1445
Alcohol, Ethyl Denatured, 100% Part 10841
Alcohol, Ethyl Denatured, 95% Part 10842

 

For storage requirements and expiration date refer to individual bottle labels.

 

APPLICATION:

Newcomer Supply Carbol Fuchsin Stain, Ziehl-Neelsen, a crucial element in the AFB Stain, Ziehl-Neelsen is used to demonstrate the presence of acid-fast mycobacteria in tissue sections. Acid-fastness is a physical property of certain bacteria and cellular structures.  Carbol Fuchsin Stain, Ziehl-Neelsen, combines phenol and basic fuchsin that works to permeate the lipoid capsule of acid-fast organisms and renders them resistant to acid alcohol decolorization.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Filter Carbol Fuchsin Stain, Ziehl-Neelsen with filter paper whenever a thick sheen develops on solution surface.

 

STAINING PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  2. Stain in Carbol Fuchsin Stain, Ziehl-Neelsen for 15 minutes at room temperature. Keep solution covered.
    1. See Procedure Note #3.
  3. Rinse in running tap water for 2 to 3 minutes.
  4. Differentiate in Acid Alcohol 1% (Part 10011) until color no longer runs off the slide and sections are pale pink; 3 to 10 rapid dips.
  5. Wash in running tap water 3 to 5 minutes; rinse in distilled water.
  6. Counterstain with 3-6 dips in counterstain of choice;
    1. Light Green SF Yellowish Stain 0.1%, Aqueous (Part 12203)
    2. Methylene Blue Stain 0.14%, Alcoholic (Part 12401).  Do not overstain; sections should be pale blue.
  7. Rinse slides:
    1. Light Green SF Yellowish counterstain; rinse with one quick dip in distilled water or proceed directly to Step #10 without a distilled water rinse.
    2. Methylene Blue counterstain; rinse in running tap water for 1 minute; rinse in distilled water.
  8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid-fast bacilli  Bright red
Background Green (with Light Green SF Yellowish counterstain)
Background Pale blue (with Methylene Blue counterstain)

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. Sections can remain in Carbol Fuchsin Stain, Ziehl-Neelsen for up to 60 minutes without adverse effect. Additional differentiation may be required in Step #6.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 218-220.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
  3. Modifications developed by Newcomer Supply Laboratory.

Technical Memo 2: Ziehl-Neelsen for AFB, Fite Stain

 

SOLUTION:                                                                                                                    

250 ml 500 ml 1 Liter
Carbol Fuchsin Stain, Ziehl-Neelsen Part 1030A Part 1030B Part 1030C

 

Additionally Needed for AFB Stain, Fite:

Fite Stain, Nocardia Sp. Control Slides

                 OR

Abnormal Animal Spleen Custom Tissue Slides

Part 4215

    OR

Part CT28730A

Xylene/Peanut Oil, 2:1  Part 1449
Acid Alcohol 1% Part 10011
Light Green SF Yellowish Stain 0.1%, Aqueous

        OR

Methylene Blue Stain 0.5%, Aqueous

Part 12203

   OR

Part 12402

Xylene, ACS Part 1445

For storage requirements and expiration date refer to individual product labels.

 

APPLICATION:

Newcomer Supply Carbol Fuchsin Stain, Ziehl-Neelsen, a crucial element in the AFB Stain, Fite is used to detect the presence of either Nocardia sp. or Mycobacterium leprae sp. (causative agent of leprosy) in tissue sections.  Minor procedural variations are included for detection of either organism.

 

METHOD:

Fixation:  Formalin 10%, Phosphate Buffered (Part 1090)

Technique:  Paraffin Sections cut at 4 microns

Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

All Newcomer Supply stain procedures are designed to be used with Coplin jars filled to 40 ml following the provided staining procedure.

 

PRESTAINING PREPARATION:

  1. If necessary, heat dry tissue sections/slides in oven.
  2. Filter Carbol Fuchsin Stain, Ziehl-Neelsen with high quality filter paper.
  3. If staining for Nocardia sp., prepare Diluted Acid Alcohol Solution:
    1. Acid Alcohol 1% (Part 10011)                 20 ml
    2. Distilled water                                  20 ml                      

 

STAINING PROCEDURE:

  1. Deparaffinize slides in Xylene/Peanut Oil, 2:1 (Part 1449), two changes for 10  minutes each.
    1. See Procedure Note #1
  2. Drain slides, wipe off excess oil, and blot to opacity taking care to remove residual oil.
    1. See Procedure Note #2. 
  3. Stain in freshly filtered Carbol Fuchsin Stain, Ziehl-Neelsen for 15 minutes at room temperature.
  4. Rinse well in distilled water.
  5. Differentiation:
    1. For Nocardia sp.: Differentiate slides individually in Diluted Acid Alcohol Solution (Step #3) until background is pale pink; 10-20 dips.  Quickly rinse in distilled water and check microscopically for correct differentiation.
    2. For Mycobacterium leprae sp.: Differentiate slides individually in Acid Alcohol 1% (Part 10011) until sections are light pink; 5-10 dips.
  6. Rinse well in distilled water.
  7. Counterstain with 5-10 dips in counterstain of choice;
    1. Light Green SF Yellowish Stain 0.1%, Aqueous (Part 12203).
    2. Methylene Blue Stain 0.5%, Aqueous (Part 12402).  Do not overstain; sections should be pale blue.
  1. Rinse slides:
    1. Light Green SF Yellowish counterstain; rinse in distilled water.
    2. Methylene Blue counterstain; wash in running tap water, rinse in distilled.
  2. Blot excess water from slide and air-dry or oven-dry completely.
  3. Dip dried slides in xylene and coverslip with compatible mounting medium.

 

RESULTS:

Acid-fast bacilli and Mycobacterium leprae sp. Red
Nocardia sp. Red
Background Green (with Light Green SF Yellowish counterstain)
Background Pale blue (with Methylene Blue counterstain)

 

PROCEDURE NOTES:

  1. Acid-fastness of leprosy organisms is enhanced when the waxy capsule is protected by the mixture of xylene-peanut oil and avoidance of dehydrating solutions.
  2. It is important to blot well; residual oil may produce staining artifact.
  3. A small percentage of Nocardia sp. organisms may resist taking the red stain and remain green (or blue, depending upon counterstain used) due to growth phase of the individual organism.
  4. If using a xylene substitute, closely follow the manufacturer’s recommendations for coverslipping step.

 

REFERENCES:

  1. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 220-221.
  2. Fite, George, P.J. Cambre and M.H. Turner. “Procedure for Demonstrating Lepra Bacilli in Paraffin Sections”. Archives of Pathology 43 (1947). 624-625.
  3. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
  4. Modifications developed by Newcomer Supply Laboratory.

 

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining thyroid and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  PAX8 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply PAX8 (paired-box gene 8) Control Slides are for the positive immunohistochemical staining of PAX8, expressed in thyroid, renal cell, endometroid and ovarian carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply PAX8 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

PAX8 positive expression Brown nuclear staining
Myometrium Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque PAX8 (EP298) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque PAX8 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  PAX5 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply PAX5 (paired-box gene 5) Control Slides are for the positive immunohistochemical staining of PAX5, a transcription factor expressed throughout B-cell maturation and in most B-cell malignancies such as Hodgkin’s lymphoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply PAX5 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

PAX5 positive expression Brown nuclear staining
Adipose Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque PAX-5 (SP34) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque PAX-5 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining spleen.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Factor XIIIa quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Factor XIIIa (fibrin stabilizing factor) Control Slides are for the positive immunohistochemical staining of Factor XIIIa, a blood coagulation enzyme that crosslinks fibrin.  Factor XIIIa is used as a marker for fibro-histiocytic proliferations, dermal dendritic cells and to differentiate dermatofibroma from dermatofibrosarcoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Factor XIIIa primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Factor XIIIa positive expression Brown cytoplasmic staining 

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Factor XIIIa (EP3372) is the concentrated primary antibody used.  Dilute primary antibody to 1/300 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Factor XIIIa Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining kidney and negative staining thyroid.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Epithelial Membrane Antigen quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Epithelial Membrane Antigen (EMA) Control Slides are for the positive immunohistochemical staining of EMA, a marker of epithelial cells expressed in a variety of normal and neoplastic epithelia, including mammary and squamous epithelium and sweat glands. EMA is also noted to be expressed in a number of malignant lymphomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply EMA primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

EMA positive expression Brown cytoplasmic & membrane staining
Thyroid Negative
Nuclei  Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque EMA (E29) is the concentrated primary antibody used.  Dilute primary antibody to a 1/1200 dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque EMA Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  BCL2 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply BCL2 (B-cell lymphoma 2) Control Slides are for the positive immunohistochemical staining of BCL2. Frequently used to distinguish positive staining follicular lymphoma from negative reacting follicular hyperplasia.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply BCL2 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

BCL2 positive expression Brown cytoplasmic staining
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque BCL2 (124) is the concentrated primary antibody used.  Dilute primary antibody to 1/25 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque BCL2 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Gram positive staining rat lung, gram negative staining rat lung and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Brown-Hopps quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Gram, Brown-Hopps, Stain Kit: Part 9124A Individual Stain Solution
Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps 250 ml Part 1041
Solution B: Iodine, Gram, Aqueous 250 ml Part 1140
Solution C: Basic Fuchsin Stain 0.25%, Aqueous 250 ml Part 1011
Solution D: Gallego Solution 250 ml Part 1098
Solution E: Picric Acid-Acetone 0.05% 250 ml Part 13351
Solution F: Acetone-Xylene 1:1 250 ml Part 10015
Acetone, ACS Part 10014

 

APPLICATION:

Newcomer Supply Gram, Multi-Tissue, Artificial Control Slides are for the positive histochemical staining of gram positive and gram negative bacteria in separate tissue sections. Escherichia coli and Staphylococcus aureus purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Stain in Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps for 2 minutes.
  4. Rinse well in distilled water, ensuring excess stain is removed.
  5. Mordant in Solution B: Iodine, Gram, Aqueous for 5 minutes.
    1. Sections should turn black.
  6. Rinse well in running tap water, ensuring excess iodine is removed.
  7. Blot excess water from slide; decolorize one slide at a time in Acetone, ACS (Part 10014) until blue color stops running; 1-2 dips.
    1. Sections should be very light gray in color.
  8. Quickly rinse in running tap water to remove excess Acetone.
  9. Stain in Solution C: Basic Fuchsin Stain 0.25%, Aqueous; 5 minutes.
  10. Rinse well in running tap water.
  11. Differentiate sections in Solution D: Gallego Solution for 5 minutes.
  12. Rinse in running tap water.  Blot water off slide(s), but not to dryness.
    1. Proceed with Steps #13 to #16 one slide at a time.
  13. Dip quickly in Acetone, ACS (Part 10014); 1-2 dips.
  14. Dip directly in Solution E: Picric Acid-Acetone 0.05%; 3-10 dips.
  15. Dip quickly in Solution F: Acetone-Xylene 1:1; 5 dips.
  16. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Gram negative bacteria Red
Gram positive bacteria Blue/violet
Nuclei Red
Background tissue Yellow
Nonreactive lung Negative for gram positive/negative bacteria

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Brown, Robert C., and Howard C. Hopps. “Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.” American Journal of Clinical Pathology 60.2 (1973): 234-240.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 222-224.
  3. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Brown-Hopps quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

CONTROL SLIDE VALIDATION:

With Gram, Brown-Hopps Stain Kit: Part 9124A Individual Stain Solution
Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps 250 ml Part 1041
Solution B: Iodine, Gram, Aqueous 250 ml Part 1140
Solution C: Basic Fuchsin Stain 0.25%, Aqueous 250 ml Part 1011
Solution D: Gallego Solution 250 ml Part 1098
Solution E: Picric Acid-Acetone 0.05% 250 ml Part 13351
Solution F: Acetone-Xylene 1:1 250 ml Part 10015
Acetone, ACS Part 10014

 

APPLICATION:

