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.

Stain Results:

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

Magenta to red

Mucosubstances include neutral mucosubstances

Red

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

Blue

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

Magenta to red

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.