PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine and positive staining lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Movat-Russell Pentachrome 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 Movat-Russell Modified Pentachrome Stain Kit: Part 9150A Individual Stain Solution
Solution A: Alcian Blue Stain 1%, Aqueous 250 ml
Solution B: Ammonium Hydroxide 28-30%, ACS 50 ml Part 1006
Solution C: Hematoxylin 10%, Alcoholic 100 ml
Solution D: Ferric Chloride 10%, Aqueous 100 ml Part 10856
Solution E: Iodine, Verhoeff, Aqueous 100 ml Part 1209
Solution F: Ferric Chloride 2%, Aqueous 250 ml Part 108553
Solution G: Sodium Thiosulfate 5%, Aqueous 250 ml Part 1389
Solution H: Crocein Scarlet 7B Stain, Aqueous 250 ml
Solution I: Acid Fuchsin Stain, Aqueous 100 ml
Solution J: Phosphotungstic Acid 5%, Aqueous 500 ml Part 13345
Solution K: Orange G Stain 1%, Aqueous 250 ml
Solution L: Acetic Acid 0.5%, Aqueous 500 ml Part 100121

 

APPLICATION:

Newcomer Supply Movat-Russell Pentachrome Control Slides are for the positive histochemical staining of connective tissue elements, mucin, fibrin, elastic fibers, muscle, and collagen.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Stain in Solution A: Alcian Blue Stain 1%, Aqueous for 20 minutes.
    4. Wash in running tap water for 5 minutes.
    5. Prepare fresh Alkaline Alcohol Solution; combine and mix well.
        1. Solution B: Ammonium Hydroxide 28-30%   5 ml
        2. Alcohol, Ethyl Denatured, 95% (Part 10842)               45 ml
    6. Place slides in fresh Alkaline Alcohol Solution for 15 minutes.
    7. Wash in running tap water for 10 minutes; rinse in distilled water.
        1. See Procedure Note #3.
    8. Prepare fresh Hematoxylin Working Stain Solution just before use in the order given; combine and mix well.
        1. Solution C: Hematoxylin 10%, Alcoholic   10 ml
        2. Alcohol, Ethyl Denatured 100% (Part 10841)   10 ml
        3. Solution D: Ferric Chloride 10%, Aqueous   10 ml
        4. Solution E: Iodine, Verhoeff, Aqueous                   10 ml
    9. Stain in fresh Hematoxylin Working Stain Solution for 15 minutes.
        1. Discard after successful differentiation in Step #11.
    10. Rinse in several changes of distilled water.
    11. Differentiate one slide at a time in Solution F: Ferric Chloride 2%, Aqueous until elastic fibers contrast sharply with the background; approximately 5-10 dips.
        1. See Procedure Note #4.
    12. Rinse in distilled water.
    13. Place in Solution G: Sodium Thiosulfate 5%, Aqueous for 1 minute.
    14. Wash in running tap water for 5 minutes; rinse in distilled water.
    15. Prepare Crocein Scarlet-Acid Fuchsin Solution:
        1. Solution H: Crocein Scarlet 7B Stain, Aqueous   40 ml
        2. Solution I: Acid Fuchsin Stain, Aqueous   10 ml
    1. Stain in Crocein Scarlet-Acid Fuchsin Solution for 1 minute.
    2. Rinse in several changes of distilled water.
    3. Rinse in Solution L: Acetic Acid 0.5%, Aqueous for 30 seconds.
    4. Place in Solution J: Phosphotungstic Acid 5%, Aqueous; two changes of 5 minutes each.
    5. Rinse in Solution L: Acetic Acid 0.5%, Aqueous.
    6. Stain in Solution K: Orange G Stain 1%, Aqueous for 15 minutes.
    7. Dehydrate through three changes of 100% ethyl alcohol, 10 dips each.   Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.
        1. Do not use 95% ethyl alcohol in the dehydration step.

 

RESULTS:

Nuclei and elastic fibers Black
Collagen and reticular fibers Yellow
Ground substance and mucin Blue
Fibrinoid, fibrin Intense red
Muscle Red

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. Completely remove Alkaline Alcohol Solution with running tap water. Failure to do so will inhibit subsequent staining steps.
    4. If background is completely colorless, the section may be over-differentiated. Over-differentiated sections can be restained in Hematoxylin Working Stain Solution (Step #9) provided sections have not been treated with an alcohol/dehydration step.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 172-174.
    2. Movat, Henry, “Demonstration of All Connective Tissue Elements in a Single ” AMA Archives of Pathology. 1955; 60 (3): 289–295.
    3. Russell H. K. Jr. “A Modification of Movat’s Pentachrome Stain.” AMA Archives of Pathology.1972; 94 (2): 187–191.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

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

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

 

CONTROL SLIDE VALIDATION:

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

 

APPLICATION:

Newcomer Supply Melanin Control Slides are for the positive histochemical staining of melanin in tissue sections.

 

PRESTAINING PREPARATION:

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

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

 

RESULTS:

Melanin Black
Nuclei Pink-red

 

PROCEDURE NOTES:

    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. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
    6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 286-287.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 276-277.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal tumor.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Toluidine 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 Toluidine Blue Stain for Mast Cells: Individual Stain Solution
Toluidine Blue Stain 0.1%, Aqueous Part 14027

 

APPLICATION:

Newcomer Supply Mast Cell, Animal Control Slides are for the positive histochemical staining of mast cells, cells filled with basophilic granules associated with inflammation and allergic reactions, which stain metachromatically with toluidine blue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:      

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

 

RESULTS:

Mast cells Deep rose-violet
Background Blue

 

PROCEDURE NOTES:

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

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.188.
    2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining bladder.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Toluidine 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 Toluidine Blue Stain for Mast Cells: Individual Stain Solution
Toluidine Blue Stain 0.1%, Aqueous Part 14027

 

APPLICATION:

Newcomer Supply Mast Cell Control Slides are for the positive histochemical staining of mast cells, cells filled with basophilic granules associated with inflammation and allergic reactions, which stain metachromatically with toluidine blue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:          

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

 

RESULTS:

Mast cells Deep rose-violet
Background Blue

 

PROCEDURE NOTES:

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

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009.188.
    2. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining spinal cord.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 8 microns on Superfrost™ Plus slides.
Quality Control Stain:  Luxol Fast 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 Luxol Fast Blue (LFB) Stain Set: Part 12218A/B Individual Stain Solution
Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic 500/1000 ml
Solution B: Lithium Carbonate, Saturated Aqueous 500/1000 ml Part 12215
Hematoxylin Stain, Harris Modified Part 1201
Acid Alcohol 1% Part 10011

   

APPLICATION:

Newcomer Supply Luxol Fast Blue (LFB) Control Slides are for the histochemical staining of myelin sheath in central nervous system tissue and peripheral nerve. PAS, cresyl violet, hematoxylin and eosin stains can be added to the LFB procedure for enhanced staining.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Prepare Working Lithium Carbonate 0.05%; combine and mix well;
        1. Solution B: Lithium Carbonate, Saturated Aqueous 5 ml
        2. Distilled Water                     95 ml

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.
        1. Stop at 95% ethyl alcohol; no distilled water rinse.
        2. See Procedure Notes #1 and #2.
    2. Incubate slides in Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic for 2 hours at 60°C or overnight at 37°C; seal lids tightly.
        1. To enhance stain, add 0.4 ml of Acetic Acid, Glacial, ACS (Part 10010) to 40 ml of Solution A before use.

        Microwave Modification: See Procedure Note #3.

        1. Place slides in a plastic Coplin jar with Solution A: Luxol Fast Blue Stain 0.1%, Alcoholic. Microwave 10 minutes at 70°C.
    1. Rinse slides quickly in 95% ethyl alcohol; 2-3 dips.
    2. Rinse slides in distilled water.
    3. Differentiate slides individually in Working Lithium Carbonate 0.05% (Step #2) for 10-15 seconds with agitation until gray and white matter are colorless and contrast with stained tissue.
    4. Further differentiate in 70% ethyl alcohol (Part 10844), until gray and white matter can be distinguished. Do not over differentiate.
    5. Rinse slides in distilled water.
    6. Check slides microscopically. Continue if additional differentiation is needed. Otherwise proceed directly to Step #12.
        1. One dip in Lithium Carbonate 0.05%, Aqueous (Step #2).
        2. Dip in two changes of 70% ethyl alcohol until greenish/blue white matter sharply contrasts with colorless gray matter.
    1. Rinse thoroughly in distilled water.
    2. Stain with Hematoxylin Stain, Harris Modified (Part 1201) for 1-5 minutes, depending on nuclear stain preference.
    3. Wash in running tap water for 3 minutes.
    4. Differentiate quickly in Acid Alcohol 1% (Part 10011); 3 dips.
    5. Wash well in running tap water.
    6. Blue in Solution B: Lithium Carbonate, Saturated Aqueous.
    7. Wash well in running tap water.
    8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Myelin Blue to blue/green
Nissl substance and nuclei Blue

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. 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. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 378.
    2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 206-211.
    3. Klüver, Heinrich and Elizabeth Barrera. “A Method for the Combined Staining of Cells and Fibers in the Nervous System.” Journal of Neuropathology and Experimental Neurology 12.4 (1953): 400-403.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining hamster liver.
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 Leptospira, Animal Control Slides are for the positive histochemical staining of Leptospira sp, a corkscrew shaped bacterium readily visualized with silver staining techniques.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. 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. Ingredient D: Hydroquinone, Powder                0.5 gm (or one rounded scoop with reusable mini sampling spoon)
        2. Distilled Water                 25 ml
    1. Prepare fresh Reducing Solution by combining in order listed.
        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.
    2. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Note #3
    2. 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. Place slides in a plastic Coplin jar (Part 5184) with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave 1 minute at 70°C.
    1. Rinse well in several changes of distilled water.
    2. 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. Place slides in a plastic Coplin jar with Solution B: Silver Nitrate 1%, Aqueous. Microwave 1 minute at 70°C.
        2. Remove from microwave, cover and let sit for 1 minute.
    1. Rinse well in several changes of distilled water.
        1. Excessive rinsing may cause nuclei to pick up silver.
    2. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
    3. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
    4. 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. 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. Place slides in a plastic Coplin jar with Reducing Solution. Microwave 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. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Leptospira Dark brown to black
Background Golden brown

 

