McGhee Russel
Alizarin Red S for Calcium

Solutions

Staining solution
Alizarin red S	2	g		Ajust the pH to 4.2 with 10% ammonium hydroxide.
Distilled water	100	mL		Use a pH meter with a glass electrode.

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory. Fixatives containg acids may remove some calcium.

Method

1. Bring sections to distilled water via xylene and ethanol.
2. Place into the staining solution for 1-5 minutes.
3. Blot.
4. Rinse well with acetone.
5. Rinse with acetone:xylene 1:1.
6. Rinse with xylene and mount with a resinous medium.

Expected results

• Calcium  –  orange to red

Notes

1. Although this method demonstrates calcium itself, the von Kossa method is often preferred.
2. Stained calcium is birefringent.
3. The staining should be controlled visually.

 

Reference

Bancroft, J.D. and Stevens A. (1982)
Theory and practice of histological techniques Ed. 2
Churchill Livingstone, Edinburgh & London, UK.