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 picric acid-acetone 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.

 

STAINING PROCEDURE:

1. 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. 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 blue stops running, 1-2 dips.
      1. 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 slides but not to dryness.
       1. 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); 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. 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, follow manufacturer’s recommendation for deparaffinization and clearing steps.

       

REFERENCES:

1. 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.

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

CAVIWIPES SURFACE DISINFECTION TOWELETTES FEATURES AND BENEFITS:

• Proven effective against SARS-CoV-2 on hard non-porous surfaces
• Kills TB 3 minutes and MRSA, HIV-1 and HCV in 2 minutes
• Cleaner and disinfectant in one
• Stays fully saturated – designed to help optimize fluid capacity during use
• Convenient, ready-to-use towel and CaviCide in one
• Nonabrasive – for use on hard non-porous surfaces
• Superior surface contact – durable, non-woven towels won’t bunch up during use

 

QUALIFICATIONS:

• • EPA List N Compliant

 

CAVIWIPES SURFACE DISINFECTION TOWELETTES LIST OF USES:

• Ambulance equipment surfaces
• Animal care facilities
• Bathrooms
• Correctional facilities
• Daycare Centers
• Dental Offices
• Emergency medical settings
• Emergency vehicles
• Exterior surfaces of anesthesia machines and respiratory therapy equipment
• Health club facilities
• Hospitals
• Infant/child care equipment surfaces
• Interior and exterior surfaces of infant incubators, bassinets
• Isolation areas
• Laboratories
• Laundry rooms
• Neonatal units
• Nursing homes
• Operating rooms
• Ophthalmic and optometric facilities
• Outpatient surgical centers
• Oxygen hoods
• Schools
• Surgical centers

 

CAVIWIPES SURFACE DISINFECTION TOWELETTES KILL CLAIMS:

3 Minute Efficacy Against

Mycobacterium

• Mycobacterium tuberculosis var:bovis (BCG)

Bacteria

• Pseudomonas aeruginosa
• Salmonella enterica
• Staphylococcus aureus

Fungi

• Trichophyton mentagrophytes

2 Minute Efficacy Against

Multidrug-Resistant Bacteria

• Methicillin Resistant Staphylococcus aureus (MRSA)
• Staphylococcus aureus with reduced susceptibility to vancomycin
• Vancomycin Resistant Enterococcus faecalis (VRE)

Enveloped Viruses

• Hepatitis B Virus (HBV)
• Hepatitis C Virus (HCV)
• Human Immunodeficiency Virus (HIV-1)
• Herpes Simplex Virus (type 1)
• Herpes Simplex Virus (type 2)
• Influenza A2 Virus
• SARS-CoV-2 (COVID-19 Virus)
• Human Coronavirus

 

STEINER-CHAPMAN MODIFIED SILVER STAIN 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 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
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)
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 provided staining procedure.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

1. 1. If necessary, heat dry tissue sections/slides in oven.
   2. All glassware/plasticware must be acid cleaned prior to use.
      1. 1. See Procedure Notes #1 and #2.
   3. Preheat Solution A: Zinc Formalin Sensitizer to 60°C.
   4. Preheat Solution B: Silver Nitrate 1%, Aqueous to 60°C.
   5. Prepare Hydroquinone Working Solution; combine and mix well.
      1. 1. Ingredient D: Hydroquinone, Powder 0.5 gm (or one rounded scoop with reusable mini sampling spoon)
         2. Distilled water     25 ml 

1. 6. Prepare fresh Reducing Solution by combining:
      1. 1. Solution C: Gum Mastic 2.5 %, Alcoholic 15 ml
         2. Hydroquinone Working Solution (Step #5) 25 ml
         3. Filter, then add and mix well:
         4. Solution B: Silver Nitrate 1%, Aqueous            0.6 ml
         5. Preheat solution in 45°C water bath. Save for Step #16.
   7. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

1. 8. 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. 1. See Procedure Note #3.
   9. Wash well in two changes of Acidulated Water pH 4.0-4.1 (Part 10013).
      1. 1. See Procedure Note #4.
   10. Sensitize slides in preheated Solution A: Zinc Formalin Sensitizer (Step #3) in a 60°C water bath or oven for 10 minutes.

        Microwave Modification:  See Procedure Note #5.

1. 1. 1. 1. Place slides in a plastic Coplin jar with Solution A: Zinc Formalin Sensitizer. Microwave for 1 minute at 60°C.
         2. Remove from microwave; Move to fume hood, cover and incubate for 1 minute with gentle agitation.

1. 11. Rinse in two changes of Acidulated Water pH 4.0-4.1 (Part 10013).
   12. Place slides in preheated Solution B: Silver Nitrate 1%, Aqueous (Step #4) and incubate in 60°C water bath or oven for 15 minutes.

Microwave Modification: 

1. 1. 1. 1. Place sides in plastic Coplin jar with Solution B: Silver Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
         2. Remove from microwave and let stand an additional 90 seconds; agitate occasionally for even heat distribution.

1. 13. Rinse well in several changes of distilled water.
       1. 1. Excessive rinsing may cause nuclei to pick up silver.
   14. Dip briefly in two changes of fresh 95% and 100% ethyl alcohols.
   15. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
   16. Place slides in preheated Reducing Solution (Step #6) in 45°C water bath for 15-30 minutes with frequent agitation. Examine microscopically after 15 minutes of incubation.
       1. 1. Check microscopically by dipping slide in 100% alcohol.
          2. Review for desired staining results.
          3. If necessary, return to warm solution; check every 2 minutes until results are achieved.

        Microwave Modification: 

1. 1. 1. 1. 1. Place slides in a plastic Coplin jar with Reducing Solution. Microwave for 1 minute at 70° Remove from microwave.
            2. Pipette solution twice with plastic pipette to evenly distribute heated solution.
            3. Cover and let sit for 2-4 minutes.
            4. Check microscopically by dipping slide in 100% alcohol.
            5. Review for desired staining results.
            6. If necessary, return to warm solution, check every 2 minutes until desired results are achieved.
   17. Dehydrate in two changes of 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. 1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
   2. No metals of any kind should come in contact with silver solutions to prevent precipitation of silver salts.  Use plastic forceps (Part 5500) or paraffin coated metal forceps.
   3. Drain slides after each step to prevent solution carry over.
   4. Acidulated Water pH 4.0-4.1 rinses (Steps #9 and #11) are recommended for proper tissue pH and enhanced staining.
   5. The microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   6. If using a xylene substitute follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 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	250 ml	500 ml	1 Liter	4 Liters
Hydrochloric Acid 20%, Aqueous			Part 12087A	Part 12087B
Schiff Reagent, McManus	Part 1371A	Part 1371E	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 used for the demonstration of DNA (deoxyribonucleic acid) in tissue sections.

 

METHOD:

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

1. 1. 1. See Procedure Note #1.

Technique:  Paraffin sections cut at 4 microns
Solutions:  All solutions 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. 1. If necessary, heat dry tissue sections/slides in oven.
   2. Prepare Hydrochloric Acid Working Solution; combine and mix well.
      1. 1. Hydrochloric Acid, 20% Aqueous          16 ml
         2. Distilled Water          24 ml
         3. Preheat Hydrochloric Acid Working Solution; 45 minutes at 60°C.

 

STAINING PROCEDURE:

1. 3. 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. 1. See Procedure Notes #2 and #3.
   4. Hydrolyze sections in preheated Hydrochloric Acid Working Solution (Step #2) at 60°C for 10 minutes.
      1. 1. See Procedure Notes #4 and #5.
   5. Place slides directly in Schiff Reagent, McManus for 45 minutes.
   6. Wash in running tap water for 5 minutes; rinse in distilled water.
   7. Counterstain in Light Green SF Yellowish Stain 0.2%, Aqueous (Part 12202) for 1 minute.
   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:

DNA	Red-purple
Nuclei	Red-purple
Background	Green

 

PROCEDURE NOTES:

1. 1. Bouin fixed tissue is unsatisfactory for use with Feulgen reaction.
   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. Maintain Hydrochloric Acid Working Solution at 60°C during the hydrolysis process.
   5. Prolonged exposure to hydrochloric acid may over-hydrolyze sections and destroy the reaction.
   6. If using a xylene substitute, follow manufacturer’s recommendation for the clearing steps.

 

REFERENCES:

1. 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.

OIL RED O 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 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     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 Kit procedure is classified as a physical staining method.  Oil Red O (ORO) staining is used for identification of fat in frozen tissue sections.

 

METHOD:

Fixation: Fresh tissue or formalin fixed unprocessed tissue
Technique: Frozen sections cut at 8-10 microns on adhesive slides

1. 1. See Procedure Notes #1 and #2.

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 provided staining procedure.  Some solutions in the kit may contain extra volumes.

 

PRESTAINING PREPARATION:

1. 1. Preheat Solution B: Oil Red O Stain, Propylene Glycol in 60°C oven for a minimum of 30 minutes. Save for Step #7.

(Skip if using room temperature staining method Step #6.)

1. 2. If a film of precipitate forms on Solution B: Oil Red O Stain, Propylene Glycol, skim the surface with filter paper before use.