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    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. 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 liver or spleen.
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 Control Slides are for the positive histochemical staining of ferric iron deposits in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Acid clean glassware prior to use to avoid residual iron staining.
        1. See Procedure Note #1.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

 

RESULTS:

Ferric iron deposits Bright blue
Nuclei Red
Cytoplasm Pink

 

PROCEDURE NOTES:

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

 

REFERENCES:

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

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:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:

Newcomer Supply Histoplasma, Animal Control Slides are for the positive histochemical staining of fungal organisms in tissue sections.  The morphology of the organisms is consistent with Histoplasma sp.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well.
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

STAINING PROCEDURE:

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

        Microwave Modification: See Procedure Note #5

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash in running tap water for 5 minutes; rinse well in distilled water.
    4. 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. 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. Staining at room temperature will require longer incubation.
    1. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    2. Rinse in three to four changes of distilled water.
        1. Do not use tap water at this step.
    3. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    4. Rinse well in distilled water.
    5. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    6. Wash in running tap water for 5 minutes; rinse in distilled water.
    7. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
    8. 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:

Histoplasma Sharply outlined in black
Background Green

 

PROCEDURE NOTES:

    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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th ed. Chicago: ASCP Press, 2020. 221-226.
    2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic”. American Journal of Clinical Pathology 25 (1955): 975-979.
    3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
    4. Sheehan, Dezna and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
    5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  H&E 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 Hematoxylin and Eosin (H&E) Stain: Individual Stain Solution
Hematoxylin Stain, Harris Modified
                  OR
Hematoxylin Stain, Harris		 Part 1201
OR
Part 12013
Acid Alcohol 1% Part 10011
Lithium Carbonate, Saturated Aqueous
               OR
Scott Tap Water Substitute		 Part 12215
OR
Part 1380
Eosin Y Working Solution Part 1072

 

APPLICATION:

Newcomer Supply Hematoxylin & Eosin (H&E) Control Slides are for the clear demonstration of nuclei, epithelium, connective tissue and muscle, for documentation of high quality nuclear and cytoplasmic staining.

For quality control measures, use H&E Control Slides each day that H&E staining is performed and additionally after any H&E solution changes.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Stain with Hematoxylin Stain, Harris Modified or Hematoxylin Stain, Harris for 1 to 5 minutes, depending on nuclear stain preference.
    4. Wash well in three changes of tap water.
    5. Differentiate quickly in Acid Alcohol 1%.
        1. See Procedure Note #3.
    6. Rinse immediately in three changes of tap water.
    7. Blue slides in Lithium Carbonate, Saturated Aqueous or Scott Tap Water Substitute for 10 dips.
    8. Wash in three changes of tap water; rinse in distilled water.
    9. Drain excess water from slides; proceed to 70% alcohol for 10 dips.
    10. Counterstain in Eosin Y Working Solution for 30 seconds to 3 minutes, depending on counterstain preference.
    11. Dehydrate in two changes of 95% ethyl alcohol for 1 minute each and two changes of 100% ethyl alcohol, 10 dips each.   Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Nuclei Blue
Cytoplasm of epithelium, connective tissue, muscle Shades of pink

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. Differentiate to suit preference of nuclear stain intensity.
        1. Check microscopically for satisfactory nuclear staining.
        2. Nuclei should be distinct and background light to colorless.
    4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 118-120.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 143-144, 153-154.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining gastrointestinal tract biopsy.
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 Helicobacter Control Slides are for the positive histochemical staining of helical shaped Helicobacter bacterium, associated with stomach inflammation and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. 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. Ingredient D: Hydroquinone, Powder                5 gm (or one rounded scoop with reusable mini sampling spoon)
        2. Distilled Water                 25 ml
    1. Prepare fresh Reducing Solution by combining in order listed.
        1. Hydroquinone Solution (Step #5) 25 ml
        2. Solution C: Gum Mastic 2.5%, Alcoholic         15 ml
        3. Solution B: Silver Nitrate 1%, Aqueous 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.
    2. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Note #3
    2. 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. Place slides in a plastic Coplin jar with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
    1. Rinse well in several changes of distilled water.
    2. Place slides in preheated Solution B: Silver Nitrate 1%, Aqueous (Step #4) and incubate in a 60°C water bath for 15 minutes.
    1.         Microwave Modification:
        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. Rinse well in several changes of distilled water.
        1. Excessive rinsing may cause nuclei to pick up silver.
    1. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
    2. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
    3. 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. 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. 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. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Helicobacter Dark brown to black
Background Golden brown

 

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    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. 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:  Thiazine 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 Differential Stain, Monochromatic Method: Individual Stain Solution
Thiazine Stain Part 10522

 

APPLICATION:

Newcomer Supply Helicobacter, Artificial Control Slides are for the positive histochemical staining of helical shaped Helicobacter bacterium, associated with stomach inflammation and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.

Helicobacter pylori purchased from American Type Culture Collection is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
    3. Place slides in Thiazine Stain for 1-4 minutes depending upon staining intensity preference.
    4. Rinse slides quickly in distilled water; long enough to remove excess stain.
    5. Allow slides to air-dry in a vertical position.
    6. Dip dried slides in xylene and coverslip with compatible mounting medium.
        1. See Procedure Note #1.

 

RESULTS:

Helicobacter Dark blue
Collagen and muscle Blue
Nuclei Blue
Cytoplasm Varying shades of light blue

 

PROCEDURE NOTES:

    1. Elimination of dehydration steps is necessary to retain dark stain of the organism.
    2. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and coverslipping steps.

 

REFERENCES:

    1. Potvin, Carol. “A Modified Diff-Quik Stain for Helicobacter pylori in Gastrointestinal Biopsies.” Laboratory Medicine 25.6 (1994): 389-391.
    2. Skipper, Ray and Don DeStephano. “A Rapid Stain for Campylobacter pylori in Gastrointestinal Tissue Sections Using Diff-Quik.” The Journal of Histotechnology 12.4 (1989): 303-304.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal gastrointestinal tract.
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 Helicobacter, Animal Control Slides are for the positive histochemical staining of helical shaped Helicobacter bacterium, associated with stomach inflammation and implicated in the development of gastric malignancy, peptic ulcers, and chronic gastritis.  These control slides are not validated for immunohistochemistry (IHC) use.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. 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. Ingredient D: Hydroquinone, Powder                0.5 gm  (or one rounded scoop with reusable mini sampling spoon)
        2. Distilled Water                 25 ml
    1. Prepare fresh Reducing Solution by combining in order listed.
        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.
    2. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Note #3.
    2. 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. Place slides in a plastic Coplin jar with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
    1. Rinse well in several changes of distilled water.
    2. 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. 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. Rinse well in several changes of distilled water.
        1. Excessive rinsing may cause nuclei to pick up silver.
    1. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
    2. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
    3. 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. 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. 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. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Helicobacter Dark brown to black
Background Golden brown

           

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    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. 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 spleen.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  May-Grunwald Giemsa 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 May-Grunwald Giemsa Stain: Individual Stain Solution
Jenner Stock Stain Part 1210
Giemsa Stock Stain, Wolbach Part 1121
Alcohol, Methanol Anhydrous, ACS Part 12236
Acetic Acid 1%, Aqueous Part 10012

 

APPLICATION:

Newcomer Supply Giemsa Control Slides are for the positive histochemical and differential staining of hematopoietic tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Rinse in two changes of Alcohol, Methanol Anhydrous, ACS (Part 12236); 3 minutes each.
    4. Prepare fresh Working Jenner Stain Solution just prior to use; combine and mix well.
        1. Distilled Water       20 ml
        2. Jenner Stock Stain                       20 ml
    5. Place slides in fresh Working Jenner Stain Solution for 6 minutes.
    6. Prepare fresh Working Giemsa Stain Solution just prior to use; combine and mix well.
        1. Distilled Water       47 ml
        2. Giemsa Stock Stain, Wolbach         3 ml
    7. Place slides directly into fresh Working Giemsa Stain Solution for 45 minutes.
    8. Rinse quickly in distilled water.
    9. Differentiate each slide individually in Acetic Acid 1%, Aqueous (Part 10012); 6-10 dips.
        1. Check microscopically for well differentiated nuclei.
    10. Rinse in distilled water.
    11. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Nuclei Blue/violet
Cytoplasm Pink/rose to lighter blue shades

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. The color range of the stained cells may vary depending upon fixation and degree of differentiation.
    4. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Luna, Lee G. Manual of Histologic Staining Methods of the Armed Forces Institute of Pathology. 3rd ed. New York: Blakiston Division, McGraw-Hill, 1968. 121-122.
    2. Shapiro, Stanley H. and Hilda Laufer. “Observations on Fixation and Staining of Bone Marrow Biopsies.” The Journal of Histotechnology 11.3 (1988): 145-47.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 157.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining Aspergillus rat lung, positive staining Candida rat lung, and positive staining Histoplasma animal tissue.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

                                                                                        

APPLICATION:

Newcomer Supply Fungus, GMS, Multi-Tissue, Artificial Control Slides, use a variety of lung tissue sources for the positive histochemical staining of Aspergillus sp., Candida sp., and organisms exhibiting morphology consistent with Histoplasma sp. in separate tissue sections.

Aspergillus fumigatus and Candida albicans purchased from Remel Microbiology Products are used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well:
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash for 5 minutes in running tap water; rinse well in distilled water.
    1. 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. 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. Staining at room temperature will require longer incubation.
    2. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    3. Rinse in three to four changes of distilled water.
        1. Never use tap water at this step.
    4. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    5. Rinse well in distilled water.
    6. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    7. Wash in running tap water for 5 minutes; rinse in distilled water.
    8. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
    9. 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:

Aspergillus Sharply outlined in black
Candida Sharply outlined in black
Histoplasma Sharply outlined in black
Background Green

 

PROCEDURE NOTES:

    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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th Chicago: ASCP Press, 2020. 221-226.
    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.

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:  PAS/Light Green 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 Fungus, PAS/Light Green Stain Kit: Part 9122A Individual Stain Solution
Solution A: Periodic Acid 0.5%, Aqueous 250 ml Part 13308
Solution B: Schiff Reagent, McManus 250 ml Part 1371
Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous 250 ml Part 12203

 

Newcomer Supply Fungus, PAS, Candida, Artificial Control Slides are for the positive histochemical staining of Candida in tissue sections.

