Alcian blue is a cationic dye, stains acid mucosubstances and acetic mucins specifically. Acid mucosubstances in cytoplasm are stained blue by alcian blue. Alcian blue solution (pH=2.5) can be used for staining cartilage tissue, or combined with periodic acid-schiff staining.
Alcian Blue Stain
1 Deparaffinize slides and rehydrate to distilled water
2 Stain in alcian blue solution for 10-15 min, wash in running tap water for 2 min.
3 Dehydrate through 3 changes of absolute alcohol, 5 min each. Clear in 2 changes of xylene, 5 min each. Mount with resinous mounting medium.
4 Microscopy examination and analysis.
For cartilage tissue, general mucins which contain carbonyl and weak acid mucous matters are stained blue, the strong acidic mucosubstances are pale or non-stain. Nucleus and background are unpigmented.
If counterstain is necessary, Nuclear Fast Red solution can be used to stain nucleus. Combine with PAS Stain solution (G1008) is also alternative.
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