Friday, 10 June 2011
Alcian Blue Stain For Acidic Mucins – Method and Tips
The alcian blue stain is this author’s preferred stain for the demonstration of acidic mucins. The dye was originally used for the dyeing of cotton before being discovered as a by Steedman in 1950.
The alcian blue itself is a cationic copper phthalocyanine dye which stains mucopolysaccharides and glycosaminoglycans a bluish colour. Within skin, acidic mucins can be found in many differing conditions such as a mucinoma, lupus and alopecia mucinosa.
Below is the preferred alcian blue method of this author
1g alcian blue in 3% acetic acid (check pH = 2.5)
1. Take sections to water
2. Cover slide with FILTERED alcian blue solution and leave for 20 minutes
3. Rinse in tap water
4. Counterstain with FILTERED 1% safranin for 10-15 seconds
5. Rinse in tap water
6. Dehydrate quickly, clear and mount.
- this author prefers an alcian blue staining time of about 20 minutes but can be done within the range of 10-30 minutes if desired.
- by reducing the pH to 0.2 the stainer can select for only strongly sulphated mucins. A pH of 1.0 stains both weak and strongly sulphated mucins. If using a lower pH method be sure no to rinse in tap water between the steps for too long as this can affect the alcian blue staining.
- this author prefers safranin as a counterstain due to its crisper staining, but safranin leeches out quickly in the dehydrating alcohols therefore blot dry after counterstaining and quickly dehydrate through the alcohols.
- neutral red can also be used as a counterstain but this author prefers safranin as it stains quicker and has a greater contrasting colour.
- the alcian blue staining solution expires after approximately 6 months.
Thanks for reading and I welcome any comments and other tips.
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