Candida albicans purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place in Solution A: Periodic Acid 0.5%, Aqueous for 5 minutes.
    4. Wash in three changes of tap water; rinse in distilled water.
    5. Drain slides of excess water and stain in Solution B: Schiff Reagent, McManus for 20 minutes.
    6. Wash gently in lukewarm tap water for 10 minutes, allowing pink color to develop.
    7. Counterstain in Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous for 5 seconds.
        1. See Procedure Note #3.
    8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Candida Red to magenta
Background Pale green

 

PROCEDURE NOTES:

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

 

REFERENCES:

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

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:  PAS/Light Green 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 Fungus, PAS/Light Green Stain Kit: Part 9122A Individual Stain Solution
Solution A: Periodic Acid 0.5%, Aqueous 250 ml Part 13308
Solution B: Schiff Reagent, McManus 250 ml Part 1371
Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous 250 ml Part 12203

 

APPLICATION:

Newcomer Supply Fungus, PAS, Aspergillus, Artificial Control Slides are for the positive histochemical staining of Aspergillus in tissue sections.

Aspergillus fumigatus purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place in Solution A: Periodic Acid 0.5%, Aqueous for 5 minutes.
    4. Wash in three changes of tap water; rinse in distilled water.
    5. Drain slides of excess water and stain in Solution B: Schiff Reagent, McManus for 20 minutes.
    6. Wash gently in lukewarm tap water for 10 minutes, allowing pink color to develop.
    7. Counterstain in Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous for 5 seconds.
        1. See Procedure Note #3.
    8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Aspergillus Red to magenta
Background Pale green

  

PROCEDURE NOTES:

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

 

REFERENCES:

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

 

Be ready to handle potentially infectious specimens. Dispose of forceps after use to lessen the risk of exposure to disease.  Sterile 4 1/2″ forceps are exceptionally economical and are available in individually wrapped sterile packets.

25 Forceps/Pack

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:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:

Newcomer Supply Fungus, GMS, Candida, Artificial Control Slides are for the positive histochemical staining of Candida in tissue sections.

Candida albicans purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well:
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash for 5 minutes in running tap water; rinse well in distilled water.
    4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
        1. Periodically remove control, rinse in warm distilled water, check microscopically for adequate silver impregnation. Fungi should be dark brown.
        2. If organisms are not sufficiently dark, return slides to warm silver solution. Recheck at 2-3 minute intervals until desired intensity is achieved.
        3. Staining at room temperature will require longer incubation.
    1. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    2. Rinse in three to four changes of distilled water.
        1. Never use tap water at this step.
    3. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    4. Rinse well in distilled water.
    5. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    6. Wash in running tap water for 5 minutes; rinse in distilled water.
    7. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
    8. 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:

Candida Sharply outlined in black
Background Green

 

PROCEDURE NOTES:

    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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th Chicago: ASCP Press, 2020. 221-226.
    2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic.” American Journal of Clinical Pathology 25 (1955): 975-979.
    3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
    4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
    5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining organ.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:

Newcomer Supply Fungus, GMS, Aspergillus Control Slides are for the positive histochemical staining of fungal organisms in tissue sections. The organism morphology is consistent with Aspergillus sp.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well:
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash for 5 minutes in running tap water; rinse well in distilled water.
    4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
        1. Periodically remove control, rinse in warm distilled water, check microscopically for adequate silver impregnation. Fungi should be dark brown.
        2. If organisms are not sufficiently dark, return slides to warm silver solution. Recheck at 2-3 minute intervals until desired intensity is achieved.
        3. Staining at room temperature will require longer incubation.
    1. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    2. Rinse in three to four changes of distilled water.
        1. Never use tap water at this step.
    3. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    4. Rinse well in distilled water.
    5. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    6. Wash in running tap water for 5 minutes; rinse in distilled water.
    7. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.  
    8. 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:

Aspergillus Sharply outlined in black
Background Green

 

PROCEDURE NOTES:

    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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th Chicago: ASCP Press, 2020. 221-226.
    2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic.” American Journal of Clinical Pathology 25 (1955): 975-979.
    3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
    4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
    5. Modifications developed by Newcomer Supply Laboratory.

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: Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:

Newcomer Supply Fungus, GMS, Aspergillus, Artificial Control Slides are for positive histochemical staining of Aspergillus in tissue sections.

Aspergillus fumigatus purchased from Remel Microbiology Products is used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well:
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash for 5 minutes in running tap water; rinse well in distilled water.
    4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°-60°C or at room temperature, for 12-18 minutes until sections appear paper-bag brown.
        1. Periodically remove control, rinse in warm distilled water, check microscopically for adequate silver impregnation.  Fungi should be dark brown.
        2. If organisms are not sufficiently dark, return slides to warm silver solution. Recheck at 2-3 minute intervals until desired intensity is achieved.
        3. Staining at room temperature will require longer incubation.
    5. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    1. Rinse in three to four changes of distilled water.
        1. Never use tap water at this step.
    2. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    3. Rinse well in distilled water.
    4. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    5. Wash in running tap water for 5 minutes; rinse in distilled water.
    6. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.  
    7. 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:

Aspergillus Sharply outlined in black
Background Green

  

PROCEDURE NOTES:

    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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th ed. Chicago: ASCP Press, 2020. 221-226.
    2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic.” American Journal of Clinical Pathology 25 (1955): 975-979.
    3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
    4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
    5. Modifications developed by Newcomer Supply Laboratory.

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:  AFB, Fite quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

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

 

APPLICATION:

Newcomer Supply Fite, Nocardia, Artificial Control Slides are for the positive histochemical staining of Nocardia sp.  in tissue sections.

Nocardia asteroides, purchased from Remel Microbiology Products, is used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Filter Solution B: Carbol Fuchsin Stain, Ziehl-Neelsen with high quality filter paper.
    3. Prepare Diluted Acid Alcohol Solution:
        1. Solution C: Acid Alcohol 1% 20 ml
        2. Distilled Water                 20 ml

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize slides in Solution A: Xylene/Peanut Oil, 2:1, two changes for 12 minutes each.
        1. See Procedure Note #1.
    2. Drain slides, wipe off excess oil and blot to opacity, removing residual oil.
        1. See Procedure Note #2.
    3. Stain in freshly filtered Solution B: Carbol Fuchsin Stain, Ziehl-Neelsen for 15 minutes at room temperature.
    4. Rinse well in distilled water.
    5. Differentiate slides individually in Diluted Acid Alcohol Solution (Step #3) until background is light pink; 10-20 dips.
    6. Quickly rinse in distilled water and check microscopically for correct differentiation.
    7. Rinse well in distilled water.
    8. Counterstain in Solution D: Light Green SF Yellowish 0.1%, Aqueous; 5-10 dips.
    9. Rinse in distilled water.
    10. Blot excess water from slide and air-dry or oven-dry completely.
    11. Dip dried slides in xylene and coverslip with a compatible mounting medium.

 

RESULTS:

Nocardia Red
Other tissue elements Green

 

PROCEDURE NOTES:

    1. Acid-fastness of Nocardia organisms is enhanced when the waxy capsule is protected by mixture of xylene/peanut oil and avoiding dehydrating solutions.
    2. It is important to blot well; residual oil may produce staining artifact.
    3. A small percentage of Nocardia sp. organisms may resist absorption of red carbol fuchsin stain and retain green counterstain due to growth phase of individual organisms.
    4. If using a xylene substitute, follow manufacturer’s recommendation for coverslipping step.

 

REFERENCES:

    1. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-instructional Text. 5th ed. Chicago: ASCP Press, 2020. 215-216.
    2. Fite, George, P.J. Cambre and M.H. Turner. “Procedure for Demonstrating Lepra Bacilli in Paraffin Sections”. Archives of Pathology 43 (1947). 624-625.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining skin.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Verhoeff-Van Gieson Elastic 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 Elastic, Verhoeff Stain Kit: Part 9116A/B Individual Stain Solution
Solution A: Hematoxylin 5%, Alcoholic 125/250 ml Part 11623
Solution B: Ferric Chloride 10%, Aqueous 125/250 ml Part 10856
Solution C: Iodine, Lugol’s, Aqueous 75/150 ml Part 12092
Solution D: Sodium Thiosulfate 5%, Aqueous 250/500 ml  Part 1389
Solution E: Van Gieson Stain 250/500 ml Part 1404

 

APPLICATION: 

Newcomer Supply Elastic, Skin Control Slides are for the positive histochemical staining of elastic fibers in skin.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Prepare fresh Verhoeff Working Solution by combining in the exact order listed, mixing well after each addition. Save for Step #5.
        1. Solution A: Hematoxylin 5%, Alcoholic                       20 ml
        2. Solution B: Ferric Chloride 10%, Aqueous         8 ml
        3. Solution C: Iodine, Lugol’s, Aqueous                         8 ml
    3. Prepare fresh Ferric Chloride 2%, Aqueous Solution for Step #7.
        1. Solution B: Ferric Chloride 10%, Aqueous       10 ml
        2. Distilled water       40 ml

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    2. Stain in fresh Verhoeff Working Solution (Step #2) for 20 minutes.
        1. Discard solution after successful differentiation in Step #7.
    3. Rinse in several changes of tap water.
    4. Differentiate each slide individually with agitation in fresh Ferric Chloride 2%, Aqueous Solution (Step #3); approximately 5-10 dips.
    5. Check differentiation: rinse well in tap water and check microscopically for black elastic staining with gray background.
        1. If necessary, return to Ferric Chloride 2%, Aqueous Solution until desired elastic differentiation is achieved.
        2. See Procedure Notes #3 and #4.
    6. Wash well in tap water.
    7. Place in Solution D: Sodium Thiosulfate 5%, Aqueous for 1 minute.
    8. Wash well in running tap water for 5 minutes.
    9. Counterstain in Solution E: Van Gieson Stain for 3 to 5 minutes.
    10. 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:

Elastic fibers/tissue/nuclei Blue-black to black
Collagen Red
Other tissue elements Yellow