 

STAINING PROCEDURE:

1. 3. Fix frozen sections in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
   4. Rinse sections in two changes of distilled water.
   5. Blot water and place in Solution A: Propylene Glycol 100%, ACS for 2-5 minutes with agitation.
   6. Room Temperature ORO Staining: Place in Solution B: Oil Red O Stain, Propylene Glycol for 1 hour with agitation.
   7. Heated ORO Staining: Place in preheated Solution B: Oil Red O Stain, Propylene Glycol for 7-10 minutes. Agitate occasionally.
   8. Differentiate in Solution C: Propylene Glycol 85%, Aqueous for 3 minutes with agitation.
   9. Rinse in two changes of distilled water.
   10. Counterstain in Solution D: Hematoxylin Stain, Mayer Modified for 2-3 minutes.
   11. Wash in several changes of tap water.
   12. Optional: Blue slides in Lithium Carbonate, Saturated Aqueous (Part 12215) or Scott Tap Water Substitute (Part 1380) for 10 dips.
   13. Wash in several changes of tap water.
   14. Blot water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) mounting medium.
       1. 1. See Procedure Notes #3 and #4.

 

RESULTS:

Fat:	Bright red
Nuclei:	Blue to dark blue

 

PROCEDURE NOTES:

1. 1. Freeze tissue according to laboratory protocol.
   2. To freeze formalin fixed tissue that has not been processed:
      1. 1. Wash fixed tissue in running water for 5 minutes.
         2. Blot excess water from tissue and freeze.
   3. Use minimal pressure with coverslip application to avoid displacement of fat/lipid staining.
   4. To remove trapped air bubbles or to recoverslip;
      1. 1. Soak slide in warm water until coverslip slides off.
         2. Blot water from slide.
         3. Remount with new coverslip and Mount-Quick Aqueous.

 

REFERENCES:

1. 1. Prophet, Edna B., Bob Mills, Jacquelyn Arrington and Leslie Sobin. Laboratory Methods in Histotechnology. Washington, D.C.: American Registry of Pat 1992.178.
   2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 205.
   3. Modifications developed by Newcomer Supply Laboratory.

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 counterstain that combines acid fuchsin and saturated picric acid.  The acid fuchsin selectively stains collagen and osteoid tissue red. The picric acid component stains muscle, elastin, fibrin and cytoplasm yellow.

This Van Gieson 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, Mϋller-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. 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. 1. See Procedure Notes #1 and #2.
   2. Proceed with selected stain procedure:
      1. 1. Verhoeff-Van Gieson (VVG) Elastic Stain
         2. Bile Stain, Hall’s Method
         3. Colloidal Iron, Mϋller-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. 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. 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. Picric acid element may act as a decolorizer in some procedures.
   4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 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	Part 10422
Iodine, Lugol’s, 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 product labels.

 

APPLICATION:

Newcomer Supply Twort’s Gram Stain Set provides stain solutions for the Gram Stain, Hucker-Twort, a rapid procedure for staining gram-positive and gram-negative bacteria without picric acid. The Twort Stain combines Neutral Red and Fast Green, for clear detection of red gram-negative bacteria with a green 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 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. 1. If necessary, heat dry tissue sections/slides in oven.
   2. Filter Crystal Violet-Oxalate Stain, Alcoholic (Part 10422) with high quality filter paper.

 

STAINING PROCEDURE:

1. 3. 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. 1. See Procedure Note #1.
   4. Stain in freshly filtered Crystal Violet-Oxalate Stain, Alcoholic, (Step #2) for 30 seconds.
   5. Rinse quickly in distilled water.
   6. Mordant in Iodine, Lugol’s, Aqueous (Part 12092) for 20 seconds.
   7. Rinse quickly in distilled water.
   8. Decolorize individually with Acetone, ACS (Part 10014); 2 quick dips.
      1. 1. Or until tissue remains light gray.
   9. Rinse quickly in distilled water.
   10. Prepare fresh Twort Stain; combine and mix well.
       1. 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.
   11. Stain in fresh Twort Stain for 2 minutes.
   12. Rinse quickly in distilled water; carefully blot dry.
   13. Agitate slides quickly in clean Acetone, ACS to remove excess stain and dehydrate (do not use any alcohols).
       1. 1. The use of alcohols will remove Neutral Red.
   14. 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. 1. Drain slides after each step to prevent solution carry over.
   2. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 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.

Tech Memo 2: Toluidine Blue Stain for Mohs Technique

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:

• • As part of a staining system for blood smears (Click here for Differential Stain Kit Part #9112B)

 

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

 

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 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. 1. 1. See Procedure Note #1.

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

 

STAINING PROCEDURE: METHOD FOR NEUTRAL FATS

1. 1. Examine a fresh fecal sample collected, stored and prepared according to laboratory protocol.
      1. 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. 1. Prepare in advance 36% Acetic Acid, Glacial; mix well.
      1. 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 a fresh fecal sample collected, stored and prepared according to laboratory protocol.
      1. 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 hold slide over preheated hot plate until bubbles appear; quickly remove slide; reheat two additional times.
      1. 1. See Procedure Note #3.
   8. Immediately coverslip and examine microscopically while warm.

 

RESULTS:

Fatty acids

Red/orange refractile globules

 

PROCEDURE NOTES:

1. 1. For optimum results fresh, unpreserved fecal material is required.
   2. Refer to laboratory protocols 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. 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.

SOLUTION:

	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	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 since 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. 1. 1. See Procedure Notes #1and #2.

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. 1. Fix frozen section slides in Formalin 10%, Phosphate Buffered (Part 1090) for 1 minute.
   2. Rinse sections 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. 1. See Procedure Note #3.
   5. Differentiate in Propylene Glycol 85%, Aqueous (Part 133912) for 3 minutes with agitation.
   6. Rinse in distilled water.
   7. Counterstain in Nuclear Fast Red Stain, Kernechtrot (Part 1255) for 5 minutes.
      1. 1. Shake solution well before use; do not filter.
   8. Wash gently in several changes of tap water.
      1. 1. See Procedure Note #4.
   9. Blot excess water from slide; coverslip with Mount-Quick Aqueous (Part 6271A) mounting medium.
      1. 1. See Procedure Notes #5 and #6.

 

RESULTS:

Fat	Blue-black
Nuclei	Red

 

PROCEDURE NOTES:

1. 1. Freeze tissue according to laboratory protocol.
   2. To freeze formalin fixed tissue that has not been processed:
      1. 1. Wash tissue in running tap water for 5 minutes.
         2. Blot excess water from tissue and freeze.
   3. To decrease staining time; preheat Sudan Black B Stain, Propylene Glycol in a 60°C oven; stain for 3-10 minutes.
   4. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in sections.
   5. Use minimal pressure with coverslip application to avoid displacement of fat/lipid staining.
   6. To remove trapped air bubbles or to recoverslip;
      1. 1. Soak slide in warm water until coverslip falls off.
         2. Blot excess water from slide.
         3. Remount with new coverslip and Mount-Quick Aqueous.

 

REFERENCES:

1. 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 is 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. 1. Prepare a well-made smear 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. 1. See Procedure Note #1.
   4. Place slides on flat staining rack suspended over sink.
      1. 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, ACS (Part 10014) until color stops running; 5-10 seconds.
      1. 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. 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. 1. Gentle heat or methanol are accepted methods of smear fixation.
   2. Do not allow smears to dry out at any point during 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.
   4. For enhanced counterstaining;
      1. 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.
         4. Rinse well in distilled water.

       

REFERENCES:

1. 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.

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 product labels.

 

APPLICATION:

Newcomer Supply Grocott Methenamine Silver Set, GMS provides the silver solutions for the Fungus GMS stain procedure.  This is the best staining method 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 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. 1. If necessary, heat dry tissue sections/slides in oven.
   2. All glassware/plasticware must be acid cleaned prior to use.
      1. 1. See Procedure Notes #1 and #2.
   3. Prepare Silver-Methenamine Working Solution and mix well:
      1. 1. Solution A: Silver Nitrate                 20 ml
         2. Solution B: Methenamine Borate 20 ml
   4. Preheat Silver-Methenamine Working Solution to 45°C-60°C in a water bath approximately 20-30 minutes before use.
      1. 1. Maintain solution between 45°C-60°C to minimize precipitate.
         2. Do not preheat if using Microwave Modification; Step 11.

 

STAINING PROCEDURE:

1. 5. 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. 1. See Procedure Notes #3 and #4.
   6. Oxidize in Chromic Acid 5%, Aqueous (Part 10341) for 1 hour.

        Microwave Modification: See Procedure Note #5.

1. 1. 1. 1. Microwave Solution A: Chromic Acid 5%, Aqueous without slides in a plastic Coplin jar (Part 5184) for 1 minute at 60°C.  Add slides to heated Solution A and oxidize for 10 minutes.

1. 7. Wash well in running tap water; rinse in distilled water.
   8. Place in Sodium Bisulfite 1%, Aqueous (Part 13821) for 1 minute.
   9. Wash for 5 minutes in running tap water; rinse well in distilled water.
   10. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
       1. 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.