Newcomer Supply Gram Positive & Gram Negative Bacteria, Artificial Control Slides are for the positive histochemical staining of gram positive and gram negative bacteria in the same tissue section.  Escherichia coli and Staphylococcus aureus purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #1 and #2.
  3. Stain in Solution A: Crystal Violet Stain 1%, Aqueous, Brown-Hopps for 2 minutes.
  4. Rinse well in distilled water, ensuring excess stain is removed.
  5. Mordant in Solution B: Iodine, Gram, Aqueous for 5 minutes.
    1. Sections should turn black.
  6. Rinse well in running tap water, ensuring excess iodine is removed.
  7. Blot excess water from slide; decolorize one slide at a time in Acetone, ACS (Part 10014) until blue color stops running; 1-2 dips.
    1. Sections should be very light gray in color.
  8. Quickly rinse in running tap water to remove excess Acetone.
  9. Stain in Solution C: Basic Fuchsin Stain 0.25%, Aqueous; 5 minutes.
  10. Rinse well in running tap water.
  11. Differentiate in Solution D: Gallego Solution for 5 minutes.
  12. Rinse in running tap water.  Blot water off slide(s), but not to dryness.
    1. Proceed with Steps #13 to #16 one slide at a time.
  13. Dip quickly in Acetone, ACS (Part 10014); 1-2 dips.
  14. Dip directly in Solution E: Picric Acid-Acetone 0.05%; 3-10 dips.
  15. Dip quickly in Solution F: Acetone-Xylene 1:1; 5 dips.
  16. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Gram positive bacteria Blue/violet
Gram negative bacteria Red
Nuclei Red
Background tissue Yellow
Nonreactive lung Negative for gram positive/negative bacteria

 

PROCEDURE NOTES:

  1. Drain slides after each step to prevent solution carry over.
  2. Do not allow sections to dry out at any point during procedure.
  3. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Brown, Robert C., and Howard C. Hopps. “Staining of Bacteria in Tissue Sections: A Reliable Gram Stain Method.” American Journal of Clinical Pathology 60.2 (1973): 234-240.
  2. Carson, Freida L., and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 222-224.
  3. Modifications developed by Newcomer Supply Laboratory.

 

 

Block Filing Cabinets

  • Six segmented, impact resistant plastic drawers with ID area per unit
  • Seven segmented rows per drawer.  Each row includes a foam block to support blocks in upright position.
  • Stackable cabinets have locking features to each other
  • Interfaces with other popular plastic block storage systems!
  • One piece construction with six drawers per unit
  • Identification writing area on each door
  • Each 6 drawer unit accommodates up to 1,500 blocks or 1,000 rings
  • Dimensions: 17 3/8″L x 9 1/2″W x 15 1/4″H

 

Block filing cabinet drawer

 

Locking pegs at each corner for stacking multiple units on one another

 

Filing cabinets stack securely together

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal organ.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Gomori Prussian Blue quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

CONTROL SLIDE VALIDATION:

With Iron, Gomori Prussian Blue Stain Kit: Part 9136A/B Individual Stain Solution
Solution A: Hydrochloric Acid 20%, Aqueous 125/250 ml Part 12087
Solution B: Potassium Ferrocyanide 10%, Aqueous 125/250 ml Part 13392
Solution C: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

 

APPLICATION:

Newcomer Supply Iron, Animal Control Slides are for the positive histochemical staining of ferric iron deposits in tissue sections.

 

PRESTAINING PREPARATION:

  1. Heat dry sections in oven according to your laboratory protocol
  2. To avoid the possibility of residual background iron staining, acid clean glassware is recommended in the staining procedure.
    1. See Procedure Note #1.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Notes #2 and #3.
  2. Prepare fresh Ferrocyanide Working Solution directly before use; combine and mix well.
    1. Solution A: Hydrochloric Acid 20%, Aqueous               20 ml
    2. Solution B: Potassium Ferrocyanide 10%, Aqueous    20 ml
  3. Place slides in fresh Ferrocyanide Working Solution for 20 minutes.
  4. Rinse in three changes of tap water; rinse in distilled water.
  5. Place in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
    1. Shake solution well before use; do not filter.
  6. Rinse well in distilled water.
    1. See Procedure Note #4.
  7. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Ferric iron deposits Bright blue
Nuclei Red
Cytoplasm Pink

 

PROCEDURE NOTES:

  1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
  2. Drain slides after each step to prevent solution carry over.
  3. Do not allow sections to dry out at any point during procedure.
  4. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
  5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 179-184.
  2. Sheehan, Dezna C., and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 217-218.
  3. Modifications developed by Newcomer Supply Laboratory.

 

The Activate™ Bleach Sprayer Dilution System dispenses a 10% bleach solution. The Activate™ Bleach Sprayer Dilution System has been made to meter and dispense 5.25% Sodium Hypochlorite NaOCL at a 10% (9:1) solution. 

The Activate™ Bleach Sprayer Dilution System has been widely recommended by CDC and other nationally recognized authorities

Features and Benefits of the Activate™ Bleach Sprayer Dilution System:

  • Automatically mixes water and bleach to make a fresh 10% solution every time the trigger is pulled.

  • Prevents bleach from degrading (ALWAYS AT FULL STRENGTH).

  • Elliminates safety hazard of mixing bleach by hand!

  • Fast contact time for practical disinfecting.

  • Sealed system prevents closed cap concerns.

  • Non-aerosol coarse spray.

  • Easy to install disposable bleach cartridges make approximately one gallon useable solution.

  • Sustainable packaging reduces plastic waste and saves in freight costs.

  • Ergonomic trigger sprayer is comfortable for high use.

 

Activate™ Bleach Sprayer Dilution System Practical Contact Times:

Viricide 30 Seconds (HIV-1 & HBV)
Bactericide 2 Minutes (MRSA & Bacteria)
Sporicide 4 Minutes (C.difficile Spores)

 

Specifications of the Activate™ Bleach Sprayer Dilution System:

Active Ingredient: 5.25% Sodium Hypochlorite
Dispensed Dilution Rate: 10% (1:9 bleach to water ratio)
Dispensed Available Chlorine: 5000 ppm
EPA Reg. No.: 75266-1
Personal Protection Dispensed Solution: Gloves, eye & clothing protection
Ventilation Requirement Dispensed Solution: Moderate ventilation required
Solubility: 100% in water
Odor Dispensed Solution: Light chlorine
pH Dispensed Solution: 9.5
Fire Hazard: None
Explosion Hazard: None
Storage Temperature: 50°-72°F
Shelf life: 12 Mos from mfg (when stored at 50°-72°F)
Incompatibilities: Ammonia, acids, reducing agents, heavy metals such as cobalt, copper, iron and nickel

 

How to Use the Activate Bleach Sprayer Dilution System

Step 1: Fill the bleach sprayer's empty water bottle with cool tap water, and then insert the sprayer's tube down into the filled water bottle. Close the bleach sprayer tab to lock the water bottle into place on the sprayer. 