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. If background is colorless, the section is over-differentiated.
        1. Return to Step #5, restain and reduce differentiation dips.
    4. Differentiation can vary between slides depending on amount of elastic tissue present in sections.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 167-169.
    2. Mallory, Frank Burr and James Homer Wright. Pathological Technique. 7th ed. Philadelphia, PA: W.B. Saunders Company, 1918. 118-119.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining aorta.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Verhoeff-Van Gieson Elastic 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 Elastic, Verhoeff Stain Kit: Part 9116A/B Individual Stain Solution
Solution A: Hematoxylin 5%, Alcoholic 125/250 ml Part 11623
Solution B: Ferric Chloride 10%, Aqueous 125/250 ml Part 10856
Solution C: Iodine, Lugol’s, Aqueous 75/150 ml Part 12092
Solution D: Sodium Thiosulfate 5%, Aqueous 250/500 ml Part 1389
Solution E: Van Gieson Stain 250/500 ml Part 1404

 

APPLICATION:

Newcomer Supply Elastic, Aorta Control Slides are for the positive histochemical staining of elastic fibers in artery.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Prepare fresh Verhoeff Working Solution by combining in the exact order listed, mixing well after each addition. Save for Step #5.
        1. Solution A: Hematoxylin 5%, Alcoholic                       20 ml
        2. Solution B: Ferric Chloride 10%, Aqueous         8 ml
        3. Solution C: Iodine, Lugol’s, Aqueous                         8 ml
    3. Prepare fresh Ferric Chloride 2%, Aqueous Solution for Step #7.
        1. Solution B: Ferric Chloride 10%, Aqueous       10 ml
        2. Distilled water       40 ml

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    2. Stain in fresh Verhoeff Working Solution (Step #2) for 20 minutes.
        1. Discard solution after successful differentiation in Step #7.
    3. Rinse in several changes of tap water.
    4. Differentiate each slide individually with agitation in fresh Ferric Chloride 2%, Aqueous Solution (Step #3); approximately 5-10 dips.
    5. Check differentiation: rinse well in tap water and check microscopically for black elastic staining with gray background.
        1. If necessary, return to Ferric Chloride 2%, Aqueous Solution until desired elastic differentiation is achieved.
        2. See Procedure Notes #3 and #4.
    6. Wash well in tap water.
    7. Place in Solution D: Sodium Thiosulfate 5%, Aqueous for 1 minute.
    8. Wash well in running tap water for 5 minutes.
    9. Counterstain in Solution E: Van Gieson Stain for 3 to 5 minutes.
    10. 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:

Elastic fibers/tissue/nuclei Blue-black to black
Collagen Red
Other tissue elements Yellow

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. If background is colorless, the section is over-differentiated.
        1. Return to Step #5, restain and reduce differentiation dips.
    4. Differentiation can vary between slides depending on amount of elastic present in sections.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 167-169.
    2. Mallory, Frank Burr and James Homer Wright. Pathological Technique. 7th ed. Philadelphia, PA: W.B. Saunders Company, 1918. 118-119.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining lung.
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 Cryptococcus Control Slides are for the positive histochemical staining of fungal organisms.  The morphology of the organism is consistent with Cryptococcus sp.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Prepare fresh Weigert Iron Hematoxylin Working Solution directly before use; combine and mix well:
        1. Solution A: Ferric Chloride, Aqueous   20 ml
        2. Solution B: Hematoxylin 1%, Alcoholic             20 ml
    4. Stain in 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. Solution C: Mucicarmine Stock Stain, Mayer   10 ml  
        2. Tap Water (do not use distilled water)   30 ml    
    7. Stain slides 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. Place slides in a plastic Coplin jar (Part 5184) containing fresh Mayer Mucicarmine Working Solution. Microwave for 10 minutes at 70°C.
    1. Rinse in several changes of tap water.
    2. Counterstain in Solution D: Metanil Yellow Stain, Aqueous for 30 seconds to 1 minute.
    3. Dehydrate quickly through 95% and 100% ethyl alcohols. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Capsule of Cryptococcus Deep rose to red
Nuclei Black
Other tissue elements Yellow

 

PROCEDURE NOTES:

    1. Drain staining 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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th ed. Chicago: ASCP Press, 2020. 149-151.
    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. 161-162.
    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.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining animal liver.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Rhodanine 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 Copper, Rhodanine Stain Kit: Part 9113A Individual Stain Solution
Solution A: Rhodanine Stock Stain 0.2%, Alcoholic 50 ml Part 10531
Solution B: Hematoxylin Stain, Mayer Modified 250 ml Part 1202
Solution C: Sodium Borate 0.5%, Aqueous 500 ml Part 13824

 

APPLICATION:

Newcomer Supply Copper, Animal Control Slides are for the positive histochemical detection of copper in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Prepare Working Rhodanine Solution; combine and mix well.
        1. Shake Solution A: Rhodanine Stock Stain 0.2%, Alcoholic well before each use.
        2. Solution A: Rhodanine Stock Stain 0.2%, Alcoholic 3 ml
        3. Distilled Water    47 ml

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    2. Stain in Working Rhodanine Solution (Step #2) at 60°C for 1-2 hours or at 37°C for 18 hours.

        Microwave Modification:  See Procedure Note #3.

        1. Place slides in a plastic Coplin jar containing Working Rhodanine Solution. Microwave for 6 minutes at 70°C.
    1. At end of incubation (for both oven and microwave), to avoid unwanted slide precipitate, pour off warm Working Rhodanine Solution into a second Coplin jar; reserve and set aside.
    2. Rinse slides well in several changes of distilled water.
    3. Check positive control slide microscopically to determine adequate copper/reddish brown development.
        1. Return slides to reserved Working Rhodanine Solution if additional incubation is required.
    4. Prepare dilute Mayer Hematoxylin Stain Solution directly before use; combine and mix well:
        1. Solution B: Hematoxylin Stain, Mayer Modified      20 ml
        2. Distilled Water     20 ml
    5. Stain in dilute Mayer Hematoxylin Stain Solution for 10 minutes.
    6. Rinse in distilled water.
    7. Rinse in Solution C: Sodium Borate 0.5%, Aqueous; 2-3 quick dips.
    8. Rinse well in distilled water.
    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:

Copper Copper/reddish brown
Nuclei Light blue

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. 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. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 251.
    2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 258-260.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 230.
    4. Modifications developed by Newcomer Supply.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine and positive iron staining animal spleen.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Müller-Mowry Colloidal Iron 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 Colloidal Iron, Müller-Mowry Stain Kit: Part 9110A Individual Stain Solution
Solution A: Acetic Acid 12%, Aqueous 1000 ml
Solution B: Colloidal Iron Stock 125 ml Part 10365
Solution C: Acetic Acid, Glacial, ACS 50 ml Part 10010
Solution D: Potassium Ferrocyanide 2%, Aqueous 125 ml
Solution E: Hydrochloric Acid 2%, Aqueous 125 ml
Solution F: Van Gieson Stain 250 ml Part 1404

 

APPLICATION:

Newcomer Supply Colloidal Iron, Multi-Tissue Control Slides, use a combination of tissue sources for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) and stromal (mesenchymal) mucin in small intestine and ferric iron in spleen.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Acid clean glassware prior to use to avoid residual iron staining.
        1. See Procedure Note #1.
    3. Prepare Colloidal Iron Working Solution; combine and mix well.
        1. Solution B: Colloidal Iron Stock                          20 ml
        2. Solution C: Acetic Acid, Glacial ACS                    5 ml
        3. Distilled Water                                                    15 ml

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #2 and #3.
    2. Place in Solution A: Acetic Acid 12%, Aqueous for 30 seconds.
    3. Drain Slides. Do not rinse.
    4. Place in Colloidal Iron Working Solution (Step #3) for 30 minutes.
    5. Rinse in three changes of Solution A: Acetic Acid 12%, Aqueous; 3 minutes each.
    6. Prepare fresh Ferrocyanide-Hydrochloric Acid Solution directly before use; combine and mix well.
        1. Solution D: Potassium Ferrocyanide 2%, Aqueous 20 ml
        2. Solution E: Hydrochloric Acid 2%, Aqueous        20 ml
    7. Place in Ferrocyanide-Hydrochloric Acid Solution for 15 minutes.
    8. Wash in running tap water for 1-5 minutes.
    9. Counterstain in Solution F: Van Gieson Stain for 3-5 minutes.
        1. Proceed directly to dehydration step without rinsing.
    10. Dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Blue
Stromal mucin Blue
Ferric iron deposits Bright blue
Collagen Red
Muscle and cytoplasm Yellow

 

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    2. Drain slides after each step to prevent solution carry over.
    3. Do not allow sections to dry out at any point during procedure.
    4. Nuclear Fast Red Stain, Kernechtrot (Part 1255) is an alternative counterstain.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 175-176.
    2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 151-153.
    3. Rekhtman, Natasha and Justin Bishop. Quick Reference Handbook for Surgical Pathologists. Berlin: Springer, 2011. 69.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Müller-Mowry Colloidal Iron 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 Colloidal Iron, Müller-Mowry Stain Kit: Part 9110A Individual Stain Solution
Solution A: Acetic Acid 12%, Aqueous 1000 ml
Solution B: Colloidal Iron Stock 125 ml Part 10365
Solution C: Acetic Acid, Glacial, ACS 50 ml Part 10010
Solution D: Potassium Ferrocyanide 2%, Aqueous 125 ml
Solution E: Hydrochloric Acid 2%, Aqueous 125 ml
Solution F: Van Gieson Stain 250 ml Part 1404

 

APPLICATION:

Newcomer Supply Colloidal Iron Control Slides are for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) and stromal (mesenchymal) mucin in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Acid clean glassware prior to use to avoid residual iron staining.
        1. See Procedure Note #1.
    3. Prepare Colloidal Iron Working Solution; combine and mix well.
        1. Solution B: Colloidal Iron Stock                          20 ml
        2. Solution C: Acetic Acid, Glacial ACS                    5 ml
        3. Distilled Water                                                    15 ml

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #2 and #3.
    2. Place in Solution A: Acetic Acid 12%, Aqueous for 30 seconds.
    3. Drain Slides. Do not rinse.
    4. Place in Colloidal Iron Working Solution (Step #3) for 30 minutes.
    5. Rinse in three changes of Solution A: Acetic Acid 12%, Aqueous; 3 minutes each.
    6. Prepare fresh Ferrocyanide-Hydrochloric Acid Solution directly before use; combine and mix well.
        1. Solution D: Potassium Ferrocyanide 2%, Aqueous 20 ml
        2. Solution E: Hydrochloric Acid 2%, Aqueous        20 ml
    7. Place in Ferrocyanide-Hydrochloric Acid Solution for 15 minutes.
    8. Wash in running tap water for 1-5 minutes.
    9. Counterstain in Solution F: Van Gieson Stain for 3-5 minutes.
        1. Proceed directly to dehydration step without rinsing.
    10. Dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Blue
Stromal mucin Blue
Collagen Red
Muscle and cytoplasm Yellow

 

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    2. Drain slides after each step to prevent solution carry over.
    3. Do not allow sections to dry out at any point during procedure.
    4. Nuclear Fast Red Stain, Kernechtrot (Part 1255) is an alternative counterstain.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 175-176.
    2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 151-153.
    3. Rekhtman, Natasha and Justin Bishop. Quick Reference Handbook for Surgical Pathologists. Berlin: Springer, 2011. 69.
    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:  Steiner-Steiner Modified 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 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 Cat Scratch Artificial Control Slides are for the positive histochemical staining and qualitative identification of proteins of Bartonella sp., the causative bacterial agent of Cat Scratch disease.

