Csaba's Stain
for Mast Cells
Materials
Staining solution
Material | Amount | |
---|---|---|
Alcian blue | 0.36 | g |
Safranin | 0.18 | g |
Ferric ammonium sulfate | 0.48 | g |
Walpole’s acetate HCl buffer | 100 | mL |
Walpole’s acetate-HCl buffer, pH 1.42
Material | Amount | |
---|---|---|
Sodium acetate, M/1 | 100 | mL |
Hydrochloric acid, M/1 | 120 | mL |
Tissue Sample
Paraffin sections of neutral buffered formalin fixed tissue are suitable. Mercuric chloride fixatives are reputed to emphasise metachromasia. Other fixatives may be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place in to staining solution for 10-20 minutes.
- Rinse with water.
- Dehydrate with tertiary butanol.
- clear with xylene and mount using a resinous medium.
Expected Results
- Young mast cells – blue
- Mature mast cells – red
Notes
- To make an M/1 solution of sodium acetate, dissolve 82.04 grams anhydrous or 136.09 crystal sodium acetate in one litre of distilled water.
- To make an M/1 solution of hydrochloric acid, dilute concentrated hydrochloric acid, which is about 10M, with distilled water (one part acid to 8 parts water) and titrate against an alkali of known molarity. Adjust the concentration to M/1 with distilled water.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling, C F A, (1974).
Handbook of Histopathological and Histochemical Techniques., Ed. 3. pp. 169 & 419
Butterworths, London, England.