 

Step 2:  Remove the threaded plastic cap from the bleach sprayer cartridge. Do not puncture or tamper with the plug that is in the neck of the bleach sprayer cartridge. It must remain intact in order for the bleach sprayer to dilute the bleach at the proper 10% dilution rate. 

 

Step 3:  Lock the bleach sprayer's cartridge onto the other side of the sprayer. 

 

Step 4:  Pointing the bleach sprayer away from you into a sink or basin, pump the trigger 20 times to prime the system. 

Step 5: The bleach sprayer is now ready to use. Hold the sprayer 6" to 8" from the surface, and spray thoroughly until the surface is completely wet. 

 

Step 6:  Refill the water bottle when empty. 

 

Step 7:  Always re-prime the bleach sprayer when installing a new bleach cartridge. Note that a safety feature ensures the bleach sprayer will not function if either the bleach sprayer's cartridge or the water bottle is empty. Periodically test the sprayed bleach solution for available chlorine with a high-level chlorine test strip. Always store your bleach system and additional bleach cartridges at room temperature (50º – 72º). 

 

exclamation,red,warning,alert,wrong,error  Note:  The bleach sprayer mixes the bleach with the water to a 10% ( 1 part bleach to 9 parts water ) solution (minimum 5,000 ppm available chlorine), so the water cartridge will need to be filled 9 times before the bleach cartridge is emptied. 

**Always remove gross dirt and debris from surface prior to disinfection.

  1. Spray Activate™ 6-8" from surface until surface is thoroughly wet.
  2. Let stand for 2 minutes.  
  3. Wipe with a clean cloth or paper towel and allow to air dry.  
  4. For food contact surfaces, rinse with potable water.  

 

Instructions for Cleaning Prior to Disinfecting against Clostridium difficile spores:

  1. Wear appropriate barrier protection such as gloves, gowns, masks or eye covering.
  2. Thoroughly clean fecal matter/waste from surfaces/objects before disinfection by application with a clean cloth, mop, and/or sponge saturated with Activate™. 
  3. Vigorously wipe and/or scrub surfaces/object until all visible soil is removed.  Pay special attention to high-touch surfaces.  Clean surfaces in patient rooms in an appropriate manner, such as from right to left or left to right on horizontal surfaces, and top to bottom on vertical surfaces, to minimize spreading of the spores.  Clean restrooms last.  Do not reuse soiled cloths.
  4. Immediately dispose of materials used in the cleaning process that may contain feces/waste in accordance with local regulations for infectious materials disposal.
  5. Disinfect: Spray Activate™ 6-8" from surface until surface is thoroughly wet; let stand 4 minutes.  Wipe with a clean, damp cloth or paper towel, or allow to air dry.

 

To Pre-Clean Instruments Prior to Terminal Sterilization:

  1. Place instruments in a suitable container.
  2. Spray Activate™ on instruments until all surfaces are thoroughly wet.
  3. Allow to stand for us to 10 minutes.
  4. Rinse instruments with water, and follow with an appropriate sterilization/high-level disinfecting process. 

NOTE:  Use Active™ on stainless steel instruments only.

 

To Disinfect Non-Critical, Pre-Cleaned Instruments:

  1. Instruments first must be thoroughly cleaned to remove gross dirt and debris, rinsed, and dried.
  2. Place instruments in a suitable container.
  3. Spray Activate™ on instruments until all surfaces are thoroughly wet.
  4. Let stand for 2 minutes.  To kill HIV-1 and HBV, allow 30 seconds contact time at room temperature. To kill Clostridium difficile spores, let stand for 4 minutes.
  5. Wipe with a clean, damp cloth or paper towel, and allow to air dry.

 

This product is not to be used as a terminal sterilant/high-level disinfectant on any surface or instrument that (1) is introduced directly into the human body, either into or in contact with the bloodstream or normally sterile areas of the body, or (2) contacts intact mucous membranes but which does not ordinarily penetrate the blood barrier or otherwise enter normally sterile areas of the body.  This product may be used to pre-clean or decontaminate critical or semi-critical medical devices prior to sterilization or high-level disinfection.

Kills HIV and HBV on pre-cleaned environmental surfaces and objects previously soiled with blood/body fluids in healthcare setting or other settings in which there is an expected likelihood of soiling of inanimate surfaces/objects with blood/body fluids, and in which the surfaces/objects likely to be soiled with blood/body fluids can be associated witht the potential for transmission of Human Immunodeficiency Virus Type-1 (HIV-1) (associated with AIDS) and Hepatitis B Virus (HBV).

 

Special Instructions for Cleaning and Decontaminating Against HIV-1 and HBV on Surfaces/Objects Soiled with Blood/Body Fluids:

  1. When handling items soiled with blood/body fluids, personal protective equipment such as disposabel gloves, gowns, masks, and eye coverings must be used.
  2. Blood/body fluids must be thoroughly cleaned from surfaces/objects prior to disinfecting.  Spray Activate™ on surface/object until all surfaces are thoroughly wet; let stand for 30 seconds.  Wipe with a clean cloth or paper towel, and allow to air dry.
  3. Blood/body fluids and cleaning materials must be autoclaved and disposed of according to federal, state, and local regulations for infectious waste disposal.

 

To Clean and Disinfect in a Veterinary Application:

  1. Remove all animals and feed from premises, animal transporation vehicles, crates, etc.
  2. Remove all litter, droppings, and manure from walls, floors, and surfaces of facilities occupied or traversed by animals.
  3. Empty all feeding and watering equipment.
  4. Pre-clean all surfaces with soap or deteregent and rinse with water.
  5. Spray Activate™ on surface/object until all surfaces are saturated: let stand for 2 minutes.  To kill HIV and HBV, allow 30 seconds at room temperature.
  6. Rinse all surfaces with potable water.
  7. Ventilate buildings and other closed spaces.  Do not house animals or employ equipment until treated surfaces have been thoroughly rinsed with water and allowed to dry.  Thoroughly scrub all treating, feeding, and watering applications with soap or detergent, and rinse with potable water before reuse.

 

Storage and Disposal of the Activate™ Bleach:

Do not contaminate food or feed by storage, disposal or cleaning of equipment.

Pesticide Storage:  This product degrades with age; test active level of sprayed product periodically with high-level chlorine test strips.  Store in a cool (50°-72°F), dry area away from direct sunlight and heat to avoid deterioration.  In case of a spill, flood area with large quantities of water.

Pesticide Disposal:  Products or rinsates that cannot be used should be diluted with water before disposal in a sanitary sewer.

