Sass' Alum Hematoxylin Variants
Materials
| Material | Variation I | Variation II | Function |
|---|---|---|---|
| Hematoxylin | 10 g | 1 g | Dye |
| Ammonium alum | saturated | 50 g | Mordant |
| Distilled water | 1 L | 1 L | Solvent |
| Sodium iodate | 10 g | 1 g | Oxidant |
| Glacial acetic acid | 30 mL | – | Acidifier |
Compounding Procedure
- Dissolve the Alum in the water.
- Add the hematoxylin.
- Add the other ingredients.
- Filter.
- The solutions may be used immediately.
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
- Both of these formulae use 1 gram of sodium iodate for each gram of hematoxylin. However, 1 gram of hematoxylin is oxidized by 0.2 grams of sodium iodate, so the solutions should be expected to have a relatively short life.
- Due to it’s high dye content, the Var I formula is likely regressive.
- The Var II formula is likely progressive.
- The 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.
