Bennett's Alum Hematoxylin
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 1 g | Dye |
| Potassium alum | 90 g | Mordant |
| Distilled water | 1 L | Solvent |
| Sodium iodate | 0.2 g | Oxidant |
| Chloral hydrate | 50 g | Stabiliser |
| Citric acid | 1 g | Acidifier |
Compounding procedure
- Heat the water. Then add in order:
- Hematoxylin
- Sodium iodate
- Alum
- Citric acid
- Chloral hydrate.
Note: Dissolve each ingredient before adding the next.
- Cool to room temperature and filter. It may be used immediately.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 5-20 minutes.
- 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
- Putt gives no information regarding staining time and characteristics, but the similarity of the formula to Mayer’s (Langeron’s) hemalum suggests it is progressive and selectively nuclear. A staining time of 10-20 minutes should be adequate.
- 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
- F. A. Putt
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA
