Rawitz' Alum Hematoxylin Variants
Materials
| Material | Variant | Function | ||
|---|---|---|---|---|
| 1895a | 1895b | 1909 | ||
| Hematoxylin | 10 g | – | – | Dye |
| Hematein | – | 2.5 g | 2 g | Dye |
| Potassium alum | 10 g | – | – | Mordant |
| Ammonium alum | – | 15 g | – | Mordant |
| Aluminum nitrate | – | – | 20 g | Mordant |
| Distilled water | 650 mL | 500 mL | 500 mL | Solvent |
| Glycerol | 350 mL | 500 mL | 500 mL | Stabiliser |
Compounding Procedure
- Dissolve the aluminum salt and dye in the water.
- When dissolved, add glycerol.
- The 1895a formula should be allowed to ripen.
- The 1895b and 1909 formulas 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
- The 1895b and 1909 formulae use hematein instead of hematoxylin.
- The Microtomists Formulary and Guide gives the 1895b and 1909 formulae as containing 500 millilitres of glycerol. The Microtomists Vade-Mecum gives these two formulae as containing 500 grams of glycerol. This equates to 400 millilitres based on a specific gravity of 1.25.
- 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. - Bolles Lee, A.. Edited by Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist’s Vade-Mecum. 11 ed.,
Churchill, London, UK.
