Alcian Blue/PAS

Validation Stain: Alcian Blue/PAS

 

Tissue: 
(+) Small Intestine
Adhesive or Charged Slides: 
Superfrost +

Product Options:

Part # 4022A 10/Set $52.00
Part # 4022B 98/Set $365.00

PRODUCT SPECIFICATIONS:

Tissue:  Positive staining small intestine.

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

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

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

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

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

Intended Use: To verify histological techniques and reagent reactivity.

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

 

CONTROL SLIDE VALIDATION:

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

 

APPLICATION:

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

 

NEWCOMER SUPPLY VALIDATION PROCEDURE:

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

 

RESULTS:

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

 

PROCEDURE NOTES:

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

 

REFERENCES:

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