Csaba's Stain
for Mast Cells

Solutions

Staining solution
Alcian blue 0.36 g
Safranin 0.18 g
Ferric ammonium sulphate 0.48 g
Walpole's acetate HCl buffer 100 mL
Walpole's acetate-HCl buffer, pH 1.42
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.

Method

  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 Mature
Mast cells Blue Red

Notes

  1. 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.
  2. 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.

Reference

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

 


 

 

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