Bullard's Alum Hematoxylin
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 8 g | Dye |
| Ammonium alum | 20 g | Mordant |
| Distilled water | 250 mL | Solvent |
| 50% ethanol | 144 mL | Solvent |
| Glacial acetic acid | 16 mL | Acidifier |
| Mercuric oxide, red | 8 g | Oxidant |
| 95% ethanol | 275 mL | Solvent |
| Glycerol | 330 mL | Stabiliser |
| Glacial acetic acid | 18 mL | Acidifier |
| Ammonium alum | 40 g | Mordant |
Compounding procedure
- Combine the first five ingredients and bring to a boil.
- Add the mercuric oxide with caution.
- Cool and filter.
- Add the last four ingredients.
- The solution may be used immediately, and is stable for more than a year.
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 solution is likely regressive
- The appropriate 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
- F. A. Putt
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA
