Bosma's Alum Hematoxylin
Materials
Material | Amount | Function |
---|---|---|
Hematoxylin, 10% alc. | 25 mL | Dye |
Ammonium alum | 25 g | Mordant |
Tap water | 865 mL | Solvent |
Diethylene glycol | 100 mL | Stabiliser |
Sodium iodate | 0.2 g | Oxidant |
Glacial acetic acid | 10 mL | Acidifier |
Compounding procedure
- Place 815 mL hot tap water in a large flask.
- Add the ammonium alum, mix well to dissolve and cool to room temperature.
- Add the alcoholic hematoxylin solution (ripened).
- Add the sodium iodate dissolved in 50 mL cold tap water and mix well.
- Add the diethylene glycol and mix well.
- Add the acetic acid and mix well. The pH should be 3.1 to 3.3 and the solution may be used immediately.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for an appropriate time.
- Rinse well with water.
- Differentiate with acid ethanol if necessary.
- Rinse with water and blue.
- Rinse well with water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Background – as counterstain or unstained
Notes
- The staining time should be determined by trial.
- Acid ethanol is 0.5% – 1% hydrochloric acid in 70% ethanol.
- Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Bosma, Rob
A useful hematoxylin without toxic chemicals.
Histologic, V 18, Nº 1, January 1988