Container Handling:  Non-refillable container.  Do not re-use or refill this container.  Offer for recycling if available or place in trash collection.

 

 

VioNexus™ No-Rinse Spray is an alcohol-based hand sanitizer for protection against bacteria when hands are not visibly soiled or contaminated.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining kidney and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  GATA3 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply GATA3 (Guanine-Adenine-Thymine-Adenine binding protein 3) Control Slides are for the positive immunohistochemical staining of GATA3, a transcription factor that plays a role in directing cell proliferation and development,  with expression primarily in breast and urothelial carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply GATA3 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

GATA3 positive expression  Brown nuclear staining 
Adipose Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque GATA3 (L50-823) is the concentrated primary antibody used.  Dilute primary antibody to 1/200 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque GATA3 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining kidney and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  WT1 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.
Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Wilms’ Tumor (WT1) Control Slides are for the positive immunohistochemical staining of WT1, which plays an important role in cell growth and differentiation and is expressed in kidney, malignant mesothelioma, ovarian carcinoma, gonadoblastoma, nephroblastoma (Wilms’ tumor) and acute myeloid leukemia.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply WT1 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

WT1 positive expression Brown nuclear staining
Lung Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque WT1 (6F-H2) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque WT1 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Vimentin quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Vimentin Control Slides are for the positive immunohistochemical staining of Vimentin, a ubiquitous intermediate filament found primarily in a wide variety of mesenchymal cells.  Vimentin is also commonly used to assess the effects of over-fixation and immunoreaction.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Vimentin primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Vimentin positive expression Brown cytoplasmic staining
Adipose Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Vimentin (V9) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Vimentin Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat organ and negative staining human myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Toxoplasma sp. quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Toxoplasma sp., Animal Control Slides are for the positive immunohistochemical staining of Toxoplasma, a crescent shaped sporozoan found as an intracellular parasite in various tissues.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Toxoplasma gondii primary antibody.  Incubate at room temperature for 30- 60 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Toxoplasma positive expression Golden brown/brown organisms
Myometrium Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Bio SB Toxoplasma gondii (polyclonal BDB6047) is the concentrated primary antibody used.  Dilute primary antibody to 1/200 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Bio SB Toxoplasma gondii Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining thyroid and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  TTF-1 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Thyroid Transcription Factor (TTF-1) Control Slides are for the positive immunohistochemical staining of TTF-1, selectively expressed in lung and thyroid, and aids in the classification of lung and thyroid tumors.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply TTF-1 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Link. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply Label. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

TTF-1 positive expression Brown nuclear staining
Myometrium Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Agilent TTF-1 (8G7G3/1) is the concentrated primary antibody used.  Dilute primary antibody to 1/50 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Agilent LSAB2 (K0675) Visualization Kit provides the Link and Label solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Agilent TTF-1 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Agilent LSAB2 Visualization Kit datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  CD3, T-Cell quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply CD3, T-Cell Control Slides are for the positive immunohistochemical staining of T-Cells. CD3 is considered to be a pan-T-cell marker, and widely used in detection of T-cell malignancies, both immature and mature.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply CD3 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

T-Cell positive expression Brown membrane staining
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque CD3 (MRQ-39) is the concentrated primary antibody used.  Dilute primary antibody to 1/250 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque CD3 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining pancreas and negative staining kidney.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Synaptophysin quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Synaptophysin Control Slides are for the positive immunohistochemical staining of Synaptophysin, expressed in normal neuroendocrine cells and neoplasms as well as in brain neurons, spinal cord, retina, Paneth cells in the gastrointestinal tract, and gastric parietal cells. It is considered a broad-range marker of neural and neuroendocrine differentiation.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Synaptophysin primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Synaptophysin positive expression Brown cytoplasmic staining
Kidney Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Synaptophysin (polyclonal 336A) is the concentrated primary antibody used.  Dilute primary antibody to 1/300 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Synaptophysin Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Spirochete quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Spirochete Treponema sp., Artificial Control Slides are for the positive immunohistochemical staining of spirochetes, the causative agent of a variety of diseases such as; syphilis, bejel, pinta, yaws and lyme. Treponema hyodysenteriae purchased from American Type Culture Collection is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Treponema pallidum primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply HRP-Polymer solution. Incubate for 20 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  14. Rinse slides in four changes of distilled water.
  15. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  16. Rinse slides in warm tap water to blue sections.
  17. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Spirochete positive expression Brown
Negative lung Negative for spirochetes
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare Treponema pallidum (polyclonal ACA135) is the concentrated primary antibody used.  Dilute primary antibody to 1/300 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Biocare Mach 2™ Universal HRP-Polymer Detection (M2U522) is the HRP-Polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare Treponema pallidum Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Biocare Mach 2™ Universal HRP-Polymer Detection datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining organ and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  SOX-10 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply SOX-10 (SRY-Box 10) Control Slides are for the positive immunohistochemical staining of the SOX-10 protein, expressed in melanocytes, melanomas, breast carcinomas, gliomas and schwannomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply SOX-10 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

SOX-10 positive expression Brown nuclear staining
Myometrium Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30% Aqueous Solution (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque SOX-10 (polyclonal 383A) is the concentrated primary antibody used.  Dilute primary antibody to 1/50 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque SOX-10 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining melanoma and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  S-100 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply S-100 Control Slides are for the positive immunohistochemical staining of S-100, a ubiquitous protein expressed in a number of cells. Its demonstration is of great value in the identification of several neoplasms, particularly melanomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply S-100 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

S-100 positive expression Brown cytoplasmic & nuclear staining
Lung Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque S-100 (4C4.9) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque S-100 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining prostate and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  PSA quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Prostate Specific Antigen (PSA) Control Slides are for the positive immunohistochemical staining of PSA, present in prostate tissue and expressed in the vast majority of prostate carcinomas.  It recognizes primary and metastatic prostatic neoplasms and rarely in tumors of non-prostatic origin.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply PSA primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

PSA positive expression Brown cytoplasmic staining
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque PSA (ER-PR8) is the concentrated primary antibody used.  Dilute primary antibody to 1/50 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque PSA Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  CK5/14 + p63 + P504S positive staining prostate and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  CK5/14 + p63 + P504S quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for six months from date of receipt. Revalidate after six months to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply CK5/14 + p63 + P504S Control Slides provide a useful combination of markers helpful in diagnosing prostatic intraepithelial neoplasia (PIN). For ease of screening, CK5/14 + p63 + P504S positive staining is in a single piece of prostate tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply CK5/14 + p63 + P504S multiplex primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply MACH 2 Double Stain 2 Polymer.  Incubate for 30 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  14. Rinse slides in two changes of buffer.
  15. Prepare required quantity of Vulcan Fast Red Chromogen.
  16. Tap off excess buffer; apply Vulcan Fast Red. Incubate for 10 minutes.
  17. Rinse slides in four changes of distilled water.
  18. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  19. Rinse slides in warm tap water to blue sections.
  20. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

P504S positive expression Red granular cytoplasmic staining
CK5/14 positive expression Brown cytoplasmic staining
p63 positive expression Brown nuclear staining
Myometrium Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare CK5/14 + p63 + P504S (PPM 225 DS) is the pre-diluted multiplex primary antibody used.
  5. Biocare MACH 2 Double Stain 2 Polymer Detection Kit (MRCT525) provides the polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. Biocare Vulcan Fast Red Chromogen Kit 2 (FR805) is the second chromogen used.
  8. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare CK5/14 + p63 + P504S datasheet.
  2. Biocare MACH 2 Double Stain 2 Polymer Detection Kit datasheet.
  3. Cell Marque DAB Substrate Kit datasheet.
  4. Biocare Vulcan Fast Red Chromogen Kit 2 datasheet
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  P504S and CK5/14 positive staining prostate, p63 positive staining prostate and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain: CK5/14 + p63 + P504S quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for six months from date of receipt. Revalidate after six months to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply CK5/14 + p63 + P504S, Multi-Tissue Control Slides provide a useful combination of markers helpful in diagnosing prostatic intraepithelial neoplasia (PIN).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply CK5/14 + p63 + P504S multiplex primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply MACH 2 Double Stain 2 Polymer.  Incubate for 30 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  14. Rinse slides in two changes of buffer.
  15. Prepare required quantity of Vulcan Fast Red Chromogen.
  16. Tap off excess buffer; apply Vulcan Fast Red. Incubate for 10 minutes.
  17. Rinse slides in four changes of distilled water.
  18. Counterstain lightly with Hematoxylin Stain, Gill l (Part 1180) for 5 minutes.
  19. Rinse slides in warm tap water to blue sections.
  20. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

P504S positive expression Red granular cytoplasmic staining
CK5/14 positive expression Brown cytoplasmic staining
p63 positive expression Brown nuclear staining
Myometrium Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare CK5/14 + p63 + P504S (PPM 225DS) is the pre-diluted multiplex primary antibody used.
  5. Biocare MACH 2 Double Stain 2 Polymer Detection Kit (MRCT525) provides the polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. Biocare Vulcan Fast Red Chromogen Kit 2 (FR805) is the second chromogen used.
  8. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare CK5/14 + p63 + P504S datasheet.
  2. Biocare MACH 2 Double Stain 2 Polymer Detection Kit datasheet.
  3. Cell Marque DAB Substrate Kit datasheet.
  4. Biocare Vulcan Fast Red Chromogen Kit 2 datasheet
  5. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining benign prostate and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  p63 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply p63 Control Slides are for the positive immunohistochemical staining of p63, present in normal prostate basal cells and negative in malignant prostate gland tumors. The p63 protein is expressed in the majority of squamous cell carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply p63 primary antibody.  Incubate at room temperature for 20 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply HRP-Polymer solution. Incubate for 20 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  14. Rinse slides in four changes of distilled water.
  15. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  16. Rinse slides in warm tap water to blue sections.
  17. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

p63 positive expression Brown nuclear staining
Myometrium Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare p63 (4A4) is the concentrated primary antibody used.  Dilute primary antibody to 1/50 working dilution with Biocare Van Gogh Yellow Diluent (PD902).
  5. Biocare Mach 2™ Universal HRP-Polymer Detection (M2U522) is the HRP-Polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare p63 Antibody datasheet.
  2. Biocare Van Gogh Yellow Diluent datasheet.
  3. Biocare Mach 2™ Universal HRP-Polymer Detection datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining colon and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  p53 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply p53 Control Slides are for the positive immunohistochemical staining of p53, shown to be a prognostic indicator in colorectal, breast, lung and urothelial carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply p53 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

p53 positive expression Brown nuclear staining
Adipose Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque p53 (DO7) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque p53 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining lung and negative staining kidney.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  p40 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply p40 Control Slides are for the positive immunohistochemical staining of p40, highly specific for squamous and basal cells and selectively expressed in lung squamous cell carcinoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply p40 primary antibody.  Incubate at room temperature for 20 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply HRP-Polymer solution. Incubate for 20 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  14. Rinse slides in four changes of distilled water.
  15. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  16. Rinse slides in warm tap water to blue sections.
  17. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

p40 positive expression Brown nuclear staining
Kidney Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare p40 (BC28) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Biocare Van Gogh Yellow Diluent (PD902).
  5. Biocare Mach 2™ Universal HRP-Polymer Detection (M2U522) is the HRP-Polymer solution used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare p40 Antibody datasheet.
  2. Biocare Van Gogh Yellow Diluent datasheet.
  3. Biocare Mach 2™ Universal HRP-Polymer Detection datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining breast and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  p16 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply p16 Control Slides are for the positive immunohistochemical staining of p16, a tumor suppressor protein that plays an important role in cell cycle regulation and acts as a tumor suppressor implicated in the prevention of cancers. p16 can serve as a surrogate marker for high risk HPV in cases of cervical, head and neck and a variety of HPV related carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply p16 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

p16 positive expression Brown nuclear & cytoplasmic staining
Adipose Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. BD Pharmingen p16 (G175-405) is the concentrated primary antibody used.  Dilute primary antibody to 1/20 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. BD Pharmingen p16 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining pancreas and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  NSE quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Neuron-Specific Enolase (NSE) Control Slides are for the positive immunohistochemical staining of NSE, expressed in both normal and neoplastic cells of neuronal and neuroendocrine origin, and can be used for identification of peripheral nerves, neural and neuroendocrine tumors.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Neuron-Specific Enolase primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Link.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply Label.  Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

NSE positive expression Brown cytoplasmic staining
Adipose Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Agilent Neuron-Specific Enolase (BBS/NC/VI-H14) is the primary antibody used.  Dilute primary antibody to a 1/3 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Agilent LSAB2 (K0675) Visualization Kit provides the Link and Label solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Agilent Neuron-Specific Enolase (NSE) Antibody datasheet
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Agilent LSAB2 Visualization Kit datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining normal colon.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  MLH1, MSH2, MSH6, PMS2 quality control stained slides included.

Reactivity: Guaranteed product specific reactivity for six months from date of receipt. Revalidate after six months to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply MLH1, MSH2, MSH6 and PMS2 primary antibodies.  Apply each antibody to an individual slide and tissue section.  Incubate each at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply HRP-Polymer solution. Incubate for 20 minutes.
  11. Rinse slides in two changes of buffer.
  12. Prepare required quantity of DAB substrate/chromogen.
  13. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  14. Rinse slides in two changes of buffer.
  15. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  16. Rinse slides in warm tap water to blue sections.
  17. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

MLH1 positive expression Brown nuclear staining
MSH2 positive expression Brown nuclear staining
MSH6 positive expression Brown nuclear staining
PMS2 positive expression Brown nuclear staining

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Biocare MLH1 (G168-15) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Biocare Van Gogh Yellow Diluent (PD902).
  5. Biocare MSH2 (FE11) is the concentrated primary antibody used.  Dilute primary antibody to 1/400 working dilution with Biocare Renoir Red Diluent (PD904).
  6. Biocare MSH6 (BC/44) is the concentrated primary antibody used. Dilute primary antibody to 1/250 working dilution with Biocare Van Gogh Yellow Diluent.
  7. Biocare PMS2 (A16-4) is the concentrated primary antibody used. Dilute primary antibody to 1/200 working dilution with Biocare Renoir Red Diluent.
  8. Biocare Mach 2™ Universal HRP-Polymer Detection (M2U522) is the HRP-Polymer solution used.
  9. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  10. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Biocare MLH1, MSH2, MSH6 and PMS2 Antibody datasheets.
  2. Biocare Van Gogh Yellow Diluent datasheet.
  3. Biocare Renoir Red Diluent datasheet.
  4. Biocare Mach 2™ Universal HRP-Polymer Detection datasheet.
  5. Cell Marque DAB Substrate Kit datasheet.
  6. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining melanoma and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Melan-A (MART-1) quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Melan-A (MART-1) Control Slides are for the positive immunohistochemical staining of Melan-A, expressed in melanocytes of normal skin, retina, nevi, and in the vast majority of melanomas, and is useful in establishing the diagnosis of metastatic melanomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply MART-1 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer.  Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Melan-A positive expression Brown cytoplasmic staining
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque MART-1 (A103) is the concentrated primary antibody used.  Dilute primary antibody to a 1/50 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque MART-1 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  CD45 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply CD45, Leucocyte Common Antigen (LCA) Control Slides are for the positive immunohistochemical staining of CD45, routinely used in the differential diagnosis of undifferentiated neoplasms, whenever malignant lymphoma is suspected. A positive result is highly indicative of lymphoid or myeloid origin.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply CD45 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

RESULTS:

CD45, LCA positive expression Brown membrane staining
Adipose Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque CD45 (2B11 & PD7/26) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque CD45 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Lambda quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Lambda Control Slides are for the positive immunohistochemical staining of surface and hidden determinants of lambda light chain on normal and neoplastic B-cells and expressed in B-cell follicles of human lymphoid tissue.  Lambda is an important marker to determine monoclonality of B lymphocyte neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide.  Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Lambda primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Link. Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply Label. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB.  Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Lambda positive expression Brown cytoplasmic staining
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Agilent Lambda Light Chains (polyclonal A0193) is the concentrated primary antibody used. Dilute primary antibody to a 1/1000 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Agilent LSAB2 (K0675) Visualization Kit provides the Link and Label solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Agilent Lambda Light Chains Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Agilent LSAB2 Visualization Kit datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil and negative staining adipose.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Ki-67 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Ki-67 Control Slides are for the positive immunohistochemical staining of Ki-67, a marker of proliferating cell populations and useful as a prognostic indicator in a number of neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Ki-67 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Ki-67 positive expression Brown nuclear staining
Adipose Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Ki-67 (SP6) is the concentrated primary antibody used.  Dilute primary antibody to 1/200 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Ki-67 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Kappa quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Kappa Control Slides are for the positive immunohistochemical staining of surface kappa light chain on normal and neoplastic B-cells. Kappa is an important marker to determine monoclonality of B lymphocyte neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Kappa primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Kappa positive expression Brown cytoplasmic staining
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Kappa (L1C1) is the primary antibody used.  Dilute primary antibody to a 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Kappa Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining melanoma and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  HMB-45 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply HMB-45 (Human Melanoma Black-45) Control Slides are for the positive immunohistochemical staining of HMB-45, expressed in fetal and neonatal melanocytes, junctional and blue nevus cells, and malignant melanoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply HMB-45 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

HMB-45 positive expression Brown cytoplasmic staining
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque HMB-45 (HMB-45) is the concentrated primary antibody used.  Dilute primary antibody to 1/150 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque HMB-45 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver and negative staining kidney.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Hepatitis B Surface Antigen quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Hepatitis B Surface Antigen (HBsAG) Control Slides are for the positive cytoplasmic immunohistochemical staining of Hepatitis B Surface Antigen, a protein present on the surface of the hepatitis B virus (HBV).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Hepatitis B Surface Antigen primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

HBsAG positive expression Brown cytoplasmic staining
Kidney Negative
Nuclei Blue

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Hepatitis B Surface Antigen (S1-210) is the concentrated primary antibody used.  Dilute primary antibody to 1/800 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Hepatitis B Surface Antigen Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver and negative staining kidney.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Hepatitis B Core Antigen quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Hepatitis B Core Antigen (HBcAG) Control Slides are for the positive nuclear immunohistochemical staining of Hepatitis B Core Antigen, a protein present on the core of the hepatitis B virus (HBV).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Hepatitis B Core Antigen primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill l (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

HBcAG positive expression Brown nuclear staining
Kidney Negative

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Hepatitis B Core Antigen (polyclonal 216A) is the concentrated primary antibody used.  Dilute primary antibody to 1/200 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Hepatitis B Core Antigen Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining glioblastoma and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Glial Fibrillary Acidic Protein quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Glial Fibrillary Acidic Protein (GFAP) Control Slides are for the positive immunohistochemical staining of GFAP, expressed in astrocytes, Schwann cells, satellite cells, and enteric glial cells. This marker is widely used to differentiate neoplasms of astrocytic origin from other central nervous system neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Glial Fibrillary Acidic Protein primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

GFAP positive expression Brown cytoplasmic staining
Myometrium Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Glial Fibrillary Acidic Protein (EP672Y) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Glial Fibrillary Acidic Protein Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining ductal breast and negative staining myometrium.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  E-cadherin quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply E-cadherin Control Slides are for the positive immunohistochemical staining of E-cadherin, expressed in epithelial lineage and glandular epithelium cells as well as adenocarcinoma of the lung, gastrointestinal tract, and ovary. It will also differentiate positive staining ductal vs. negative staining lobular breast carcinoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply E-cadherin primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Mayer Modified (Part 1202) for 2-5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

E-cadherin positive expression Brown membrane staining
Myometrium Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque E-cadherin (EP700Y) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque E-cadherin Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining gastrointestinal tract and negative staining lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Helicobacter sp. quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Helicobacter sp. Control Slides are for the positive immunohistochemical staining of Helicobacter sp., spiral shaped bacterium strongly associated with inflammation of the stomach and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Helicobacter Pylori primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Helicobacter sp. positive expression Golden to dark brown bacteria
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Helicobacter Pylori (polyclonal 215A) is the concentrated primary antibody used.  Dilute primary antibody to 1/750 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Helicobacter Pylori Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal gastrointestinal tract and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Helicobacter sp. quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Helicobacter sp., Animal Control Slides are for the positive immunohistochemical staining of Helicobacter sp., spiral shaped bacterium strongly associated with inflammation of the stomach and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Helicobacter Pylori primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Helicobacter sp. positive expression Golden to dark brown bacteria
Lung Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Helicobacter Pylori (polyclonal 215A) is the concentrated primary antibody used.  Dilute primary antibody to 1/750 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Helicobacter Pylori Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung and negative staining human lung.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Helicobacter sp. quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Helicobacter sp., Artificial Control Slides are for the positive immunohistochemical staining of Helicobacter sp., spiral shaped bacterium strongly associated with inflammation of the stomach and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.

Helicobacter pylori purchased from American Type Culture Collection is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Helicobacter Pylori primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Helicobacter sp. positive expression Golden to dark brown bacteria
Lung Negative
Background staining possible
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Helicobacter Pylori (polyclonal 215A) is the concentrated primary antibody used.  Dilute primary antibody to 1/750 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Helicobacter Pylori Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining gastrointestinal stromal tumor (GIST) and negative staining kidney.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  DOG1 quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply DOG1 (discovered on GIST-1) Control Slides are for the positive immunohistochemical staining of DOG1, expressed predominantly on the plasma membrane of gastrointestinal stromal tumors (GIST).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply DOG1 primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

DOG1 positive expression Brown membrane & cytoplasmic staining
Kidney Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque DOG1 (SP31) is the concentrated primary antibody used.  Dilute primary antibody to 1/25 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque DOG1 Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining leiomyoma uterus and negative staining tonsil.

Fixation: Formalin 10%, Phosphate Buffered (Part 1090).

Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.

Quality Control Stain:  Desmin quality control stained slide(s) included.

Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity.

Storage: 15-30°C in a light deprived and humidity controlled environment.

Intended Use: To verify histological techniques and reagent reactivity.

Before using unstained control slides, review the enclosed stained slide(s) to ensure that this tissue source is acceptable for testing needs.

 

APPLICATION:

Newcomer Supply Desmin Control Slides are for the positive immunohistochemical staining of Desmin, a protein found in normal smooth, skeletal, and cardiac muscle cells and expressed in leiomyoma, leiomyosarcoma, rhabdomyoma, and rhabdomyosarcoma tumors.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

  1. Heat dry sections in oven according to your laboratory protocol.
  2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each.  Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    1. See Procedure Note #1.
  3. Proceed, if necessary, with an epitope/antigen retrieval technique approved for use in your laboratory.
  4. Rinse in distilled water; tap off excess water.
  5. Circle sections with Pap Pen Liquid Blocker (Part 6505, 6506 or 6507) to reduce reagent usage and ensure tissue coverage.
  6. Block endogenous peroxidase with freshly made 3% Hydrogen Peroxide. Incubate for 5 minutes.
    1. See Procedure Note #2.
  7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
    1. See Procedure Note #3.
  8. Tap off excess buffer; apply Desmin primary antibody.  Incubate at room temperature for 30 minutes.
  9. Rinse slides in two changes of buffer.
  10. Tap off excess buffer; apply Amplifier.  Incubate for 10 minutes.
  11. Rinse slides in two changes of buffer.
  12. Tap off excess buffer; apply HRP Polymer. Incubate for 10 minutes.
  13. Rinse slides in two changes of buffer.
  14. Prepare required quantity of DAB substrate/chromogen.
  15. Tap off excess buffer; apply DAB. Incubate for 5 minutes.
  16. Rinse slides in four changes of distilled water.
  17. Counterstain lightly with Hematoxylin Stain, Gill I (Part 1180) for 5 minutes.
  18. Rinse slides in warm tap water to blue sections.
  19. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Desmin positive expression Brown cytoplasmic staining
Tonsil Negative
Nuclei Blue

 

PROCEDURE NOTES:

  1. Do not allow sections to dry out at any point during procedure.
  2. Dilute sufficient Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
  3. Dilute sufficient Tris Buffered Saline 0.05M, pH 7.6, 10X (Part 140304) with distilled water to a 1/10 solution prior to use for all buffer rinses in this procedure.
  4. Cell Marque Desmin (D33) is the concentrated primary antibody used.  Dilute primary antibody to 1/100 working dilution with Cell Marque Emerald: Antibody Diluent (936B).
  5. Cell Marque HiDef Detection™ HRP Polymer System (954D) provides the Amplifier and HRP Polymer solutions used.
  6. Cell Marque DAB Substrate Kit (957D) is the chromogen used.
  7. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

  1. Cell Marque Desmin Antibody datasheet.
  2. Cell Marque Emerald: Antibody Diluent datasheet.
  3. Cell Marque HiDef Detection™ Polymer System datasheet.
  4. Cell Marque DAB Substrate Kit datasheet.
  5. Modifications developed by Newcomer Supply Laboratory.