Kefalas's Iron Hematoxylin
Materials
Solution A
Material | Amount | Function |
---|---|---|
Hematoxylin | 1 g | Dye |
Ferric chloride | 1 g | Mordant |
Acetone | 100 mL | Solvent |
Hydrochloric acid | 0.05 mL | Acidifier |
Compounding Procedure
- Dissolve the ferric chloride and hematoxylin in the acetone.
- Add the hydrochloric acid.
Protocol
Standard Method
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 30 minutes.
- Wash well in running tap water to blue.
- Rinse with distilled water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Alternative Method
Since this solution is almost anhydrous it can be used in a staining procedure which completely avoids water or ethanol.
- Remove wax with xylene.
- Remove xylene with a few changes of acetone.
- Place into staining solution for 30 minutes.
- Wash with acetone.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Background – as counterstain or unstained
Notes
- 0.05 mL hydrochloric acid would be about 1-2 drops.
- The simplicity of the formula indicates it is likely not stable for long.
- The presence of hydrochloric acid may indicate progressive staining.
- It is likely suitable as an acid resistant nuclear stain.
- Although the staining time should be established by trial and error, 30 minutes would likely suffice as a starting point.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
Citing:
Kefalas, (1926),
Journal of the Royal Microscopical Society
v. 46, p. 277.