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Csaba’s Stain for Mast Cells

Csaba's Stain

for Mast Cells

5
steps
6
materials

Materials

Staining solution

MaterialAmount
Alcian blue0.36g
Safranin0.18g
Ferric ammonium sulfate0.48g
Walpole’s acetate HCl buffer100mL

Walpole’s acetate-HCl buffer, pH 1.42

MaterialAmount
Sodium acetate, M/1100mL
Hydrochloric acid, M/1120mL

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

  1. Bring sections to water via xylene and ethanol.
  2. Place in to staining solution for 10-20 minutes.
  3. Rinse with water.
  4. Dehydrate with tertiary butanol.
  5. 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

  1. Culling, C F A, (1974).
    Handbook of Histopathological and Histochemical Techniques., Ed. 3. pp. 169 & 419
    Butterworths, London, England.