Bartonella henselae purchased from American Type Culture Collection is used to produce the positive control tissue.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. 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. Ingredient D: Hydroquinone, Powder    0.5 gm            (or one rounded scoop with reusable mini sampling spoon)
        2. Distilled Water          25 ml
    1. Prepare fresh Reducing Solution by combining in order listed.
        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.
    2. Do not preheat solutions if using Microwave Modifications.

 

STAINING PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Note #3 
    2. 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. Place slides in a plastic Coplin jar with Solution A: Uranyl Nitrate 1%, Aqueous. Microwave for 1 minute at 70°C.
    1. Rinse well in several changes of distilled water.
    2. 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. 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. Rinse well in several changes of distilled water.
        1. Excessive rinsing may cause nuclei to pick up silver.
    2. Dip 5 times in two changes of fresh 95% and 100% ethyl alcohols.
    3. Place in Solution C: Gum Mastic 2.5%, Alcoholic for 3 minutes.
    4. 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. 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. 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. Directly dehydrate in two changes of 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Bartonella  Dark brown to black
Background Golden brown

 

PROCEDURE NOTES:

    1. Acid clean all glassware/plasticware (Part 12086) and rinse thoroughly in several changes of distilled water.
    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. 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 placenta.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Von Kossa Calcium 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 Von Kossa Calcium Stain: Individual Stain Solution
Silver Nitrate 5%, Aqueous Part 13805
Sodium Thiosulfate 5%, Aqueous Part 1389
Nuclear Fast Red Stain, Kernechtrot Part 1255

 

APPLICATION:

Newcomer Supply Calcium Control Slides are for the positive histochemical staining of calcium or calcium salts in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to you laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #3 and #4.
    2. Place slides in Silver Nitrate 5%, Aqueous according to the following timings and conditions.
        1. Direct sunlight or ultraviolet light for 10-30 minutes.
        2. In front of a 60-100 watt light bulb for 1 hour or longer.
        3. See Procedure Note #5.
    3. Check slides periodically and remove from light source when control slide shows black-brown deposits macroscopically.
    4. Rinse well in several changes of distilled water.
    5. Place slides in Sodium Thiosulfate 5%, Aqueous for 2 minutes.
    6. Rinse well in several changes of distilled water.
    7. Counterstain in Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
        1. Shake solution well before use; do not filter.    
    8. Rinse well in distilled water.
        1. See Procedure Note #6.
    9. Dehydrate in two changes each of 95% and 100% ethyl alcohol; 10 dips each. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Calcium salts Black to brown/black
Nuclei Red
Cytoplasm Light pink

 

PROCEDURE NOTES:

    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 (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. Direct sunlight is the preferred method. If procedure is in minimal sunlight increased incubation time will be necessary.
    6. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
    7. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 269-270.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 226-227.
    3. 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:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml  Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:                                           

Newcomer Supply Blastomyces, Animal Control Slides are for the positive histochemical staining of fungal organisms. The morphology of the organisms is consistent with Blastomyces sp.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well.
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

STAINING PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash in running tap water for 5 minutes; rinse well in distilled water.
    4. 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. 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. Staining at room temperature will require longer incubation.
    1. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    1. Rinse in three to four changes of distilled water.
        1. Do not use tap water at this step.
    1. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    2. Rinse well in distilled water.
    3. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    4. Wash in running tap water for 5 minutes; rinse in distilled water.
    5. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
    6. 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:

Blastomyces Crisp black cell walls & visible internal structures
Mucin Taupe to dark gray
Background Green

 

PROCEDURE NOTES:

      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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th ed. Chicago: ASCP Press, 2020. 221-226.
      2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic”. American Journal of Clinical Pathology 25 (1955): 975-979.
      3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
      4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
      5. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining lung.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Grocott 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 Fungus, Grocott Methenamine Silver (GMS) Stain Kit: Part 9121A/B  Individual Stain Solution
Solution A: Chromic Acid 5%, Aqueous 250/500 ml Part 10341
Solution B: Sodium Bisulfite 1%, Aqueous 250/500 ml Part 13821
Solution C: Silver Nitrate 125/250 ml Part 1142
Solution D: Methenamine Borate 125/250 ml Part 1142
Solution E: Gold Chloride 0.1%, Aqueous 250/500 ml Part 11285
Solution F: Sodium Thiosulfate 2%, Aqueous 250/500 ml Part 13888
Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous 250/500 ml Part 12202

 

APPLICATION:                                           

Newcomer Supply Blastomyces Control Slides are for the positive histochemical staining of fungal organisms. The morphology of the organisms is consistent with Blastomyces sp.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well.
        1. Solution C: Silver Nitrate 20 ml
        2. Solution D: Methenamine Borate 20 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath approximately 20 to 30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat if using Microwave Modification; Step 11.

 

STAINING PROCEDURE:

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

        Microwave Modification: See Procedure Note #5.

        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. Wash well in running tap water; rinse in distilled water.
    2. Place in Solution B: Sodium Bisulfite 1%, Aqueous for 1 minute.
    3. Wash in running tap water for 5 minutes; rinse well in distilled water.
    4. 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. 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. Staining at room temperature will require longer incubation.
    1. Microwave Modification:
        1. Incubate slides in a plastic Coplin jar containing 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 3-5 minute intervals.
    1. Rinse in three to four changes of distilled water.
        1. Do not use tap water at this step.
    1. Tone in Solution E: Gold Chloride 0.1%, Aqueous until sections turn gray; 10-30 seconds.
    2. Rinse well in distilled water.
    3. Remove unreduced silver in Solution F: Sodium Thiosulfate 2%, Aqueous for 2 minutes.
    4. Wash in running tap water for 5 minutes; rinse in distilled water.
    5. Counterstain in Solution G: Light Green SF Yellowish Stain 0.2%, Aqueous for 2 minutes.
    6. 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:

Blastomyces Crisp black cell walls & visible internal structures
Mucin Taupe to dark gray
Background Green

 

PROCEDURE NOTES:

      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. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-Instructional Text. 5th ed. Chicago: ASCP Press, 2020. 221-226.
      2. Grocott, R G, “A Stain for Fungi in Tissue Sections and Smears using Gomori Methenamine Silver Nitrate Technic”. American Journal of Clinical Pathology 25 (1955): 975-979.
      3. Koski, John. “Silver Methenamine Borate (SMB): Cost Reduction with Technical Improvement in Silver Nitrate-Gold Chloride Impregnations.” The Journal of Histotechnology 4.3 (1981): 115-119.
      4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 245-246.
      5. Modifications developed by Newcomer Supply Laboratory.

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:  Bile Stain, Hall’s Method 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 Bile Stain, Hall’s Method: Individual Stain Solution
Fouchet Reagent Part 1095
Van Gieson Stain Part 1404

 

APPLICATION:                        

Newcomer Supply Bile Control Slides are for the positive histochemical staining of bile (bilirubin) substances in tissue sections and to distinguish bile pigments from other tissue pigments.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Filter Fouchet Reagent with high quality filter paper prior to use.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

 

RESULTS:

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

 

PROCEDURE NOTES:

    1. Drain staining 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. 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 brain.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 8 microns on Superfrost™ Plus slides.
Quality Control Stain:  Bielschowsky 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 Bielschowsky, Lester King Modified Stain Kit:  Part 9154A Individual Stain Solution
Solution A: Silver Nitrate 20%, Aqueous 250 ml Part 13807
Solution B: Ammonium Hydroxide 28-30%, ACS 100 ml Part 1006
Solution C: Developer 25 ml
Solution D: Sodium Thiosulfate 5%, Aqueous 250 ml Part 1389

 

APPLICATION:

Newcomer Supply Bielschowsky Control Slides are for the positive histochemical staining of nerve fibers, neurofibrils/tangles, senile plaques and axons, instrumental in the diagnosis of Alzheimer’s disease and other neurological disorders.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Preheat Coplin jar of Solution A: Silver Nitrate 20%, Aqueous in water bath to 37°C.
    4. Preheat two Coplin jars of distilled water in 37°C water bath.
        1. Save for slide rinsing/holding in Steps #7 and #10.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #3 and #4.
    2. Place slides in preheated Solution A: Silver Nitrate 20%, Aqueous (Step #3) for 15 minutes.
        1. See Procedure Note #5.
    3. Remove slides from Solution A: Silver Nitrate 20%, Aqueous and hold in 1st reserved Coplin jar of preheated distilled water.
        1. Save Silver Nitrate 20%, Aqueous for Step #8.
    4. Add Solution B: Ammonium Hydroxide 28-30%, ACS drop by drop in saved Silver Nitrate 20%, Aqueous swirling until precipitate disappears. Do not go past this point. 
        1. Approximately 10 ml of Ammonium Hydroxide 28-30%, ACS will be required.
    5. Place slides back in Silver Nitrate Solution with added Ammonium Hydroxide in water bath at 37°C for 10 minutes.
    6. Remove slides and hold in 2nd reserved Coplin jar of distilled water.
        1. Save Ammoniacal Silver Solution for Step #11.
    7. Add 1 drop of Solution C: Developer to the saved Ammoniacal Silver Solution with swirling motion.
    8. Return slides to Ammoniacal Silver Solution with added Developer, in 37°C water bath for 5-15 minutes; average time of 6 minutes.
        1. Check slides microscopically at 3 minutes for development of neurons to dark brown.
        2. Then check at 1 minute intervals to avoid over-development.
    9. Rinse thoroughly in distilled water for 5 minutes.
    10. Place in Solution D: Sodium Thiosulfate 5%, Aqueous; 5 minutes.
    11. Rinse thoroughly in tap 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:

