Pusey's Alum Hematoxylin
This solution is described as a modification of Mayer’s hemalum, but please refer to that formula.
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin, aged 10% ethanolic | 16 mL | Dye |
| Ammonium alum | 60 g | Mordant |
| Distilled water | 1 L | Solvent |
| Sodium iodate | 0.25 g | Oxidant |
| Citric acid, 5% aqueous | 7 mL | Acidifier |
| Chloral hydrate | 50 g | Stabiliser |
Compounding Procedure
- Dissolve the Alum in the water using low heat, but do not boil.
- Add the hematoxylin solution and mix well.
- Add the sodium iodate and dissolve.
- Leave for 30 minutes.
- Add the chloral hydrate and dissolve.
- Add the citric acid and mix well.
- Check that the pH is 2.45, adjust with citric acid if not.
- It may be used immediately.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 5-10 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
- The alcoholic hematoxylin solution should be aged (partially oxidised).
- The staining time was not given, but 5-10 minutes should suffice.
- 16 mL of a 10% solution of hematoxylin equates to 1.6 grams dye. With 60 grams of mordant and strong oxidation this would indicate a darkly staining progressive solution.
- 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
- Pusey’s modified Mayer’s hematoxylin,
Journal of Histotechnology, v.2, No.2, p.54, 1979
citing:
Villaneuva, A.R., (1976)
Methods of preparing and interpreting mineralized sections of bone, in:
Proceedings of the First Workshop on Bone Morphometry
Jaworski, Z. F. G., Editor, (1976).
