Periodic Acid Schiff (PAS) Glycogen Staining Protocol

 

Fixation: 10% formalin for 10 minutes.

 

Section: paraffin sections at 6 micro meter.

 

Solutions and Reagents:

 

0.5 % Periodic Acid Solution:

      Periodic acid  ------------------- 0.5 g

      Distilled water ------------------100 ml

Schiff Reagent

Hematoxylin Solution:

 

Procedure:

 

1.    Deparaffinize and rehydrate.

2.    Rinse in 0.5% periodic acid solution for 10 minutes.

3.    Rinse in distilled water.

4.    Place in Schiff reagent for 15 minutes (Sections become light pink color during this step).

5.    Wash in lukewarm tap water for 5 minutes (Immediately sections turn dark pink color).

6.    Counterstain in hematoxylin for 30 seconds.

7.    Wash in tap water for 5 minutes.

8.    Dehydrate and mount with a coverslip.

 

Results:

 

      Glycogen, mucin and some basement membranes --- red/purple

      Background ----------------------------------------------- blue

 

PAS reactions ¨C Reaction intensities will vary; however, the following substances will be PAS-positive (magenta): glycogen, neutral mucosubstances, basement membranes, collagen fibers, fungal cell walls, and glycoproteins such as yolk vesicles and phospholipids.