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10 October 2003

Microscopy: Hematoxylin & Periodic Acid-Schiff's Stain

Source: North Dakota State University

This is a good general staining procedure. It gives a nice pinkish cytoplasmic coloring and also stains basement membranes very strongly and thus give a lot of definition to the tissue. In addition, we are interested in microvessels, which have a well-defined basement membrane.

Recipies:

Schiff Reagent Solution

Composition:
1. Basic fuchsin-------------------------------------1gm (8gm)
2. Boiling water--------------------------------------100cc (800cc)
3. Potassium bisulfite-meta---------------------2gm (16gm)
4. 1N HCl-----------------------------------------------20cc (160cc)
5. Activated charcoal (powder)-----------------300mg (240mg)

Method in Lillie - Histopathologic Tech. And Pract-Histochem. 1954.

Bring water to a boil then remove from flame. At once add basic fuchsin, cool to 60°C. Filter, add potassium meta-bisulfite. Add 1N HCl, stopper and store solution in dark room at room temperature for 18-24 hours. Add charcoal (activated), shake for 1 minute. Filter and store at 0-5°C. Solution should be clear with no color to a light yellow color. Discard when it becomes pink.