Senile plaques, neurofibrils/tangles Dark brown to black
Neurons Dark brown
White and gray matter Yellowish brown
Nerve fibers, axons Brown to black

 

PROCEDURE NOTES:

    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. A maximum of 8 slides per 40 ml of Solution A: Silver Nitrate 20%, Aqueous is recommended for proper silver development.
    6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-instructional Text. 4th ed. Chicago: ASCP Press, 2015. 196-199.
    2. King, Lester. “The Impregnation of Neurofibrils”. Yale Journal of Biology and Medicine 14.1 (1941). 59-68.
    3. 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:  Jones Basement Membrane 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 Jones Basement Membrane Stain Kit Part 9167A Individual Stain Solution
Solution A: Methenamine 3%, Aqueous 250 ml Part 12239
Solution B: Silver Nitrate 5%, Aqueous 50 ml Part 13805
Solution C: Sodium Borate 5%, Aqueous 50 ml Part 13826
Solution D: Periodic Acid 0.5%, Aqueous 250 ml Part 13308
Solution E: Gold Chloride 0.25%, Aqueous 250 ml Part 11287
Solution F: Sodium Thiosulfate 2.5%, Aqueous 250 ml Part 13889
Solution G: Light Green SF Yellowish Stain 0.1%, Aqueous 250 ml Part 12203

 

APPLICATION:

Newcomer Supply Basement Membrane Control Slides are for positive histochemical staining of basement membranes in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. All glassware/plasticware must be acid cleaned prior to use.
        1. See Procedure Notes #1 and #2.
    3. Prepare Silver-Methenamine Working Solution and mix well:
        1. Solution A: Methenamine 3%, Aqueous      40 ml
        2. Solution B: Silver Nitrate 5%, Aqueous 2 ml
        3. Solution C: Sodium Borate 5%, Aqueous 4 ml
    4. Preheat Silver-Methenamine Working Solution to 45°-60°C in a water bath 20-30 minutes before use.
        1. Maintain solution between 45°-60°C to minimize precipitate.
        2. Do not preheat solution if using Microwave Modification.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #3 and #4.
    2. Place in Solution D: Periodic Acid 0.5%, Aqueous for 15 minutes.
    3. Wash in tap water for 5 minutes; rinse in distilled water.
    4. Incubate slides in preheated Silver-Methenamine Working Solution (Step #4) at 45°-60°C oven or at room temperature for 12-18 minutes until sections appear paper-bag brown.
    5. Periodically remove control; rinse in warm distilled water, check microscopically for adequate silver impregnation. Basement membranes should be dark brown. If tissue structures are not sufficiently dark, return slides to warm silver solution.  Recheck at 2-3 minute intervals until desired intensity is achieved.
        1. Staining at room temperature will require longer incubation.
    6. Microwave Modification: See Procedure Note #5.
        1. Place sides in a plastic Coplin jar with prepared Silver-Methenamine Working Solution (Step #3). Microwave for 3 minutes at 70°C.
        2. Check microscopically for adequate development.
        3. If additional incubation is required, return slides to heated silver solution and recheck at regular intervals.
    1. Rinse in three changes of distilled water.
    2. Tone in Solution E: Gold Chloride 0.25%, Aqueous for 1 minute.
    3. Rinse well in three changes of distilled water.
    4. Place in Solution F: Sodium Thiosulfate 2.5%, Aqueous; 2 minutes.
    5. Wash in tap water for 5 minutes; rinse in distilled water.
    6. Counterstain in Solution G: Light Green SF Yellowish Stain 0.1%, Aqueous for 1 minute.
    7. Quickly rinse slides in two changes of distilled water.
    8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Kidney glomerular basement membranes Black
Intra-glomerular deposits Black
Reticular Fibers Black
Nuclei Outlined in black
Background Light Green

 

PROCEDURE NOTES:

    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. Jones, David B. “Nephrotic Glomerulonephritis,” American Journal of Pathology 33.2 (1957): 313–329.
    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. 97-99.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 187-188.
    4. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Alcian Blue pH 2.5 quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

With Alcian Blue, 1%, pH 2.5 Stain Kit: Part 9102A/B Individual Stain Solution
Solution A: Acetic Acid 3%, Aqueous 250/500 ml Part 10017
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250/500 ml Part 1003
Solution C: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

 

APPLICATION:

Newcomer Supply Alcian Blue pH 2.5, Goblet Cell Control Slides are for the positive histochemical staining of goblet cells as well as other acid epithelial mucins (sialomucin, sulfomucin).  Goblet cells normally line the small intestine but are an abnormal finding in Barrett’s esophagus.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
    4. Move slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous. Stain for 30 minutes at room temperature or for 15 minutes in a 37°C water bath.
        1. See Procedure Note #3.
    5. Wash in running tap water for 10 minutes; rinse in distilled water.
    6. Counterstain in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
        1. Shake solution well before use; do not filter.
    7. Rinse well in distilled water.
        1. See Procedure Note #4
    8. Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Goblet cells  Bright blue
Acid epithelial mucin Blue
Nuclei Pink-red
Cytoplasm Pale pink

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. A brief dip in Solution A: Acetic Acid 3%, Aqueous from Step #3 can be added before Step #5 to remove excess Alcian Blue Stain.
    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. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 145-148.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-175.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

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

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

 

CONTROL SLIDE VALIDATION:

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

 

APPLICATION:                                           

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

 

PRESTAINING PREPARATION:

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

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

 

RESULTS:

Melanin and argentaffin granules Black
Nuclei Pink-red

 

PROCEDURE NOTES:

    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. Wash well after Nuclear Fast Red Stain, Kernechtrot to avoid cloudiness in dehydration steps.
    6. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES: 

    1. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 286-287.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 276-277.
    3. 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 8 microns on Superfrost™ Plus slides.
Quality Control Stain:  Bennhold Congo Red 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 Amyloid, Bennhold Congo Red Stain Kit: Part 9103A Individual Stain Solution
Solution A: Congo Red Stain 1%, Aqueous 250 ml Part 1038
Solution B: Alkaline Alcohol 250 ml Part 1038
Solution C: Hematoxylin Stain, Mayer Modified 250 ml Part 1202

   

APPLICATION:

Newcomer Supply Amyloid, Animal Control Slides are for the positive histochemical staining of extraneous protein deposits in amyloidosis.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    2. Place slides in Solution A: Congo Red Stain 1%, Aqueous; 1 hour.

        Microwave Modification: See Procedure Note #3.

        1. Place slides in a plastic Coplin jar (Part 5184) containing Solution A: Congo Red Stain 1%, Aqueous. Microwave for 3 minutes at 70°C.
    1. Rinse in two to three changes of tap water; rinse in distilled water.
    2. Differentiate in Solution B: Alkaline Alcohol, 5 to 30 seconds, agitating constantly until slide background is cleared of Solution A: Congo Red Stain 1%, Aqueous.
    3. Rinse in two to three changes of tap water; rinse in distilled water.
    4. Counterstain with Solution C: Hematoxylin Stain, Mayer Modified, 3 to 5 minutes.
    5. Wash in running tap water for 5 to 10 minutes.
    6. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Light Field Microscopy:
Amyloid Pink to red
Nuclei Blue
Polarized Light:
Amyloid fluorescence Green, red, orange and yellow

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. 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. Sections are cut at 8 microns to provide more intense staining and allow smaller amyloid deposits to be identified.
    5. If using a xylene substitute, follow manufacturer’s recommendation for deparaffinization and clearing steps.

 

REFERENCES:

    1. Buxbaum, Joel N, David S. Eisenberg, Marcus Fändrich, Ellen D. McPhail, Giampaolo Merlini, Maria J. M. Saraiva, Yoshiki Sekijima and Per Westermark, “Amyloid nomenclature 2024: update, novel proteins, and recommendations by the International Society of Amyloidosis (ISA) Nomenclature Committee,” Amyloid 31, no. 4 (Sept 2024): 249-256, DOI: 10.1080/13506129.2024.2405948.
    2. Howie, Alexander J and Owen-Casey Mared P., “Systematic review of accuracy of reporting of Congo red-stained amyloid in 2010-2020 compared with earlier.” Annuls of Medicine 54, no. 1 (Dec 2022): 2511-2516, DOI: 10.1080/07853890.2022.2123558.
    3. Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts. Gaitheresburg, MD: American Histolabs, 1992. 366-367.
    4. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 177-178.
    5. Suvarna, S. Kim., Christopher Layton, and John D. Bancroft. Bancroft’s Theory and Practice of Histological Techniques. 8th ed. Oxford: Elsevier, 2019. 231-243.
    6. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining Barrett’s esophagus.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Alcian Blue pH 2.5 quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

With Alcian Blue 1%, pH 2.5 Stain Kit: Part 9102A/B Individual Stain Solution
Solution A: Acetic Acid 3%, Aqueous 250/500 ml Part 10017
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250/500 ml Part 1003
Solution C: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

 

APPLICATION:

Newcomer Supply Alcian Blue pH 2.5, Barrett’s Esophagus Control Slides are for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) as well as stromal (mesenchymal) mucin and  demonstrates the presence of columnar epithelium with goblet cells and stratified squamous epithelium in esophageal biopsy.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
    4. Move slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous. Stain for 30 minutes at room temperature or 15 minutes in a 37°C water bath.
        1. See Procedure Note #3.
    5. Wash in running tap water for 10 minutes; rinse in distilled water.
    6. Counterstain in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
        1. Shake solution well before use; do not filter.
    7. Rinse well in distilled water.
        1. See Procedure Note #4
    8. Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Blue
Stromal (mesenchymal) mucin Blue
Goblet cells Pale blue
Nuclei Pink-red
Cytoplasm Pale pink

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. A brief dip in Solution A: Acetic Acid 3%, Aqueous from Step #3 can be added before Step #5 to remove excess Alcian Blue Stain.
    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. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 145-148.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-175.
    3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining umbilical cord and positive staining small intestine.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Alcian Blue pH 2.5 quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

With Alcian Blue 1%, pH 2.5 Stain Kit: Part 9102A/B Individual Stain Solution
Solution A: Acetic Acid 3%, Aqueous 250/500 ml Part 10017
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250/500 ml Part 1003
Solution C: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

 

APPLICATION:

Newcomer Supply Alcian Blue pH 2.5, Multi-Tissue Control Slides use a combination of tissue sources for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) as well as stromal (mesenchymal) mucin.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
    4. Move slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous. Stain for 30 minutes at room temperature or for 15 minutes in a 37°C water bath.
        1. See Procedure Note #3.
    5. Wash in running tap water for 10 minutes; rinse in distilled water.
    6. Counterstain in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
        1. Shake solution well before use; do not filter.
    7. Rinse well in distilled water.
        1. See Procedure Note #4.
    8. Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Blue
Stromal (mesenchymal) mucin Blue
Nuclei Pink-red
Cytoplasm Pale pink

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. A brief dip in Solution A: Acetic Acid 3%, Aqueous from Step #3 can be added before Step #5 to remove excess Alcian Blue Stain.
    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. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 145-148.
      2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-175.
      3. Modifications developed by Newcomer Supply Laboratory.