1. 11. Microwave Modification:
       1. 1. Incubate slides in a plastic Coplin jar with Silver-Methenamine Working Solution. Microwave for 5 minutes at 45°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.
   12. Rinse in three to four changes of distilled water.
       1. 1. Do not use tap water at this step.
   13. Tone in Gold Chloride 0.1%, Aqueous (Part 11285) until sections turn gray; 10-30 seconds.
   14. Rinse well in distilled water.
   15. Remove unreduced silver in Sodium Thiosulfate 2%, Aqueous (Part 13888) for 2 minutes.
   16. Wash in running tap water for 5 minutes; rinse in distilled water.
   17. Counterstain in Light Green SF Yellowish Stain 0.02%, Aqueous (Part 12204) for 2 minutes.
   18. 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

 

PROCEDURE NOTES:

1. 1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
   2. No metals of any kind should come in contact with silver solutions to prevent precipitation of silver salts. Use plastic forceps (Part 5500) or paraffin coated metal forceps.
   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. The microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 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:

SOLUTION: 

	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 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.  Van Gieson Stain serves 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. 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. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Place slides in freshly filtered Fouchet Reagent for 5 minutes.
   5. Wash in three changes of tap water; rinse in distilled water.
   6. Stain sections in Van Gieson Stain (Part 1404) for 5 minutes.
   7. Rinse quickly in 95% ethyl alcohol.
   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:

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

 

PROCEDURE NOTES:

1. 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. 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.

 

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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Factor XIIIa (EP3372) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Factor XIIIa Antibody datasheet.
   2. 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 expressed in a number of malignant lymphomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque EMA (E29) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque EMA Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque BCL2 (124) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque BCL2 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

 

PRODUCT SPECIFICATIONS:

Tissue:  Gram positive staining rat lung and gram negative staining rat lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Brown-Brenn 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-Brenn, Stain Kit:		Part 9123A	Individual Stain Solution
Solution A:	Crystal Violet-Oxalate Stain, Alcoholic	250 ml	Part 10422
Solution B:	Iodine, Gram, Aqueous	250 ml	Part 1140
Solution C:	Acetone-Alcohol 1:1	250 ml	Part 10016
Solution D:	Basic Fuchsin Stain 0.25%, Aqueous	250 ml	Part 1011
Solution E:	Tartrazine Stain 0.25%, Acetic Aqueous	250 ml	Part 14016

 

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 are used to produce the positive controls.

 

PRESTAINING PREPARATION:     

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Filter Solution A: Crystal Violet-Oxalate Stain, Alcoholic.

 

STAINING PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Stain in freshly filtered Solution A: Crystal Violet-Oxalate Stain, Alcoholic (Step #2) for 1 minute.
   5. Rinse well in distilled water.
   6. Mordant in Solution B: Iodine, Gram, Aqueous for 1 minute.
   7. Rinse well in distilled water, removing excess iodine.
   8. Decolorize in Solution C: Acetone-Alcohol 1:1 until blue stops running; 7-10 dips.
   9. Rinse well in distilled water.
   10. Place in Solution D: Basic Fuchsin Stain 0.25%, Aqueous for 90 seconds.
   11. Rinse well in distilled water.
   12. Dip once in Solution C: Acetone-Alcohol 1:1.
   13. Counterstain in Solution E: Tartrazine Stain 0.25%, Acetic Aqueous for 5-15 seconds.
   14. Rinse well in distilled water.
   15. Dehydrate in two changes of 100% ethyl alcohol, 5 dips each. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
       1. 1. Do not use 95% alcohol in the dehydration step.

 

RESULTS:

Gram negative bacteria	Red
Gram positive bacteria	Blue/violet
Background tissue	Yellow

 

PROCEDURE NOTES:

1. 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, follow manufacturer’s recommendation for deparaffinization and clearing steps.

       

REFERENCES:

1. 1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 312-313.
   2. Brown, J.H. and L. Brenn. “A Method for the Differential Staining of Gram Positive and Gram Negative Bacteria in Tissue Sections.”Bulletin of The Johns Hopkins 48.2 (1931): 69-73.
   3. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 188-189.
   4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Brown-Brenn 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-Brenn Stain Kit:		Part 9123A	Individual Stain Solution
Solution A:	Crystal Violet-Oxalate Stain, Alcoholic	250 ml	Part 10422
Solution B:	Iodine, Gram, Aqueous	250 ml	Part 1140
Solution C:	Acetone-Alcohol 1:1	250 ml	Part 10016
Solution D:	Basic Fuchsin Stain 0.25%, Aqueous	250 ml	Part 1011
Solution E:	Tartrazine Stain 0.25%. Acetic Aqueous	250 ml	Part 14016

 

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 a single tissue section.

Escherichia coli and Staphylococcus aureus purchased from Remel Microbiology Products are used to produce the positive controls.

 

PRESTAINING PREPARATION:    

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Filter Solution A: Crystal Violet-Oxalate Stain, Alcoholic.

 

STAINING PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Stain in freshly filtered Solution A: Crystal Violet-Oxalate Stain, Alcoholic (Step #2) for 1 minute.
   5. Rinse well in distilled water.
   6. Mordant in Solution B: Iodine, Gram, Aqueous for 1 minute.
   7. Rinse well in distilled water, removing excess iodine.
   8. Decolorize in Solution C: Acetone-Alcohol 1:1 until blue stops running; 7-10 dips.
   9. Rinse well in distilled water.
   10. Place in Solution D: Basic Fuchsin Stain 0.25%, Aqueous for 90 seconds.
   11. Rinse well in distilled water.
   12. Dip once in Solution C: Acetone-Alcohol 1:1.
   13. Counterstain in Solution E: Tartrazine Stain 0.25%, Acetic Aqueous for 5-15 seconds.
   14. Rinse well in distilled water.
   15. Dehydrate in two changes of 100% ethyl alcohol, 5 dips each. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
       1. 1. Do not use 95% alcohol in the dehydration step.

 

RESULTS:

Gram positive bacteria	Blue/violet
Gram negative bacteria	Red
Background tissue	Yellow

 

PROCEDURE NOTES:

1. 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, follow manufacturer’s recommendation for deparaffinization and clearing steps.

       

REFERENCES:

1. 1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 312-313.
   2. Brown, J.H. and L. Brenn. “A Method for the Differential Staining of Gram Positive and Gram Negative Bacteria in Tissue Sections.”Bulletin of The Johns Hopkins 48.2 (1931): 69-73.
   3. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 188-189.
   4. Modifications developed by Newcomer Supply Laboratory.

 

FEATURES OF THE BLOCK STORAGE SYSTEM, PLASTIC 6 DRAWER UNIT:

• • Made of ABS
  • Six segmented, impact resistant drawers
  • Seven segmented rows per drawer.  Each row includes a sponge block to support blocks in upright position.
  • Identification writing area on each drawer
  • Stackable cabinets have locking features to each other
  • Each 6 drawer unit accommodates up to 2,100 FFPE tissue block cassettes
  • Cabinet Dimension: 17 3/8″L x 9 1/2″W x 15 1/4″H
  • Interfaces with other popular plastic block storage systems!

 

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. 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. 1. See Procedure Note #1.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #2 and #3.
   4. Prepare fresh Ferrocyanide Working Solution directly before use; combine and mix well.
      1. 1. Solution A: Hydrochloric Acid 20%, Aqueous        20 ml
         2. Solution B: Potassium Ferrocyanide 10%, Aqueous  20 ml
   5. Place slides in fresh Ferrocyanide Working Solution for 20 minutes.
   6. Rinse in three changes of tap water; rinse in distilled water.
   7. Place in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
      1. 1. Shake solution well before use; do not filter.
   8. Rinse well in distilled water.
      1. 1. See Procedure Note #4.
   9. 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. 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, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 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.

 

PRODUCTS:

	Each	Case of 12
Activate™ Bleach Sprayer with Refillable 11 Ounce Water Cartridge	Part 5101
Activate™ Bleach Cartridge Refills, 11 Ounce, Disposable		Part 5102

 

Additionally Needed:

Activate™ High-Level Chlorine Test Strips

Part 5100

 

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

 

APPLICATION:

Newcomer Supply Activate™ Bleach Sprayer with Refillable 11 Ounce Water Cartridge is a non-aerosol sprayer system that maintains bleach and water in separate containers and automatically mixes and dispenses a 10% bleach solution every time the sprayer trigger is pulled.

• • • Ensures the bleach solution is continually fresh and effective.
    • Prevents bleach from degrading, always a 10% solution.
    • Eliminates the hazard of mixing bleach solutions by hand.
    • Fast contact time for practical disinfecting.
    • Each Activate™ Bleach Cartridge Refill, 11 Ounce, Disposable, makes approximately one gallon of 10% bleach.
    • Safe and effective for healthcare, research and veterinary facilities.

 

ACTIVATE™ BLEACH SPRAYER PROCEDURE:

1. 1. Open locking tab on water side of the sprayer; remove water bottle.
   2. Fill water bottle with cool tap water and insert sprayer tube.
   3. Re-install water bottle onto sprayer; firmly close locking tab.
   4. Remove threaded plastic cap from bleach sprayer cartridge.
      1. 1. Do not puncture or tamper with plug in the neck of the bleach sprayer cartridge.
   5. Open locking tab on bleach side of the sprayer.
   6. Install bleach cartridge; firmly close the locking tab.
   7. Point bleach sprayer into the sink, pump trigger 20 times to prime.
   8. Once primed the bleach sprayer is ready to use.
   9. Hold sprayer 6 to 8 inches from surface to be cleaned/disinfected.
      1. 1. Pre-clean surfaces of loose dirt and debris.
         2. See Procedure Notes #1 and #2.
   10. Spray Activate™ bleach until all surfaces are thoroughly wet.
   11. Allow to react for 2 to 4 minutes or minimum contact times:
       1. 1. Viricide: Allow a minimum of 30 seconds contact time at room temperature (HIV-1 and HBV)
          2. Bactericide: Allow a minimum of 2 minutes contact time at room temperature (MRSA and Bacteria)
          3. Sporicide: Allow a minimum of 4 minutes contact time at room temperature (Clostridium difficile)
   12. Wipe surface with a clean cloth or paper towels; air dry.
       1. 1. Rinse food and animal contact surfaces with potable water.
   13. Refill water bottle or replace bleach cartridge as needed.
       1. 1. Sprayer will stop if bleach or water bottles are empty.
          2. Water bottle will require 9 refills before a single bleach cartridge is emptied.
          3. Prime sprayer after installation of a new bleach cartridge.
   14. Periodically test sprayed bleach solution for available chlorine with Activate™ High-Level Chlorine Test Strips (5100).

