Apathy's Alum Hematoxylin
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 3 g | Dye |
| Ammonium alum | 30 g | Mordant |
| Distilled water | 450 mL | Solvent |
| 95% ethanol | 250 mL | Dye solvent |
| Glycerol | 350 mL | Stabiliser |
| Glacial acetic acid | 10 mL | Acidifier |
| Salicylic acid | 0.3 g | Acidifier |
Compounding procedure
- Dissolve the dye in 100 mL water and 250 mL ethanol.
- Leave at room temperature to ripen (months).
- Dissolve the alum and acids in 350 mL water.
- Combine both solutions.
- Add glycerol.
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
- Lillie excludes the salicylic acid.
- 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
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co. - Lillie, R.D., (1954)
Histopathologic technique and practical histochemistry Ed.2
Blakiston, New York, USA.
