Anderson's Iron Hematoxylin
Materials
Solution A
Material | Amount | Function |
---|---|---|
Hematoxylin | 0.5 g | Dye |
100% ethanol | 50 mL | Solvent |
Distilled water | 50 mL | Solvent |
Calcium hypochlorite, 2% aqueous | 5 mL | Oxidant |
Solution B
Material | Amount | Function |
---|---|---|
Ferric ammonium sulphate | 3 g | Mordant |
Distilled water | 100 mL | Solvent |
Sulfuric acid | 0.5 mL | Acidifier |
Compounding procedures
- Make each solution separately.
- For use, add 2 volumes of solution A to 1 volume of solution B.
- The working solution may be used immediately, but is not stable for long.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for an appropriate time.
- 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.
Expected Results
- Nuclei – black
- Background – as counterstain or unstained
Notes
- The stock solutions are stable for some time.
- The working solution should be made fresh.
- It is claimed that the working solution rarely overstains, i.e. it is progressive.
- Solution A incorporates calcium hypochlorite as an oxidizing agent for hematoxylin. Presumably other oxidizing agents would suffice equally as well. Sodium iodate (0.1 g or less) is the most common.
- The staining time should be determined by trial.
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:
Anderson, J., (1929)
How to stain the nervous system.
Livingstone. Edinburgh, Scotland.