 

PROCEDURE NOTES:

1. 1. Activate™ bleach solution is safe on hard, non-porous surfaces such as porcelain, tile, marble, plastic, glass and colorfast surfaces.
   2. Use Activate™ bleach solution on stainless steel instruments only.
      1. 1. Prolonged contact with metal surfaces may cause discoloration or pitting.
         2. Rinse metal surfaces with water after application.
   3. Do not use or mix with other cleaning chemicals.
   4. For additional product information and instructions refer to Newcomer Supply website.

 

REFERENCES:

1. 1. Dapson, Janet Crookham and Richard Dapson. Hazardous Materials in the Histopathology Laboratory: Regulations, Risks, Handling, and Disposal. 4th ed. Battle Creek, MI: Anatech, 2005. 231.
   2. Rutala, William A. and David J. Weber. “Guideline for Disinfection and Sterilization in Healthcare Facilities, 2008.” (2008). CDC Public Health Publications.
   3. Modifications developed by Newcomer Supply Laboratory.

 

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).

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. To kill HIV-1 and HBV, allow 30 seconds contact time at room temperature.
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. Personal Protection: Wear appropriate barrier protection such as gloves, gowns, masks or eye covering.
2. Cleaning Procedure: Thoroughly clean fecal matter/waste from surfaces/objects before disinfection by application with a clean cloth, mop, and/or sponge saturated with Activate™. 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.
3. Infectious Materials Disposal: Immediately dispose of materials used in the cleaning process that may contain feces/waste in accordance with local regulations for infectious materials disposal.
4. Disinfection Procedure: 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 STERLILIZATION:

1. Place instruments in a suitable container.
2. Spray Activate™ on instruments until all surfaces are thoroughly wet.
3. Allow to stand for up to 10 minutes.
4. Rinse instruments with water, and follow with an appropriate sterilization/high-level disinfecting process.
5. Cleaning of critical and semi-critical devices must be followed by an appropriate terminal sterilization/high-level disinfection 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 with the potential for transmission of Human Immunodeficiency Virus Type-1 (HIV-1) (associated with AIDS) and Hepatitis B Virus (HBV).

 

SPECIAL INSTRUCTIONS FOR CLEANING & 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 disposable 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, or apply with a clean cloth, mop or sponge saturated with Activate™; 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. Use to clean and disinfect hard, non-porous surfaces such as feeding and watering equipment, cages, utensils, instruments, kennels, stables, catteries, etc.
2. Remove all animals and feed from premises, animal transportation vehicles, crates, etc.
3. Remove all litter, droppings, and manure from walls, floors, and surfaces of facilities occupied or traversed by animals.
4. Empty all feeding and watering equipment.
5. Pre-clean all surfaces with soap or detergent and rinse with water.
6. Spray Activate™ on surface/object until all surfaces are saturated, or apply with a clean cloth, mop or sponge saturated with Activate™, let stand for 2 minutes.  To kill HIV-1 and HBV, allow 30 seconds at room temperature.
7. Rinse all surfaces with potable water.
8. 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 diluted 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 Handwash is an alcohol-based hand sanitizer for protection against bacteria when hands are not visibly soiled. VioNexus encourages proper hand hygiene practice by making it convenient, efficient, and comfortable.

 

VIONEXUS NO-RINSE SPRAY HANDWASH FEATURES AND BENEFITS:

• Meets CDC Guidelines for Hand Hygiene in Healthcare Settings
• Kills 99.9% of bacteria on skin without running water
• Contains 72% ethanol, killing germs in seconds
• Emollients help prevent dry, irritated skin and leave hands feeling soft
• No water or towels needed, eliminating cross contamination
• Fast drying for quick donning of gloves

 

VIONEXUS NO-RINSE SPRAY HANDWASH LIST OF USES:

• Animal care facilities
• Break rooms
• Community health centers
• Correctional facilities
• Dental offices
• Dialysis clinics
• Dining areas
• Donning rooms
• Emergency medical settings
• Employee work stations
• Entrances and exits
• Extended care
• General practices
• High-traffic areas
• Hospitals
• Isolation areas
• Laboratories
• Laundry rooms
• Long-term care
• Meeting rooms
• Military bases
• Neonatal units
• Nursing homes
• Operating rooms
• Ophthalmic and optometric facilities
• Orthodontist offices
• Outpatient surgical centers
• Reception desks
• Schools
• Surgical centers
• Transaction counters
• Waiting rooms

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.  Expression is primarily found in breast and urothelial carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque GATA3 (L50-823) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque GATA3 Antibody datasheet.
   2. 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, a gene involved in cell growth and differentiation. WT1 is expressed in kidney, malignant mesothelioma, ovarian serous carcinoma and nephroblastoma (Wilms’ tumor).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque WT1 (6F-H2) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque WT1 Antibody datasheet.
   2. 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 variety of mesenchymal cells.  Vimentin is also used to assess the effects of over-fixation and immunoreaction.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Vimentin (V9) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Vimentin Antibody datasheet.
   2. 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 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, 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Toxoplasma gondii 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Bio SB Toxoplasma gondii Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Bio SB Toxoplasma gondii Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque TTF-1 (EP229) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, closely follow the manufacturer’s recommendations for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Agilent TTF-1 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD3 (MRQ-39) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD3 Antibody datasheet.
   2. 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. Synaptophysin is considered a broad-range marker of neural and neuroendocrine differentiation.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Synaptophysin (MRQ-40) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Synaptophysin Antibody datasheet.
   2. 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, 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

Spirochete positive expression	Brown
Negative lung	Negative for spirochetes
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare Treponema pallidum Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare Treponema pallidum Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30% Aqueous Solution (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque SOX-10 (EP268) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque SOX-10 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque S-100 (4C4.9) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque S-100 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque PSA (ER-PR8) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque PSA Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CK5/14 + p63 + P504S 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 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. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   19. Rinse slides in warm tap water to blue sections.
   20. Air-dry slides. Dip dried slides in xylene; 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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare CK5/14 + p63 + P504S Cocktail is the primary antibody used along with Biocare detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare CK5/14 + p63 + P504S datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CK5/14 + p63 + P504S 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 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. 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 with Hematoxylin Stain, Gill l (Part 1180); 1-10 dips.
   19. Rinse slides in warm tap water to blue sections.
   20. Air-dry slides. Dip dried slides in xylene; 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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare CK5/14 + p63 + P504S Cocktail is the primary antibody used along with Biocare detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare CK5/14 + p63 + P504S datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

p63 positive expression	Brown nuclear staining
Myometrium	Negative

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare p63 (4A4) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare p63 Antibody datasheet.
   2. 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, a prognostic indicator in colorectal, breast, lung and urothelial carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque p53 (DO7) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque p53 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

p40 positive expression	Brown nuclear staining
Kidney	Negative

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare p40 (BC28) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare p40 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. BD Pharmingen p16 (G175-405) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. BD Pharmingen p16 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Agilent Neuron-Specific Enolase (BBS/NC/VI-H14) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Agilent Neuron-Specific Enolase (NSE) Antibody datasheet
   2. 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.

