Alcian Blue and PAS Stain

Principle:

Neutral mucin contains aminohexose and free hexose. Acid mucin also contains aminohexose and different kinds of acid radical. Neutral mucins are found in gastric mucosa, duodenum gland, submandibular gland and prostatic epithelium. Acid mucins are found in respiratory tract, goblet cells of digestive tract, aortic wall, cartilage matrix, cornea and skin. Gastric mucosa secretes neutral mucus. Acid mucins are secreted in intestinal metaplasia of stomach or intestinal type of gastric cancer. However, gastric type of gastric cancer secretes neutral mucus. Acid and neutral mucins are hardly distinguished in hematoxylin and eosin stain. With AB-PAS stain, acid mucin can be stained blue by Alcian stain. Neutral musin can be stained red by PAS.

 

Specifications:

 

Contents	4vials×20ml/Kit	Components
Alcian blue solution（pH2.5）	1vial×20ml	Alcian blue
Periodic Acid Solution	1vial×20ml	Periodic Acid
Schiff Reagent	1vial×20ml	Fuchsin
Mayer Hematoxylin solution	1vial×20ml	Hematoxylin

 

Methods:

1. Deparaffinize tissue in deionized water. Rinse briefly with distilled water.

2. Place in Alcian Blue solution (pH2.5) for 10~20 minutes.

3. Rinse briefly with water.

4. Oxidize tissue in Periodic acid for 10 minutes.

5. Rinse in distilled water for 2 minutes.

6. Place in Schiff reagent for10~15 minutes.

7. Wash in running tap water for 5 minutes.

8. Stain nuclei in Mayer Hematoxylin solution for 1~2 minutes.

9. Wash in running tap water.

10. Dehydrate, clear and mount by following routine procedure.

 

Expected Results:

Acid mucus……………….Blue

Neutral mucus…………….Red

Mixed mucus……………...Reddish-purple

Nucleus………...………….Light blue.

 

Precaution:

1. Store the kit in the refrigerator, and bring it to room temperature to equilibrate the reagents for 30 minutes before use.

2. Tightly cap Schiff reagent after use. When rosiness appears, discard the reagent.

3. Glycogen appears red. It is easy to differentiate glycogen and mucous substance from the morphology and distribution.

4. Light counterstain of nuclei should be used for differentiation with that of Alcian Blue stain. This procedure can also be omitted.