Alizarin Red for Calcium Deposits

EMS26206

Components:

Alizarin S 2% Solution pH 4.2

Reagent Alcohol 50%

Acetone

Acetone-Xylene

Fixation:

Alcoholic Formalin

Sections:

Paraffin, Ester Wax, Celloidin or Gelatin

Staining Procedures:

1. Bring sections to Alcohol, 50% and rinse rapidly in distilled water.
2. Cover sections with Alizarin Red S and follow the staining by transmitted light, on the stage of a staining microscope. Calcium sites will be covered with a vivid orange-red calcium-alizarin lake within a period of 30 seconds to 5 minutes. The end point should be chosen when the deposit is heavy but not too diffused.
3. Shake off excess stain and blot carefully with filter paper or alternatively drain very thoroughly.
4. Place immediately into Acetone for 10 – 20 seconds followed by Acetone-Xylene for 10 -20 seconds.
5. Clear in Xylene and mount in Depex or Cedar wood Oil 

Stain Results:

References:

McGee-Russell, S.M., J. Histochem., 6:22-42, 1958.

Clark, G., (ed.). Staining Procedures, 3rd ed., Williams & Wilkins, Baltimore, p. 140, c. 1973.