 

APPLICATION:

Newcomer Supply Mismatch Repair (MMR) Positive Control Slides provide a single tissue source that expresses positive reactivity in each of the MMR panel of four markers; MLH1, MSH2, MSH6 and PMS2.  Mismatch Repair testing is useful in screening for colorectal carcinoma (CRC), Microsatellite Instability (MSI) and Lynch Syndrome (LS).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 two changes of buffer.
   17. Counterstain with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare MLH1 (G168-15) is the primary antibody used.
   5. Biocare MSH2 (FE11) is the primary antibody used.
   6. Biocare MSH6 (44) is the primary antibody used.
   7. Biocare PMS2 (A16-4) is the primary antibody used.
   8. Cell Marque detection and ancillary reagents are used.
   9. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare MLH1, MSH2, MSH6 and PMS2 Antibody datasheets.
   2. 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 majority of melanomas, and is useful in establishing the diagnosis of metastatic melanomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque MART-1 (M2-7C10) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque MART-1 Antibody datasheet.
   2. 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.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD45 (2B11 & PD7/26) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD45 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Agilent Lambda Light Chains Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Agilent Lambda Light Chains Antibody datasheet.
   2. 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 a useful prognostic indicator in a number of neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Ki-67 (SP6) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Ki-67 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Kappa (L1C1) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Kappa Antibody datasheet.
   2. 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 immature melanosomes, junctional nevus cells, and melanomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque HMB-45 (HMB-45) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque HMB-45 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Hepatitis B Surface Antigen (A10F1) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Hepatitis B Surface Antigen Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining brain 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 astrocytoma, glioblastoma, ependymoma and oligodendroglioma. This marker is widely used to differentiate neoplasms of astrocytic origin from other central nervous system neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Glial Fibrillary Acidic Protein (EP672Y) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Glial Fibrillary Acidic Protein Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver 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.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque E-cadherin (EP700Y) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque E-cadherin Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining gastrointestinal tract biopsy 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 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 Control Slides are for the positive immunohistochemical staining of spiral shaped Helicobacter  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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Helicobacter pylori Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Helicobacter pylori Antibody datasheet.
   2. 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 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, Animal Control Slides are for the positive immunohistochemical staining of spiral shaped Helicobacter 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Helicobacter pylori Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Helicobacter pylori Antibody datasheet.
   2. 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 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, Artificial Control Slides are for the positive immunohistochemical staining of spiral shaped Helicobacter 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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 positive expression	Golden to dark brown bacteria
Lung	Negative
	Background staining possible
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Helicobacter pylori Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Helicobacter pylori Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque DOG1 (SP31) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque DOG1 Antibody datasheet.
   2. 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. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Desmin (D33) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Desmin Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining lung 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:  Cytomegalovirus 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 Cytomegalovirus (CMV) Control Slides are for the positive immunohistochemical staining of cytomegalovirus, a member of the herpes virus group, including herpes simplex virus types 1 and 2, varicella zoster virus, and Epstein Barr virus.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Cytomegalovirus 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CMV positive expression	Brown nuclear staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CMV (8B1.2, 1G5.2, 2D4.2) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CMV Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining skin 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:  Cytokeratin, HMW 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 Cytokeratin, HMW (High Molecular Weight) Control Slides are for the positive immunohistochemical staining of Cytokeratin, HMW, expressed in squamous, ductal and complex epithelia. Useful in differentiation of benign prostate glands from prostatic adenocarcinoma and classification of neoplastic tissue as carcinoma or epithelial origin.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Cytokeratin, HMW 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

Cytokeratin, HMW positive expression	Brown cytoplasmic staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Cytokeratin, HMW (AE3) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Cytokeratin, HMW Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining skin, positive staining colon and negative staining cardiac muscle.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Cytokeratin AE1/AE3 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 Cytokeratin AE1/AE3 Control Slides are for the positive immunohistochemical staining of Cytokeratin’s AE1/AE3, present in normal and abnormal tissues, used to distinguish epithelial carcinoma from non-epithelial malignancies and aid in epithelial tumor classification.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Cytokeratin Cocktail (AE1/AE3) 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

Cytokeratin AE1/AE3 positive expression	Brown cytoplasmic staining
Cardiac muscle	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Cytokeratin Cocktail (AE1/AE3) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Cytokeratin Cocktail (AE1/AE3) Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining colon 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:  Cytokeratin 20 (CK20) 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 Cytokeratin 20 (CK20) Control Slides are for the positive immunohistochemical staining of Cytokeratin 20, expressed in gastric and intestinal epithelium, urothelium and Merkel cells. CK20 has been identified in colon, stomach, pancreas and biliary system adenocarcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Cytokeratin 20 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CK20 positive expression	Brown cytoplasmic staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Cytokeratin 20 (Ks20.8) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Cytokeratin 20 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining pancreas 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:  Cytokeratin 7 (CK7) 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 Cytokeratin 7 (CK7) Control Slides are for the positive immunohistochemical staining of CK7, reacting with proteins in ductal, glandular and transitional epithelia of the urinary tract and bile duct. CK7 will also distinguish positive staining lung and breast epithelium from negative staining colon and prostate epithelial cells.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Cytokeratin 7 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CK7 positive expression	Brown cytoplasmic staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Cytokeratin 7 (OV-TL 12/30) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Cytokeratin 7 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil and negative staining liver.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Cytokeratin 5/6 (CK5/6) 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 Cytokeratin 5/6 (CK5/6) Control Slides are for the positive immunohistochemical staining of CK5/6, a useful marker in identification of mesothelioma and lung squamous cell carcinoma and also expressed in a wide range of normal tissue such as breast, prostate, mesothelium, skin and esophagus.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CK5/6 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CK5/6 positive expression	Brown cytoplasmic staining
Liver	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CK5/6 (D5/16B4) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CK5/6 Antibody datasheet.
   2. 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:  Chromogranin A 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 Chromogranin A Control Slides are for the positive immunohistochemical staining of Chromogranin A, expressed in a wide variety of endocrine tissues such as pituitary, pancreas, hypothalamus, and parathyroid and useful in identification of neuroendocrine tumors.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Chromogranin A 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

Chromogranin A positive expression	Brown cytoplasmic staining
Kidney	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Chromogranin A (LK2H10) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Chromogranin A Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining colon 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:  CDX2 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 CDX2 Control Slides are for the positive immunohistochemical staining of CDX2, a caudal-related homeobox gene expressed in nuclei of intestinal epithelial cells and in primary and metastatic colorectal carcinomas. CDX2 has also been identified in intestinal metaplasia of the stomach and intestinal-type gastric cancer.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CDX2 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CDX2 positive expression	Brown nuclear staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30% Aqueous Solution (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CDX2 (EPR2764Y) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CDX2 Antibody datasheet.
   2. 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:  CD138 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 CD138 Control Slides are for the positive immunohistochemical staining of CD138, a specific marker for identification of plasma cells and plasma cell differentiation within hematolymphoid tissues in benign and neoplastic conditions.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD138 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD138 positive expression	Brown membranous staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES: 

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare CD138 (B-A38) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare CD138 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil 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:  CD68 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 CD68 Control Slides are for the positive immunohistochemical staining of CD68, a useful marker for cells of macrophage lineage, including monocytes and histiocytes. Kupffer cells and osteoclasts are also stained.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD68 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD68 positive expression	Brown cytoplasmic & membrane staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30% Aqueous Solution (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD68 (Kp-1) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD68 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining thyroid 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:  CD56 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 CD56 Control Slides are for the positive immunohistochemical staining of CD56, expressed in a variety of normal and abnormal tissues.  CD56 recognizes two proteins of the neural cell adhesion molecule, the basic molecule expressed on most neuroectodermally derived tissues and neoplasms.  It is also expressed on some mesodermally derived tumors.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD56 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD56 positive expression	Brown membrane staining
Lung	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD56 (123C3.D5) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD56 Antibody datasheet.
   2. 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:  CD34 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 CD34 Control Slides are for the positive immunohistochemical staining of CD34, found in vascular endothelium and expressed by spindle cell tumors, such as solitary fibrous tumors and gastrointestinal stromal tumors (GIST). CD34 is useful for differentiation in myeloid and lymphoid neoplasms.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD34 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD34 positive expression	Brown membrane staining
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD34 (QBEnd/10) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD34 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining Hodgkin’s lymphoma 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:  CD30 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 CD30 Control Slides are for the positive immunohistochemical staining of CD30, expressed by Reed-Sternberg cells in classic Hodgkin lymphoma, the majority of anaplastic large cell lymphomas, primary cutaneous CD30 positive T-cell lymphoproliferative disorders, and in embryonal carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD30 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD30 positive expression	Brown cellular membrane staining
Kidney	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD30 (Ber-H2) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD30 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining GIST (gastrointestinal stromal tumor) 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:  CD117, c-Kit 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 CD117, c-Kit Control Slides are for the positive immunohistochemical staining of CD117.  CD117 is found in a wide variety of tumors and is useful in classification of tumors of the gastrointestinal tract and seminomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD117 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD117 positive expression	Brown cytoplasmic & membrane staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD117, c-kit (YR145) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD117 Antibody datasheet.
   2. 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:  CD10 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 CD10 Control Slides are for the positive immunohistochemical staining of CD10, also known as Common Acute Lymphoblastic Leukemia Antigen (CALLA).  CD10 is useful in the classification of B-cell leukemias and lymphomas and identification of hepatocellular and renal cell carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD10 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD10 positive expression	Brown cytoplasmic & membrane staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD10 (56C6) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD10 Antibody datasheet.
   2. 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:  CD8 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 CD8 Control Slides are for the positive immunohistochemical staining of CD8, a useful marker for the detection of T-cell involvement in cytotoxic immunoreactions and classification of lymphocytes and malignant lymphomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD8 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CD8 positive expression	Brown membranous staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD8 (C8/144B) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD8 Antibody datasheet.
   2. 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:  Carcinoembryonic Antigen (CEA) 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 Carcinoembryonic Antigen (CEA) Control Slides are for the positive immunohistochemical staining of CEA, useful in distinguishing adenocarcinoma from mesothelioma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CEA 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

CEA positive expression	Brown cytoplasmic staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CEA Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CEA Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining mesothelioma 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:  Calretinin 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 Calretinin Control Slides are for the positive immunohistochemical staining of Calretinin, useful in differentiating mesothelioma from adenocarcinomas of the lung and other sources.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Calretinin 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

Calretinin positive expression	Brown cytoplasmic & nuclear staining
Myometrium	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Biocare Calretinin (polyclonal) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Biocare Calretinin Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver 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:  Alpha-Fetoprotein (AFP) 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, Alpha-Fetoprotein (AFP) Control Slides are for the positive immunohistochemical staining of Alpha-Fetoprotein, a secretory protein expressed in liver carcinomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply Alpha-Fetoprotein 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

