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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
- De-paraffin slides in xylene (1) ———— 2 minutes (re-use)
- De-paraffin slides in xylene (2)———— 2 minutes (re-use)
- Clear slides in 100% alcohol————— 2 minutes
- Clear slides in 100% alcohol————— 2 minutes
- Hydrate slides in 95% alcohol————- 2 minutes
- Hydrate slides in running water———- 5 minutes
- Place slides in 3% Acetic Acid————- 3 minutes (re-use)
- Alcian Blue ———————————– 30 minutes (re-use)
- Wash in running water ——————— 10 minutes
- Oxidize in 0.5% periodic acid ————- 10 minutes (re-use)
- Wash in running water ——————— 5 minutes
- Schiff reagent (room temp)—————- 20 minutes (re-use)
- Wash in running water ——————— 10 minutes
- Dehydrate in 95% alcohol—————— 30 seconds
- Dehydrate in 100% alcohol—————- 1 minute
- Dehydrate in 100% alcohol—————- 2 minutes
- Dehydrate in 100% alcohol—————- 2 minutes
- Clear in xylene (3)————————— 2 minutes (re-use)
- 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
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