Alcian Blue/PAS Stain Kit

alcian blue PAS stain kit periodic acid schiff

Product Options:

Part # 91022A250ml kit $181.10
Part # 91022B500ml kit $325.20

 

  

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

 

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

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

Additionally Needed:

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

 

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

 

APPLICATION:

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

 

METHOD:

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

Technique:   Paraffin sections cut at 5 microns

Solutions:     All solutions 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.

 

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.

a. See Procedure Notes #1 and #2.

  1. Place slides in Solution A: Acetic Acid 3%, Aqueous for 3 minutes.
  2. Place slides directly into Solution B: Alcian Blue Stain 1%, pH 2.5  Aqueous for 30 minutes.
  3. Wash slides in gently running tap water for 10 minutes; rinse in distilled water.
  4. Place slides in Solution C: Periodic Acid 0.5%, Aqueous for 10 minutes.
  5. Wash slides in running tap water for 5 minutes; rinse in distilled water.
  6. Place slides in Solution D: Schiff Reagent, McManus for 20 minutes.
  7. Wash in lukewarm tap water for 5-10 minutes.
  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 epithelial mucin Violet
Neutral epithelial mucin Magenta
Glycogen Magenta
Stromal (mesenchymal) mucin Blue

 

PROCEDURE NOTES:

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

     

REFERENCES:

  1. Bancroft, John D., and Marilyn Gamble. Theory and Practice of Histological Techniques. 6th ed. Oxford: Churchill Livingstone Elsevier, 2008. 173-174.    
  2. Carson, Freida, Histotechnology: A Self-Instructional Text. 2nd ed. Chicago: ASCP Press, 1997. 91-102, 121-123.    
  3. Modifications developed by Newcomer Supply Laboratory.