AFP positive expression	Brown cytoplasmic staining
Adipose	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque Alpha-Fetoprotein Polyclonal is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque Alpha-Fetoprotein Antibody datasheet.
   2. 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:  BCL6 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 BCL6 (B-cell lymphoma 6) Control Slides are for the positive immunohistochemical staining of BCL6, commonly expressed in diffuse large cell lymphomas, follicular lymphomas and Burkitt’s lymphoma.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply BCL6 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

BCL6 positive expression	Brown nuclear staining
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque BCL6 (GI191E/A8) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque BCL6 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining tonsil 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:  CD20 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 CD20, B-Cell Control Slides are for the positive immunohistochemical staining of normal and neoplastic B-cells, generally located in follicles of lymph nodes and tonsils. CD20 is considered to be a pan-B-cell marker, occasionally detected in T-cell malignancies and a very strong marker of mature B-cell lymphomas.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Note #1.
   3. Proceed 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. 1. See Procedure Note #2.
   7. Wash slides gently in distilled water. Rinse in two changes of Tris Buffered Saline.
      1. 1. See Procedure Note #3.
   8. Tap off excess buffer; apply CD20 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 with Hematoxylin Stain, Gill I (Part 1180); 1-10 dips.
   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:

B-cell positive expression	Brown cytoplasmic staining
Kidney	Negative
Nuclei	Blue

 

PROCEDURE NOTES:

1. 1. Do not allow sections to dry out at any point during procedure.
   2. Dilute Hydrogen Peroxide 30%, Aqueous (Part 1206) with distilled water to a 3% (1/10) solution prior to use.
   3. Dilute 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.
   4. Cell Marque CD20 (L26) is the primary antibody used along with Cell Marque detection and ancillary reagents.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Cell Marque CD20 Antibody datasheet.
   2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining uterus
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Actin, Smooth Muscle 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 Actin, Smooth Muscle, Control Slides are for the positive immunohistochemical staining of smooth muscle actin. Myofibroblasts and myoepithelial cells will also positively react.

 

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 Actin, Smooth Muscle 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:

Actin, Smooth Muscle 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 Actin, Smooth Muscle (1A4) 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 Actin, Smooth Muscle 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 gouty tophi tissue.
Fixation: Alcohol, Ethyl Denatured, 100% (Part 10841)
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Gomori Methenamine Silver 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 Urate Stain, Gomori Methenamine Silver Method:	Individual Stain Solution
Methenamine 3%, Aqueous	Part 12239
Silver Nitrate 5%, Aqueous	Part 13805
Sodium Borate 5%, Aqueous	Part 13826
Gold Chloride 0.25%, Aqueous	Part 11287
Sodium Thiosulfate 2.5%, Aqueous	Part 13889
Light Green SF Yellowish Stain 0.2%, Aqueous	Part 12202

 

APPLICATION:

Newcomer Supply Urates Control Slides are for the positive histochemical staining of urates in tissue sections. With abnormal accumulations found around joints and in soft tissues, this disturbance in uric acid metabolism is known as gout, with collections of urate crystals referred to as gouty tophi.

Calcium pyrophosphate (CPP) crystals, known as pseudogout, may be demonstrated with this procedure. When viewed with a polarizing filter and red compensator filter, gout and pseudogout can be distinguished.

Fixation: Urate crystals are soluble in aqueous solutions. Fix in Alcohol, Ethyl Denatured, 100%; a minimum of two changes, 4 hours each.

Processing:  Transfer from 100% ethyl alcohol fixative to xylene for 1 hour; proceed with equal parts xylene/paraffin at 58°C for 2 hours. Infiltrate with paraffin for a minimum of 1 hour; embed.

Technique: Chill paraffin blocks in 100% ethyl alcohol; cut paraffin sections with minimal water bath exposure.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. All glassware/plasticware must be acid cleaned prior to use.
      1. 1. See Procedure Notes #1 and #2.
   3. Prepare Methenamine Silver Stock Solution.
      1. 1. Methenamine 3%, Aqueous                 50 ml
         2. Silver Nitrate 5%, Aqueous                5 ml
         3. Slowly add silver nitrate; mix to clear milky precipitate.
         4. Store clear solution at 2°-8°C for up to 2 months.
   4. Prepare Methenamine Silver Working Solution; combine, mix well.
      1. 1. Methenamine Silver Stock Solution 25 ml
         2. Distilled Water 25 ml
         3. Sodium Borate 5%, Aqueous                   3 ml
   5. Preheat Methenamine Silver Working Solution to 60°C in a water bath.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 6. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Rinse in two changes of 100% ethyl alcohol, 10 dips each.
      1. 1. Do not use 95% ethyl alcohol or distilled water steps.
         2. See Procedure Notes #3 and #4.

1. 7. Incubate slides in preheated Methenamine Silver Working Solution (Step #5) in a 60°C water bath for 30 minutes.
      1. 1. Remove control slide, rinse in warm distilled water, check microscopically for adequate silver development. Crystals should be dark brown/black.
         2. If structures are not sufficiently dark, place slides back in warm silver solution.
         3. Recheck at 2-3 minute intervals until desired intensity is achieved.
   8. Rinse well in distilled water.
   9. Tone in Gold Chloride 0.25%, Aqueous until brown colorization disappears; 5 to 30 seconds.
   10. Rinse well in distilled water.
   11. Place in Sodium Thiosulfate 2.5%, Aqueous; 2 to 3 minutes.
   12. Wash well in running tap water for 3 minutes; rinse in distilled water.
   13. Counterstain in Light Green SF Yellowish Stain 0.2%, Aqueous for 1 to 2 minutes, depending on preference of counterstain intensity.
   14. 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:

Light Field Microscopy:
	Gout/urate crystals	Black
	Background	Green
Polarized/Red Compensator Filter: (long axes aligned parallel)
	Gout/urate crystals	Yellow, long & needle shaped
	Pseudogout crystals	Blue, shorter & rhomboidal

 

PROCEDURE NOTES:

1. 1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
   2. No metals of any kind should come in contact with silver solutions to prevent precipitation of silver salts. Use plastic forceps (Part 5500) or paraffin coated metal forceps.
   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. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.255-256, 267-268.
   2. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 187-188.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 225-226.
   4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining uterus.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Gomori One-Step Aniline 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 Trichrome, Gomori One-Step, Aniline Blue Stain Kit:		Part 9176B/A	Individual Stain Solution
Solution A:	Bouin Fluid	250/500 ml	Part 1020
Solution B:	Ferric Chloride, Acidified	125/250 ml	Part 1409
Solution C:	Hematoxylin 1%, Alcoholic	125/250 ml	Part 1409
Solution D:	Trichrome Stain, Gomori One-Step, Aniline Blue	250/500 ml	Part 1403
Solution E:	Acetic Acid 0.5%, Aqueous	250/500 ml	Part 100121

 

APPLICATION:

Newcomer Supply Trichrome, Uterus Control Slides are for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Preheat Solution A: Bouin Fluid to 56°-60°C in oven or water bath. (Skip if using overnight method or microwave procedure.)

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Mordant in preheated Solution A: Bouin Fluid (Step #2) for one hour at 56°-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
      1. 1. Skip Step #4 if tissue was originally Bouin fixed.

       Microwave Modification:  See Procedure Note #3.

1. 1. 1. 2. Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid. Microwave for 5 minutes at 60°C.

1. 5. Wash well in running tap water; rinse in distilled water.
   6. Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
      1. 1. Solution B: Ferric Chloride, Acidified 20 ml
         2. Solution C: Hematoxylin 1%, Alcoholic 20 ml

1. 7. Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
   8. Wash in running tap water for 10 minutes; rinse in distilled water.
      1. 1. See Procedure Note #4.
   9. Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
   10. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
   11. Rinse quickly 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:

Collagen and mucin	Blue
Muscle fibers, cytoplasm and keratin	Red
Nuclei	Blue/black

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
   5. Trichrome, Uterus Control Slides can be used as positive controls with any Trichrome procedure.
   6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
   4. Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
   5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining liver, positive staining kidney and positive staining uterus.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Gomori One-Step Aniline 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 Trichrome, Gomori One-Step, Aniline Blue Stain Kit:		Part 9176B/A	Individual Stain Solution
Solution A:	Bouin Fluid	250/500 ml	Part 1020
Solution B:	Ferric Chloride, Acidified	125/250 ml	Part 1409
Solution C:	Hematoxylin 1%, Alcoholic	125/250 ml	Part 1409
Solution D:	Trichrome Stain, Gomori One-Step, Aniline Blue	250/500 ml	Part 1403
Solution E:	Acetic Acid 0.5%, Aqueous	250/500 ml	Part 100121

 

APPLICATION:

Newcomer Supply Trichrome, Multi-Tissue Control Slides, use a combination of tissue sources for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Preheat Solution A: Bouin Fluid to 56°-60°C in oven or water bath. (Skip if using overnight method or microwave procedure.)

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Mordant in preheated Solution A: Bouin Fluid (Step #2) for one hour at 56°-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
      1. 1. Skip Step #4 if tissue was originally Bouin fixed.

       Microwave Modification:  See Procedure Note #3.

