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Electron Microscopy Sciences

Technical Data Sheets

Alcain Blue (pH 2.5) Method

EMS Catalog #: 26323-Series    


10% Buffered Neutral Formalin (#15740)


Paraffin at 6µm sections.

Staining Procedures:

  1. Deparaffinize and hydrate to distilled water.
  2. Mordant in Acetic Acid, 3% (#26323-03) for three minutes.
  3. Stain in Alcian Blue Solution (#26323-01) for thirty minutes. Wash in running water for ten minutes and rinse in distilled water.
  4. Counterstain in Nuclear Fast Red Solution (#16323-02) for five minutes. Wash in running water for one minute.
  5. Dehydrate in 95% Alcohol, Absolute Alcohol, and clear in Xylene (#23400), two changes each.
  6. Mount with Permount (#17986-01) or DPX (#13510).

Stain Results:

Weakly Acidic Sulfate Mucosubstances Dark Blue
Hyaluronic Acid Dark Blue
Sialomucins Dark Blue


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

Product Information

Alcain Blue (pH 2.5) Method