PURPOSE:
Alcian blue stains acid mucosubstances and acetic mucins.
Excessive amounts of non-sulfated acidic mucosubstances are seen in
mesotheliomas, certain amounts occur normally in blood vessel walls but
increase in early lesions of atherosclerosis.
PRINCIPLE:
Alcian blue is a group of polyvalent basic dyes that are water
soluble. The blue color is due to the presence of copper in the molecule.
The 3% acetic acid solution (pH2.5), Alcian blue stains both sulfated and
carboxylated acid mucopolysaccharides and sulfated and carboxylated
sialomucins (glycoproteins). It is believed to form salt linkages with the
acid groups of acid mucopolysaccharides.
PROCEDURE
1. Deparaffinize sections, if required, and hydrate to
distilled water.
2. Apply Acetic Acid Solution onto tissue section for
3 minutes.
3. Tip off excess and apply Alcian Blue Solution for
30 minutes at room temperature or 15 minutes at
37oC.
4. Rinse slide briefly (10-30 seconds) in Acetic Acid
Solution to remove excess Alcian Blue.
5. Rinse for 2 minutes in running tap water followed by
2 changes of distilled water (1-2 minutes each).
6. Apply Nuclear Fast Red Solution for 5 minutes.
7. Rinse for 2 minutes in running tap water followed by
2 changes of distilled water (1-2 minutes each).
8. Dehydrate through graded ethanols.
9. Clear and coverslip.
Acidic Sulfated Mucosubstances: Blue
Hyaluronic Acid: Blue
Sialomucins: Blue
Nuclei: Red
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