Description: 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. Strongly acidic mucosubstances will be stained blue, nuclei will be stained pink to red, and cytoplasm will be stained pale pink.
Fixation: formalin fixed, paraffin embedded tissue sections.
Solutions and Reagents:
3% Acetic Acid Solution:
Glacial acetic acid ----------------- 3 ml
Alcian Blue Solution (pH 2.5):
Acetic acid, 3% solution ----------- 100 ml
Mix well and adjust pH to 2.5 using acetic acid.0.1% Nuclear Fast Red Solution:
Nuclear fast red ------------------- 0.1 g
Dissolve aluminum sulfate in water. Add nuclear fast red and slowly heat to boil and cool. Filter and add a grain of thymol as a preservative.
Procedure:
1. Deparaffinize slides and hydrate to distilled water.
2. Stain in alcian blue solution for 30 minutes.
3. Wash in running tap water for 2 minutes.
4. Rinse in distilled water.
5. Counterstain in nuclear fast red solution for 5 minutes.
6. Wash in running tap water for 1 minute.
7. Dehydrate and through 95% alcohol, 2 changes of absolute alcohol, 3 minutes each.
8. Clear in xylene or xylene substitute.
9. Mount with resinous mounting medium.
Results:
Strongly acidic sulfated mucosubstances -------------- blue
Positive Controls:
Small intestine, appendix, or colon.