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining umbilical cord.
Fixation: Formalin 10%, Phosphate Buffered (Part 1090).
Section/Glass: Paraffin sections cut at 4 microns on Superfrost™ Plus slides.
Quality Control Stain:  Alcian Blue pH 2.5 quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

With Alcian Blue 1%, pH 2.5 Stain Kit: Part 9102A/B Individual Stain Solution
Solution A: Acetic Acid 3%, Aqueous 250/500 ml Part 10017
Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous 250/500 ml Part 1003
Solution C: Nuclear Fast Red Stain, Kernechtrot 250/500 ml Part 1255

                                                                                                                             

APPLICATION:

Newcomer Supply Alcian Blue pH 2.5, Umbilical Cord Control Slides are for the positive histochemical staining of acid epithelial mucins (sialomucin, sulfomucin) as well as stromal (mesenchymal) mucin.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Heat dry sections in oven according to your laboratory protocol.
    2. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    3. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
    4. Move slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5 Aqueous. Stain for 30 minutes at room temperature or for 15 minutes in a 37°C water bath.
        1. See Procedure Note #3.
    5. Wash in running tap water for 10 minutes; rinse in distilled water.
    6. Counterstain in Solution C: Nuclear Fast Red Stain, Kernechtrot for 5 minutes.
        1. Shake solution well before use; do not filter.
    7. Rinse well in distilled water.
        1. See Procedure Note #4
    8. Dehydrate quickly through two changes of 95% ethyl alcohol and two changes of 100% ethyl alcohol. Clear in three xylene changes, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid epithelial mucins Blue
Stromal (mesenchymal) mucin Blue
Nuclei Pink-red
Cytoplasm Pale pink

 

PROCEDURE NOTES:

    1. Drain slides after each step to prevent solution carry over.
    2. Do not allow sections to dry out at any point during procedure.
    3. A brief dip in Solution A: Acetic Acid 3%, Aqueous from Step #3 can be added before Step #5 to remove excess Alcian Blue Stain.
    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. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 145-148.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 172-175.
    3. 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:  AFB, Ziehl-Neelsen quality control stained slide(s) included.
Reactivity: Guaranteed product specific reactivity for one year from date of receipt. Revalidate after one year to verify continued reactivity. 
Storage: 15-30°C in a light deprived and humidity controlled environment.
Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

With AFB, Ziehl-Neelsen Stain Kit: Part 9101A Individual Stain Solution
Solution A: Carbol Fuchsin Stain, Ziehl-Neelsen 250 ml Part 1030
Solution B: Acid Alcohol 1% 250 ml Part 10011
Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous 250 ml Part 12203

 

APPLICATION:

Newcomer Supply Acid Fast Bacteria (AFB), Animal Control Slides are for the positive histochemical staining of acid-fast mycobacteria in tissue sections.

 

PRESTAINING PREPARATION:

    1. Heat dry sections in oven according to laboratory protocol.
    2. Filter Solution A: Carbol Fuchsin Stain, Ziehl-Neelsen with filter paper when a thick sheen develops on solution surface.

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

    1. Deparaffinize sections thoroughly in three changes of xylene, 3 minutes each. Hydrate through two changes each of 100% and 95% ethyl alcohols, 10 dips each.  Wash well with distilled water.
        1. See Procedure Notes #1 and #2.
    2. Stain in Solution A: Carbol Fuchsin Stain, Ziehl-Neelsen for 30 minutes at room temperature. Keep solution covered.
        1. See Procedure Note #3.
    3. Rinse in running tap water for 2 to 3 minutes.
    4. Differentiate in Solution B: Acid Alcohol 1% until color no longer runs off slide and sections are pale pink; 3 to 10 rapid dips.
    5. Wash in running tap water 3 to 5 minutes; rinse in distilled water.
    6. Counterstain in Solution C: Light Green SF Yellowish Stain 0.1%, Aqueous; 2-5 dips.
    7. Rinse with one quick dip in distilled water or proceed directly to Step #10 without a distilled water rinse.
    8. Dehydrate in two changes each of 95% and 100% ethyl alcohol. Clear in three changes of xylene, 10 dips each; coverslip with compatible mounting medium.

 

RESULTS:

Acid-fast bacilli Bright red
Background Green

 

PROCEDURE NOTES:

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

 

REFERENCES:

    1. Carson, Freida L. and Christa Cappellano. Histotechnology: A Self-instructional Text. 5th ed. Chicago: ASCP Press, 2020. 213-215.
    2. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 237.
    3. Modifications developed by Newcomer Supply Laboratory.

 

Easily store your Slides and Tissue Paraffin Blocks in this sturdy and convenient system!

Stackable, corrugated cardboard file boxes that are an excellent value at a low cost!  Sturdy enough for transport and storage of blocks & slides.  The storage box has a double internal corrugated construction that supports up to 250+ lbs. and the trays remain easily accessible.

 

TISSUE PARAFFIN BLOCK STORAGE SYSTEM:

  • 8 drawers holds 500 blocks in a box
  • 5,000 block storage per case

 

SLIDE STORAGE SYSTEM:

  • 4 drawers hold 2,000 slides in a box
  • 20,000 slide storage per case

 

DIMENSIONS FOR BOTH SLIDE & BLOCK STORAGE BOXES:

  • 12″ (W) x 3 ¾”(H) x 11 ½” (L)

 

TISSUE PARAFFIN BLOCK & SLIDE STORAGE SYSTEM:

  • Well organized & detailed recording area on front panel of each box
  • Easily identify the slides or blocks contained in the box
  • From:/To: stickers available to place over recording area on front panel as well as on each drawer
  • Out/In place markers available to keep track of slides/blocks that have been removed
  • Easily stackable
  • Great pricing!
  • Made in the USA

 

Samples of accessories in each case!

 

This sturdy, heavy-duty, high capacity metal slide storage cabinet is an ideal solution for the safe, long-term storage of slide specimens. Each cabinet unit holds up to 4,500 slides. Up to 10 units can be secured together on a single base for a total of 45,000 slides per stack!

 

BUILD YOUR SLIDE FILING SYSTEM!

 

1.  The Base

  • Sturdy base can hold up to 10 cabinets
  • Available in Tan or Green

 

2. Slide Cabinet

  • 6 Drawers with brass pull knobs per cabinet.
  • Suspension design to help prevent accidental drawer pull out.
  • Cabinets lock together to protect against movement of cabinet and conserve space.
  • Each drawer section accommodates up to 4,500 slides.
  • Dimensions: 18 ¾” x 15 ¾” x 5″H
  • Available in Tan or Green.

 

3.  Stack Cabinets together to keep storage in one area.

 

BUILD YOUR SLIDE FILING SYSTEM!

 

1.  The Base

  • Sturdy base to hold 6 drawer slide cabinet.
  • Available in brown.

 

2. Slide Cabinet

  • 6 Drawers with black pull knobs per cabinet.
  • Cabinets lock together to protect against movement of cabinet and conserve space.
  • Each cabinet (6 drawers) accommodates up to 5,000 slides.
  • Each drawer comes with a foam stop block to keep slides standing upright when the drawer is not completely full.
  • Dimensions: 18 ¾” x 15 ¾” x 5″H
  • Available in brown.

 

3.  Stack Cabinets on base and together (up to 10 high) to keep storage in one area.

(use: Fite Stain for Nocardia & Leprosy.)

(use: Alcian Blue, pH 2.5.)

SOLUTION:

Gallon
Saccomanno Collection Fluid Part 1361B
60 ml vial, 30 ml fill (50/cs) 
Saccomanno Collection Fluid Vial Part 1361D

 

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

 

APPLICATION:

Newcomer Supply Saccomanno Collection Fluid is a ready-to-use cytology pre-fixative and sample preservative, and fluid of choice for collection of sputum specimens.  Saccomanno Collection Fluid also provides a variety of applications for collection and transport of cytology specimens, such as:

      • Fine needle aspirates (FNA)
      • Urine samples
      • Urinary tract washings
      • Bronchial washings and brushings
      • Bronchoalveolar lavage (BAL)
      • Gastrointestinal specimens

Saccomanno Collection Fluid works to partially fix and stabilize cytology specimens, when specimen transport is required or when cytopreparation is delayed.  Mucoproteins will remain suspended in the collection fluid and cells can be reclaimed with centrifugation.

 

METHOD:       

Fixation: Add equal volume of Saccomanno Collection Fluid to cytology specimen.

      • For brushes, add enough collection fluid to fully cover.

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

 

FIXATION PROCEDURE:

    1. After cytology specimen is collected and received in the laboratory, add equal volume of Saccomanno Collection Fluid to specimen that will be transported or held for an extended amount of time prior to cytopreparation.
        1. See Procedure Note #1.
    2. Store and/or transport specimen according to suggested temperature specifications for specimen type.
    3. Proceed with cytology sample fixation and processing method for specific specimen type.
        1. See Procedure Note #2.

 

PROCEDURE NOTES:

    1. Refer to laboratory guidelines for length of time a specific cytology specimen type can be held fresh prior to processing or addition of Saccomanno Collection Fluid.
    2. Saccomanno Collection Fluid is considered a cytology pre-fixative and not a substitute for final specimen fixation in appropriate strength alcohol.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-Instructional Text. 4th ed. Chicago: ASCP Press, 2015. 318-321.
    2. Kini, Sudha R. Color Atlas of Pulmonary Cytopathology. New York: Springer, 2002. 7-21.
    3. Koss, Leopold G. and Myron Melamed, eds. Koss’ Diagnostic Cytology and Its Histopathologic Bases. Fifth ed. Philadelphia: Lippincott Williams & Wilkins, 2006. 1570-1594.
    4. Modifications developed by Newcomer Supply Laboratory.

