Alcain Blue (pH 2.5) Method

Created by Kathleen Patrick, Modified on Fri, 8 Nov at 1:40 PM by Kathleen Patrick

EMS26323


Components:

Alcian Blue 1%

Nuclear Fast Red Solution

Acetic Acid 3% Aqueous


Fixation:

10% Buffered Neutral Formalin


Sections:

Paraffin at 6µm sections.


Staining Procedures:

  1. Deparaffinise and hydrate to distilled water.
  2. Mordant in Acetic Acid, 3% for three minutes.
  3. Stain in Alcian Blue Solution for thirty minutes. Wash in running water for ten minutes and rinse in distilled water.
  4. Counterstain in Nuclear Fast Red Solution for five minutes. Wash in running water for one minute.
  5. Dehydrate in 95% Alcohol, Absolute Alcohol, and clear in Xylene, two changes each.
  6. Mount with Permount or DPX.


Stain Results:

Weakly Acidic Sulfate Mucosubstances

Dark Blue

Hyaluronic Acid

Dark Blue

Sialomucins

Dark Blue


References:

AFIP Manual of Histological Staining Methods, 3rd ed., Ed. L. Luna: New York: McGraw Hill Publications, c. 1968, p. 163.

Lev, R. & Spicer, S.S., J. Histochem. Cytochem., 12:309, 1964

 

 

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