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PAS (Periodic Acid Schiff) Staining Protocol
Description: This method is used for detection of glycogen in tissues such as liver, cardiac and skeletal muscle on formalin-fixed, paraffin-embedded tissue sections, and may be used for frozen sections as well. The glycogen, mucin, and fungi will be stained purple and the nuclei will be stained blue.
Fixation:
10%
formalin.
Section:
paraffin sections at 5 um.
Solutions and Reagents:
0.5% Periodic
Acid Solution:
Periodic acid ---------------------- 0.5 g
Distilled water -------------------- 100 ml
Schiff Reagent:
Test for Schiff
reagent: Pour 10 ml of 37% formalin into a watch glass. To this add
a few drops
of the Schiff reagent to be tested. A good Schiff reagent will
rapidly turn a red-purple color. A deteriorating schiff reagent will
give a delayed reaction and the color produced will be a deep
blue-purple.
Mayer’s
Hematoxylin Solution:
Procedure:
1. Deparaffinize and hydrate to water.
2. Oxidize in 0.5% periodic acid solution for 5 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
Mayer's hematoxylin for 1
minute.
7. Wash in tap water for 5 minutes.
8. Dehydrate and coverslip using a synthetic mounting medium.
Results:
Glycogen, mucin and some basement membranes --- red/purple
Fungi
------------------------------------------------------
red/purple
Background ----------------------------------------------- blue