SOLUTION:

1 Liter 1 Gallon
Zenker Fixative, Modified, Zinc Chloride Part 1461A Part 1461B

 

Additionally Needed:

Acetic Acid, Glacial, ACS Part 10010

 

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

 

APPLICATION:

Newcomer Supply Zenker Fixative, Modified, Zinc Chloride is a mercury free alternative to the original Zenker formulation. Zenker Fixative, Modified, Zinc Chloride is a specialty fixative that provides crisp nuclear detail, frequently used for bone marrow clots and biopsies.

The acidic nature of Zenker Fixative, Modified, Zinc Chloride Working Solution may be enough to sufficiently decalcify bone marrow cores without additional decalcification steps.

 

METHOD:

Fixation: Recommended fixation is a minimum of 2 to 4 hours, depending upon tissue size and type.
Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

 

FIXATION PROCEDURE:

    1. Zenker Fixative, Modified, Zinc Chloride Working Solution:
        1. Zenker Fixative, Modified, Zinc Chloride      38 ml
        2. Acetic Acid, Glacial, ACS                 2 ml
    2. Combine and mix solutions directly before use. Fix tissue in fresh Zenker Fixative, Modified, Zinc Chloride Working Solution a minimum of 2 to 4 hours.
    3. Hold tissue specimens in Zenker Fixative, Modified, Zinc Chloride Working Solution for processing or a maximum of 24 hours.
        1. See Procedure Note #1.
    4. Wash fixed tissue in running tap water for minimum of 10 minutes to remove residual zinc chloride.
    5. Place on tissue processor in Formalin 10%, Phosphate Buffered (Part 1090) fixation step.
    6. Post-fixation applications of Zenker Fixative, Modified, Zinc Chloride include use as a mordant in phosphotungstic acid hematoxylin (PTAH) staining procedures.
        1. Refer to PTAH stain protocol for more information.

 

PROCEDURE NOTES:

    1. Extended storage in Zenker Fixative, Modified, Zinc Chloride is not recommended.
        1. After maximum fixation, rinse tissue in running tap water for a minimum of 10 minutes.
        2. Transfer Zenker fixed tissue to 70% Ethyl Alcohol (Part 10844) or Formalin 10%, Phosphate Buffered for long-term storage purposes.
    2. Zenker Fixative, Modified, Zinc Chloride is not recommended for preservation of red blood cells.
    3. Zenker Fixative, Modified, Zinc Chloride is corrosive, do not discard down the drain.
    4. Neutralize Zenker Fixative, Modified, Zinc Chloride with sodium carbonate or sodium bicarbonate to precipitate zinc at pH 7.0-8.0.
        1. Approximately 100 grams of sodium bicarbonate will neutralize/precipitate zinc from 1 liter of Zenker Fixative, Modified, Zinc Chloride.

 

REFERENCES:

    1. Carson, Freida L. and Christa Hladik. Histotechnology: A Self-Instructional Text. 3rd ed. Chicago, Ill.: American Society of Clinical Pathologists, 2009. 16-17, 20-21, 207-208.
    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. 148, 279.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 49.
    4. Modifications developed by Newcomer Supply Laboratory.

SOLUTION:

1 Liter 6 X 1 Liter 1 Gallon
B-5 Fixative Modified, Zinc Chloride Part 1015A Part 1015A Part 1015C

 

Additionally Needed:

Formaldehyde 37-40%, ACS Part 1089

 

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

 

APPLICATION:

Newcomer Supply B-5 Fixative Modified, Zinc Chloride is a mercury free alternative to the classic B-5 fixative.  It is the fixative of choice for bone marrow, lymph nodes, spleen and other hematopoietic tissues, providing clear nuclear detail while preserving immunohistochemical (IHC) staining.

 

METHOD:

Fixation Recommendations:

    • Bone Marrow Clots: Minimum of 2 hours.
    • Bone Marrow Biopsy: Minimum of 3 hours.
    • Lymph Nodes and Small Biopsies: Minimum of 4 hours.
    • Small nodes (5 mm or less) should be halved.
    • Larger nodes, dissected with no piece thicker than 3 mm.

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

 

FIXATION PROCEDURE:

    1. B-5 Fixative Modified, Zinc Chloride Working Solution:
        1. B-5 Fixative Modified, Zinc Chloride 50 ml
        2. Formaldehyde 37-40%, ACS                   5 ml
    2. Combine solutions directly before use. Fix tissue in fresh working solution a minimum of 2 to 4 hours.
    3. Hold tissue in B-5 Fixative Modified, Zinc Chloride Working Solution for processing or maximum of 72 hours.
        1. See Procedure Note #1.
    4. Wash fixed tissue in tap water a minimum of 10 minutes to remove residual zinc chloride.
    5. Decalcify bone marrow specimen as needed.
        1. See Procedure Note #2.
    6. Place on tissue processor in Formalin 10%, Phosphate Buffered (Part 1090) fixation step.

 

 PROCEDURE NOTES:

    1. After maximum fixation, transfer B-5 fixed tissue to 70% Ethyl Alcohol (Part 10844) for long-term storage.
    2. Nitric acid is not recommended as a decalcification agent following fixation in B-5 Fixative Modified, Zinc Chloride.
    3. Neutralize B-5 Fixative Modified, Zinc Chloride with sodium carbonate or sodium bicarbonate to precipitate zinc at pH 7.0-8.0.
        1. Approximately 100 grams of sodium bicarbonate will neutralize/precipitate zinc from 1 liter of B-5 Fixative Modified, Zinc Chloride.

 

REFERENCES:

    1. Bancroft, John D. and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 69.
    2. Carson, Freida L. and Christa Hladik Cappellano. Histotechnology: A Self-Instructional Text. 4th ed. Chicago: ASCP Press, 2015. 19-20.
    3. Dapson, Janet Crookham and Richard Dapson. Hazardous Materials in the Histopathology Laboratory: Regulations, Risks, Handling, and Disposal. 4th ed. Battle Creek, MI: Anatech, 2005. 148-149, 279.
    4. Modifications developed by Newcomer Supply Laboratory.

SOLUTION:

500 ml 1 Liter 1 Gallon
Wright Stain, Buffered Part 1422A Part 1422B Part 1422C

 

Additionally Needed:

Alcohol, Methanol Anhydrous, ACS Part 12236
Wright Stain Buffer, pH 6.8 Part 1430

 

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

 

APPLICATION:

Newcomer Supply Wright Stain, Buffered for Smears provides a quick staining technique for differential staining of cell types in peripheral blood smears as well as bone marrow smears/films.

 

METHOD:

Technique: Coplin jar or flat staining rack method
Solutions: All solutions are manufactured by Newcomer Supply, Inc.

 

PRESTAINING PREPARATION:

    1. Prepare a well-made blood smear or bone marrow smear/film with a focus on uniform cell distribution.
    2. Allow slides to thoroughly air-dry prior to staining.
    3. Filter Wright Stain, Buffered prior to use with quality filter paper.
        1. For flat staining rack method, filter sufficient stain to allow 1 ml of stain per slide.
    4. Prepare 25% Aqueous Methanol Rinse; combine and mix well.
        1. Distilled Water 30 ml  or  3 ml
        2. Methanol (Part 12236) 10 ml  or  1 ml

 

STAINING PROCEDURE:

    1. Coplin Jar Method: See Procedure Notes #1 and #2.
        1. Fix smears in Methanol for 15 seconds.
        2. Stain in filtered Wright Stain, Buffered for 1-2 minutes.
        3. Place directly into Wright Stain Buffer, pH 6.8 (Part 1430), for 1-4 minutes. Do Not Agitate!
        4. Dip quickly in 25% Aqueous Methanol Rinse (Step #4).
        5. Rinse in distilled water.
        6. Air-dry slides vertically; examine microscopically.
        7. If coverslip is preferred, air-dry slides and coverslip with compatible mounting medium.
    2. Flat Staining Rack Method: See Procedure Notes #1 and #2.
        1. Place slides on flat staining rack suspended over sink.
        2. Fix by flooding slide with Methanol for 15 seconds.
        3. Drain off Methanol.
        4. Flood each slide with 1 ml of filtered Wright Stain, Buffered for 1 minute.
        5. Retain Wright Stain, Buffered on slides.
        6. Directly add 2 ml of Wright Stain Buffer, pH 6.8 to each slide; agitate gently to mix with Wright Stain.
        7. Stain for an additional 3 minutes.
        8. Flood smears with 25% Aqueous Methanol Rinse (Step #4) for 1 second.
        9. Rinse in distilled water.
        10. Air-dry slides vertically; examine microscopically.
        11. If coverslip is preferred, air-dry slides and coverslip with compatible mounting medium.

 

RESULTS:

Erythrocytes Pink
Neutrophils Granules – Purple
Eosinophils Granules – Pink
White blood cells Chromatin – Purple
Lymphocytes Cytoplasm – Blue
Monocytes Cytoplasm – Blue
Bacteria Deep Blue

 

PROCEDURE NOTES:

    1. Timings are suggested ranges. Optimal staining times will depend upon staining intensity preference.
    2. Smears containing primarily normal cell populations require minimum staining time; immature cells and bone marrow smears/films may require longer staining time.
    3. The color range of stained cells may vary depending on buffer pH and pH of rinse water.
        1. Alkalinity is indicated by red blood cells being blue-grey and white blood cells only blue.
        2. Acidity is indicated by red blood cells being bright red or pink and lack of proper staining in white blood cells.
        3. If necessary, adjust buffer pH accordingly to 6.8 +/- 0.2.

 

REFERENCES:

    1. Lillie, R. D. and Harold Fullmer. Histopathologic Technic and Practical Histochemistry. 4th ed. New York: McGraw-Hill, 1976. 747-748.
    2. McPherson, Richard and Matthew Pincus. Henry’s Clinical Diagnosis and Management by Laboratory Methods. 22nd ed. Philadelphia: Elsevier Saunders, 2011. 522-532.
    3. Sheehan, Dezna C. and Barbara B. Hrapchak. Theory and Practice of Histotechnology. 2nd ed. St. Louis: Mosby, 1980. 154-155.
    4. Modifications developed by Newcomer Supply Laboratory.

 

Armadillo spleen with mycobacterium leprae the causative organism for leprosy.

 

Colon carcinoma 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).