1. 1. 1. 2. Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid. Microwave for 5 minutes at 60°C.

1. 5. Wash well in running tap water; rinse in distilled water.
   6. Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
      1. 1. Solution B: Ferric Chloride, Acidified 20 ml
         2. Solution C: Hematoxylin 1%, Alcoholic 20 ml
   7. Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
   8. Wash in running tap water for 10 minutes; rinse in distilled water.
      1. 1. See Procedure Note #4.
   9. Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
   10. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
   11. Rinse quickly 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:

Collagen and mucin	Blue
Muscle fibers, cytoplasm and keratin	Red
Nuclei	Blue/black

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
   5. Trichrome, Multi-Tissue Control Slides can be used as positive controls with any Trichrome procedure.
   6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
   4. Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
   5. 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:  Gomori One-Step Aniline 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 Trichrome, Gomori One-Step, Aniline Blue Stain Kit:		Part 9176A/B	Individual Stain Solution
Solution A:	Bouin Fluid	250/500 ml	Part 1020
Solution B:	Ferric Chloride, Aqueous	125/250 ml	Part 1409
Solution C:	Hematoxylin 1%, Alcoholic	125/250 ml	Part 1409
Solution D:	Trichrome Stain, Gomori One-Step, Aniline Blue	250/500 ml	Part 1403
Solution E:	Acetic Acid 0.5%, Aqueous	250/500 ml	Part 100121

                         

APPLICATION:

Newcomer Supply Trichrome, Liver Control Slides are for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Preheat Solution A: Bouin Fluid to 56°-60°C in oven or water bath. (Skip if using overnight method or microwave procedure.)

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Mordant in preheated Solution A: Bouin Fluid (Step #2) for one hour at 56°-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
      1. 1. Skip Step #4 if tissue was originally Bouin fixed.

       Microwave Modification:  See Procedure Note #3.

1. 1. 1. 2. Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid. Microwave for 5 minutes at 60°C.

1. 5. Wash well in running tap water; rinse in distilled water.
   6. Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
      1. 1. Solution B: Ferric Chloride, Aqueous 20 ml
         2. Solution C: Hematoxylin 1%, Alcoholic 20 ml
   7. Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
   8. Wash in running tap water for 10 minutes; rinse in distilled water.
      1. 1. See Procedure Note #4.
   9. Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
   10. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
   11. Rinse quickly 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:

Collagen and mucin	Blue
Muscle fibers, cytoplasm and keratin	Red
Nuclei	Blue/black

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
   5. Trichrome Liver Control Slides can be used as positive controls with any Trichrome procedure.
   6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
   4. Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
   5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining kidney.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Gomori One-Step Aniline 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 Trichrome, Gomori One-Step, Aniline Blue Stain Kit:		Part 9176B/A	Individual Stain Solution
Solution A:	Bouin Fluid	250/500 ml	Part 1020
Solution B:	Ferric Chloride, Acidified	125/250 ml	Part 1409
Solution C:	Hematoxylin 1%, Alcoholic	125/250 ml	Part 1409
Solution D:	Trichrome Stain, Gomori One-Step, Aniline Blue	250/500 ml	Part 1403
Solution E:	Acetic Acid 0.5%, Aqueous	250/500 ml	Part 100121

 

APPLICATION:

Newcomer Supply Trichrome, Kidney Control Slides are for the positive histochemical staining of connective tissue and to differentially demonstrate collagen and muscle fibers.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Preheat Solution A: Bouin Fluid to 56°-60°C in oven or water bath. (Skip if using overnight method or microwave procedure.)

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Mordant in preheated Solution A: Bouin Fluid (Step #2) for one hour at 56°-60°C or overnight at room temperature. Cool at room temperature for 5-10 minutes.
      1. 1. Skip Step #4 if tissue was originally Bouin fixed.

       Microwave Modification:  See Procedure Note #3.

1. 1. 1. 2. Place slides in a plastic Coplin jar containing Solution A: Bouin Fluid. Microwave for 5 minutes at 60°C.

1. 5. Wash well in running tap water; rinse in distilled water.
   6. Prepare fresh Weigert Iron Hematoxylin; combine and mix well.
      1. 1. Solution B: Ferric Chloride, Acidified 20 ml
         2. Solution C: Hematoxylin 1%, Alcoholic 20 ml
   7. Stain in fresh Weigert Iron Hematoxylin for 10 minutes.
   8. Wash in running tap water for 10 minutes; rinse in distilled water.
      1. 1. See Procedure Note #4.
   9. Stain with Solution D: Trichrome Stain, Gomori One-Step, Aniline Blue for 20 minutes.
   10. Differentiate in Solution E: Acetic Acid 0.5%, Aqueous; 2 minutes.
   11. Rinse quickly 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:

Collagen and mucin	Blue
Muscle fibers, cytoplasm and keratin	Red
Nuclei	Blue/black

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If Weigert Iron Hematoxylin is not completely washed from tissue sections, nuclear and cytoplasmic staining may be compromised.
   5. Trichrome, Kidney Control Slides can be used as positive controls with any Trichrome procedure.
   6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Brown, Richard. Histologic Preparations: Common Problems and Their Solutions. Northfield, Ill.: College of American Pathologists, 2009. 95-101.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 165-166.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 191-192.
   4. Vacca, Linda L. Laboratory Manual of Histochemistry. New York: Raven Press, 1985. 308-310.
   5. Modifications developed by Newcomer Supply Laboratory.

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:  Steiner-Steiner Modified 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 Steiner-Steiner Modified Silver Stain Kit:		Part 9171A	Individual Stain Solution
Solution A:	Uranyl Nitrate 1%, Aqueous	250 ml	Part 14036
Solution B:	Silver Nitrate 1%, Aqueous	250 ml	Part 13804
Solution C:	Gum Mastic 2.5%, Alcoholic	350 ml	Part 1145
Ingredient D:	Hydroquinone, Powder	5 grams	Part 12089

 

APPLICATION:

Newcomer Supply Spirochete, Animal Control Slides are for the positive histochemical staining of spirochetes, the causative agent of a variety of diseases such as; syphilis, bejel, pinta, yaws and lyme.  These control slides are not validated for immunohistochemistry (IHC) use.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. All glassware/plasticware must be acid cleaned prior to use.
      1. 1. See Procedure Notes #1 and #2.
   3. Preheat Solution A: Uranyl Nitrate 1%, Aqueous to 60°C in a water bath. Save for Step #9.
   4. Preheat Solution B: Silver Nitrate 1%, Aqueous to 60°C in a water bath. Save for Step #11.
   5. Prepare Hydroquinone Solution; combine and mix well.
      1. 1. Ingredient D: Hydroquinone, Powder                0.5 gm (or one rounded scoop with reusable mini sampling spoon)
         2. Distilled Water                 25 ml

1. 6. Prepare fresh Reducing Solution by combining in order listed.
      1. 1. Hydroquinone Solution (Step #5) 25 ml
         2. Solution C: Gum Mastic 2.5%, Alcoholic         15 ml
         3. Solution B: Silver Nitrate 1%, Aqueous    0.6 ml
         4. Solution will turn milky white after addition of Gum Mastic.
         5. Preheat solution in 45°C water bath. Save for Step #15.
   7. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

1. 8. 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. 1. See Procedure Note #3.
   9. Sensitize in preheated Solution A: Uranyl Nitrate 1%, Aqueous (Step #3) for 10 minutes in a 60°C water bath.

        Microwave Modification: See Procedure Note #4.

1. 1. 1. 1. Place slides in a plastic Coplin jar with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.

1. 10. Rinse well in several changes of distilled water.
   11. Place slides in preheated Solution B: Silver Nitrate 1%, Aqueous (Step #4) and incubate in a 60°C water bath for 15 minutes.

        Microwave Modification:

1. 1. 1. 1. Place slides in a plastic Coplin jar with Solution B: Silver Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
         2. Remove from microwave, cover and let sit for 1 minute.

1. 12. Rinse well in several changes of distilled water.
       1. 1. Excessive rinsing may cause nuclei to pick up silver.

1. 13. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
   14. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
   15. Place slides in preheated Reducing Solution (Step #6) in 45°C water bath for 10-30 minutes with frequent agitation. Examine microscopically after 10 minutes of incubation.
       1. 1. Check microscopically by dipping slide in 100% alcohol.
          2. Review for desired staining results.
          3. If necessary, return to warm solution; check every 2-5 minutes until desired result is achieved.

        Microwave Modification: 

1. 1. 1. 1. Place slides in a plastic Coplin jar with Reducing Solution. Microwave for 1 minute at 70°C. Remove from microwave.
         2. Pipette solution twice with plastic pipette to evenly distribute heated solution.
         3. Cover and let sit for 1 minute.
         4. Check microscopically by dipping slide in 100% alcohol.
         5. Review for desired staining results.
         6. If necessary, return to warm solution, check every 1 minute until desired results are achieved.

