Protocol
Allen's Stain
for Mast Cells
Materials
- Mayer’s hemalum
- Neutral red
Material Amount Neutral red 0.5 g Ethanol, 50% 100 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.
- Rinse with water.
- Stain nuclei lightly with alum hematoxylin.
- Rinse with tap water and blue hematoxylin.
- Rinse well with water.
- Place in neutral red for 10 minutes.
- Rinse with distilled water.
- Differentiate with 70% ethanol up to 10 minutes.
- Dehydrate with 96% ethanol up to 5 minutes.
- Dehydrate with N-butanol up to 10 minutes.
- Clear with xylene and mount using a resinous medium.
Expected Results
- Nuclei – blue
- Mast cell granules = red
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling, C F A, (1976).
Lynch’s Medical Laboratory Technology., Ed. 3. Vol. II. pp. 980
W. B. Saunders Company, Toronto, Canada. - Putt, F A, (1972).
Manual of Histopathological Staining Methods., pp. 233
John Wiley & Sons, London, UK.