Galigher's Alum Hematoxylin
This solution was recommended as a substitute for Harris’ Alum hematoxylin.
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 5 g | Dye |
| Ammonium alum | 3 g | Mordant |
| Distilled water | 500 mL | Solvent |
| 100% ethanol | 500 mL | Solvent |
| Mercuric oxide | 6 g | Oxidant |
Compounding procedures
- Combine the water and ethanol.
- Add the Alum and dye, and bring to a boil.
- Add the mercuric oxide, then simmer for 20 minutes.
- Restore the volume to 1 L with 50% ethanol.
- Cool, and filter through double thickness filter paper.
- Store in a tightly stopper bottle.
- The solution may be used immediately, and is stable for about six months.
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
- Mercuric salts are toxic and no longer recommended for oxidation of hematoxylin. Ensure that discarded solution is disposed of as mercury contaminated, or substitute 0.5 grams sodium iodate as the oxidant.
- Staining capability may continue to improve for a short while.
- The appropriate 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
- Bolles Lee, A.. Edited by Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist’s Vade-Mecum. 11 ed.,
Churchill, London, UK.