1. 16. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Spirochetes	Dark brown to black
Background	Golden brown

 

PROCEDURE NOTES:

1. 1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
   2. No metals of any kind should come in contact with silver solutions to prevent precipitation of silver salts. Use plastic forceps (Part 5500) or paraffin coated metal forceps.
   3. Drain slides after each step to prevent solution carry over.
   4. The microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Garvey, Winsome. “Some Favorite Silver Stains.” The Journal of Histotechnology 19.3 (1996): 269-278.
   2. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 218-219.
   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. Swisher, Billie. “Modified Steiner Procedure for Microwave Staining of Spirochetes and Nonfilamentous Bacteria.” The Journal of Histotechnology 10.4 (1987): 241-243.
   5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining rat lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Steiner-Steiner Modified 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 Steiner-Steiner Modified Silver Stain Kit:		Part 9171A	Individual Stain Solution
Solution A:	Uranyl Nitrate 1%, Aqueous	250 ml	Part 14036
Solution B:	Silver Nitrate 1%, Aqueous	250 ml	Part 13804
Solution C:	Gum Mastic 2.5%, Alcoholic	350 ml	Part 1145
Ingredient D:	Hydroquinone, Powder	5 grams	Part 12089

 

APPLICATION:

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

Brachyspira hyodysenteriae, purchased from American Type Culture Collection, is used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. All glassware/plasticware must be acid cleaned prior to use.
      1. 1. See Procedure Notes #1 and #2.
   3. Preheat Solution A: Uranyl Nitrate 1%, Aqueous to 60°C in a water bath. Save for Step #9.
   4. Preheat Solution B: Silver Nitrate 1%, Aqueous to 60°C in a water bath. Save for Step #11.
   5. Prepare Hydroquinone Solution; combine and mix well.
      1. 1. Ingredient D: Hydroquinone, Powder                0.5 gm (or one rounded scoop with reusable mini sampling spoon)
         2. Distilled Water                 25 ml

1. 6. Prepare fresh Reducing Solution by combining in order listed.
      1. 1. Hydroquinone Solution (Step #5) 25 ml
         2. Solution C: Gum Mastic 2.5%, Alcoholic         15 ml
         3. Solution B: Silver Nitrate 1%, Aqueous  0.6 ml
         4. Solution will turn milky white after addition of Gum Mastic.
         5. Preheat solution in 45°C water bath. Save for Step #15.
   7. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

1. 8. 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. 1. See Procedure Note #3.
   9. Sensitize in preheated Solution A: Uranyl Nitrate 1%, Aqueous (Step #3) for 10 minutes in a 60°C water bath.

        Microwave Modification: See Procedure Note #4.

1. 1. 1. 1. Place slides in a plastic Coplin jar with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.

1. 10. Rinse well in several changes of distilled water.
   11. Place slides in preheated Solution B: Silver Nitrate 1%, Aqueous (Step #4) and incubate in a 60°C water bath for 15 minutes.

        Microwave Modification:

1. 1. 1. 1. Place slides in a plastic Coplin jar with Solution B: Silver Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
         2. Remove from microwave, cover and let sit for 1 minute.

1. 12. Rinse well in several changes of distilled water.
       1. 1. Excessive rinsing may cause nuclei to pick up silver.
   13. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
   14. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 5 minutes.
   15. Place slides in preheated Reducing Solution (Step #6) in 45°C water bath for 10-30 minutes with frequent agitation. Examine microscopically after 10 minutes of incubation.
       1. 1. Check microscopically by dipping slide in 100% alcohol.
          2. Review for desired staining results.
          3. If necessary, return to warm solution; check every 2-5 minutes until desired results are achieved.

        Microwave Modification: 

1. 1. 1. 1. Place slides in a plastic Coplin jar with Reducing Solution. Microwave for 1 minute at 70°C.  Remove from microwave.
         2. Pipette solution twice with plastic pipette to evenly distribute heated solution.
         3. Cover and let sit for 1 minute.
         4. Check microscopically by dipping slide in 100% alcohol.
         5. Review for desired staining results.
         6. If necessary, return to warm solution, check every 1 minute until desired results are achieved.

1. 16. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Spirochetes	Dark brown to black
Background	Golden brown

 

PROCEDURE NOTES:

1. 1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
   2. No metals of any kind should come in contact with silver solutions to prevent precipitation of silver salts. Use plastic forceps (Part 5500) or paraffin coated metal forceps.
   3. Drain slides after each step to prevent solution carry over.
   4. The microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Garvey, Winsome. “Some Favorite Silver Stains.” The Journal of Histotechnology 19.3 (1996): 269-278.
   2. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 218-219.
   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. Swisher, Billie. “Modified Steiner Procedure for Microwave Staining of Spirochetes and Nonfilamentous Bacteria.” The Journal of Histotechnology 10.4 (1987): 241-243.
   5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining striated muscle.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  PTAH 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 Phosphotungstic Acid Hematoxylin (PTAH) Stain Kit:		Part 9111A	Individual Stain Solution
Solution A:	Zenker Fixative, Modified, Zinc Chloride	250 ml	Part 1461
Solution B:	Acetic Acid, Glacial, ACS	25 ml	Part 10010
Solution C:	Potassium Permanganate 0.25%, Aqueous	250 ml	Part 133931
Solution D:	Oxalic Acid 5%, Aqueous	250 ml	Part 1293
Solution E:	Phosphotungstic Acid Hematoxylin (PTAH) Stain, Modified Mallory	250 ml	Part 1334

 

APPLICATION:

The Newcomer Supply Phosphotungstic Acid Hematoxylin (PTAH) Control Slides are for the positive histochemical demonstration of muscle striations and collagen in tissue sections.

 

PRESTAINING PREPARATION:

1. 1. Heat dry sections in oven according to your laboratory protocol.
   2. Prepare Zenker Fixative Working Solution; combine and mix well.

1. 1. 1. Solution A: Zenker Fixative, Modified, Zinc Chloride       38 ml
      2. Solution B: Acetic Acid, Glacial, ACS                                2 ml

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 3. 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. 1. See Procedure Notes #1 and #2.
   4. Fix in Zenker Fixative Working Solution (Step #2) at 56°C; 3 hours.

        Microwave Modification:  See Procedure Note #3.

1. 1. 1. 1. Place slides in a plastic Coplin jar containing Zenker Fixative Working Solution. Microwave 5 minutes at 60°

1. 5. Wash well in three changes of tap water; rinse in distilled water.
   6. Place in Solution C: Potassium Permanganate 0.25%, Aqueous for 10 minutes.
   7. Wash in three changes of tap water; rinse in distilled water.
   8. Place in Solution D: Oxalic Acid 5%, Aqueous for 10 minutes.
   9. Wash in three changes of tap water; rinse in distilled water.
   10. Stain in Solution E: PTAH Stain, Modified Mallory for 12-24 hours at room temperature, or 2 hours at 56°
       1. 1. See Procedure Note #4.

Microwave Modification:

1. 1. 1. 2. Place slides in a plastic Coplin jar containing Solution E: PTAH Stain, Modified Mallory. Microwave 7 minutes at 70°

1. 11. 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.
       1. 1. Dehydrate quickly, alcohol may extract stain from sections.

 

RESULTS:

Muscle striations, fibrin, keratin	Dark blue
Collagen, cartilage, elastic fibers	Deep reddish brown
Nuclei	Blue

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. Adjust staining time according to preference of intensity. Suggested staining time at 37°C is 18 hours.
   5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008.130-131.
   2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 178-180, 201-202.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 193-194.
   4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining colon.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Mayer Mucicarmine 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 Mucin, Mayer Mucicarmine Stain Kit:		Part 9151A/B	Individual Stain Solution
Solution A:	Ferric Chloride, Aqueous	125/250 ml	Part 1409
Solution B:	Hematoxylin 1%, Alcoholic	125/250 ml	Part 1409
Solution C:	Mucicarmine Stock Stain, Mayer	125/125 ml	Part 1250
Solution D:	Metanil Yellow Stain, Aqueous	250/500 ml	Part 12235

 

APPLICATION:

Newcomer Supply Mucin Mucicarmine Control Slides are for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin).

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

1. 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. 1. See Procedure Notes #1 and #2.
   3. Prepare fresh Weigert Iron Hematoxylin Working Solution directly before use; combine and mix well:
      1. 1. Solution A: Ferric Chloride, Aqueous   20 ml
         2. Solution B: Hematoxylin 1%, Alcoholic             20 ml
   4. Stain in fresh Weigert Iron Hematoxylin Working Solution for 7 minutes.
   5. Rinse in running tap water for 10 minutes.
   6. Prepare fresh Mayer Mucicarmine Working Solution; combine and mix well:
      1. 1. Solution C: Mucicarmine Stock Stain, Mayer   10 ml  
         2. Tap Water (do not use distilled water)   30 ml    
   7. Stain in fresh Mayer Mucicarmine Working Solution for 60 minutes or longer if a more intense stain is desired.

Microwave Modification:  See Procedure Note #3.

1. 1. 1. 1. Place slides in a plastic Coplin jar containing fresh Mayer Mucicarmine Working Solution. Microwave for 10 minutes at 70°C.

1. 8. Rinse in several changes of tap water.
   9. Counterstain in Solution D: Metanil Yellow Stain, Aqueous for 30 to 60 seconds.
   10. Dehydrate quickly through 95% and 100% ethyl alcohols. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins	Deep rose to red
Nuclei	Black
Other tissue elements	Yellow

 

PROCEDURE NOTES:

1. 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 microwave procedure was tested using a laboratory-grade microwave oven. This procedure is a guideline and techniques should be developed for use in your laboratory.
   4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

1. 1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 174-175.
   2. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 142-144.
   3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 168-169.
   4. Modifications developed by Newcomer Supply Laboratory.