ALIZARIN RED S FOR CALCIUM IN PARAFFIN SECTIONS
· May be used alone or in conjunction with the Alcian Blue stain for acid mucosubstances.
PROTOCOL:
1. De-paraffinize and hydrate slides to distilled water.
2. Alizarin Red S Solution, 30 sec – 5 min. Observe reaction under a microscope and remove staining solution as soon as a red-orange lake is formed.
3. Shake off excess dye and blot carefully.
4. Dehydrate in acetone, 10 – 20 sec.
5. Acetone/Xylenes, 10 – 20 sec.
6. Clear in xylenes & coverslip using Cytoseal XYL.
Results:
Calcium deposits (except oxylate)– red-orange (birefringent)
SOLUTIONS:
1. Alizarin Red S Solution:
Alizarine Red S (C.I. 58005) 2 gm
Distilled water 100 ml
Mix & adjust to pH 4.2 with dilute ammonium hydroxide.
May be made in advance.
2. Acetone/Xylenes:
Acetone 50 ml
Xylenes 50 ml
May be made in advance.