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Alcian Blue/PAS Stain Kit

Histological Stain Kit Alcian Blue/PAS

Differentiates between acidic epithelial mucins (sialomucin, sulfomucin) and neutral epithelial mucin and is a means of detecting the overall presence of mucins.  Acidic mucins are stained with the Alcian Blue technique while neutral mucins and glycogen are stained by the PAS reaction.

Product Options:

Part # 91022A 250 ml kit $209.40
Part # 91022B 500 ml kit $360.80

ALCIAN BLUE/PAS STAIN KIT INCLUDES:

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

           

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

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

Additionally Needed:

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


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

 

APPLICATION:

Newcomer Supply Alcian Blue/PAS Stain Kit procedure is used to differentiate acidic epithelial mucins (sialomucin, sulfomucin), neutral epithelial mucins and stromal (mesenchymal) mucins and is a means of detecting the overall presence of mucins.

 

METHOD: 

Fixation: Formalin 10%, Phosphate Buffered (Part 1090)
Technique:  Paraffin sections cut at 4 microns
Solutions:  All solutions are manufactured by Newcomer Supply, Inc.

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

 

STAINING PROCEDURE:

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

 

RESULTS:

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

 

PROCEDURE NOTES:

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

 

REFERENCES:

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