PAS-Alcian Blue Method For Mucosubstances

Components:

Alcian Blue Solution

Periodic Acid 1%

Schiff's Reagent

Sodium Metabisulphite 0.5% aqueous

Fixation:

10% Buffered Neutral Formalin

Section:

Paraffin, @ 6µm

Staining Procedure:

1. Deparaffinise and hydrate to distilled water.
2. Stain in Alcian Blue Solution, pH 1.0 or Alcian Blue, pH 2.5 for 30 minutes.
3. If Alcian Blue solution, pH 1.0 is used, blot section dry with filter paper. If Alcian Blue solution pH 2.5 is used wash, in water for 5 minutes.
4. Oxidize in Periodic Acid 1% for 30 minutes. Wash in running water for 5 minutes.
5. Schiff's Reagent, for 10 minutes.
6. Rinse in Sodium Metabisulfite, 0.5% aqueous, three changes, two minutes each. Wash in running water for ten minutes.
7. Dehydrate in 95% Alcohol, Absolute Alcohol and clear in Xylene, two changes each.
8. Mount with Permount.

Results:

PAS-Alcian Blue pH 2.5 - All polysaccharides and mucosubstances containing hexoses or deoxyhexoses with vicimal glycol groups stain magenta to red.

Mucosubstances staining red include neutral mucosubstances.

Hyaluronic acid, sialomucins and all but the most strongly acidic sulphated mucosubstances stain blue.

PAS-Alcian Blue pH 1.0 - PAS positive same as above. Alcinophilic substances at this pH include only the sulphated mucocaccharides.

References:

AFIP Manual of Histological Staining Methods, 3 rd ed. Ed. L. Luna: New York :

McGraw Hill Publications c. 1968, p. 168.

Lev. R. & Spicer S.S. J. Histochem. Cytochem., 12:309. 1964.