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.
1.Deparaffinize slides and rehydrate to distilled water.
2.Staining in alcian blue solution for 10-15 min, wash in running tap water for 2 min.
3.Dehydrate through absolute alcohol for 3 times, 5min each time,clear in xylene for 2 times, 5min each time,mount with neutral gum.
4.Microscopy detection 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.