by Histology Reference Guide

Alcian Blue / PAS Stain Protocol

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Here is the protocol for alcian blue periodic acid schiff  (AB /PAS).  At the bottom of the post is the link for printing.

Control tissue: Large Intestine

  1. De-paraffin slides in xylene (1) ———— 2 minutes (re-use)
  2. De-paraffin slides in xylene (2)———— 2 minutes (re-use)
  3. Clear slides in 100% alcohol————— 2 minutes
  4. Clear slides in 100% alcohol————— 2 minutes
  5. Hydrate slides in 95% alcohol————- 2 minutes
  6. Hydrate slides in running water———-  5 minutes
  7. Place slides in 3% Acetic Acid————- 3 minutes (re-use)
  8. Alcian Blue ———————————– 30 minutes (re-use)
  9. Wash in running water ——————— 10 minutes
  10. Oxidize in 0.5% periodic acid ————- 10 minutes (re-use)
  11. Wash in running water ——————— 5 minutes
  12. Schiff reagent (room temp)—————- 20 minutes (re-use)
  13. Wash in running water ——————— 10 minutes
  14. Dehydrate in 95% alcohol—————— 30 seconds
  15. Dehydrate in 100% alcohol—————- 1 minute
  16. Dehydrate in 100% alcohol—————- 2 minutes
  17. Dehydrate in 100% alcohol—————- 2 minutes
  18. Clear in xylene (3)————————— 2 minutes (re-use)
  19. Clear in xylene (4)————————— 5 minutes (re-use)

Results: Polysaccharides and Mucosubstances containing hexoses or deoxyhexoses with vicinal glycol groups – Magenta to Red

Hyaluronic acid, sialomucins and all but the most strongly acidic sulfated mucosubstances – Blue

PAS Alcian Blue

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