Langeron's Alum Hematoxylin Variants
Both of Langeron’s formulae are modifications of Mayer’s formulae. Langeron’s 1924 formula is a modification of Mayer’s 1896 formula, and Langeron’s 1942 formula is a modification of Mayer’s 1901 formula.
Langeron’s 1942 formula is the solution usually meant when Mayer’s hemalum is specified.
Materials
| Material | Solution | Function | |
|---|---|---|---|
| 1924 | 1942 | ||
| Hematoxylin | 4 g | 1 g | Dye |
| Ammonium alum | 50 g | – | Mordant |
| Potassium alum | – | 50 g | Mordant |
| Distilled water | 700 mL | 1 L | Solvent |
| Glycerol | 300 mL | – | Stabilizer |
| Sodium iodate | – | 0.2 g | Oxidant |
| Glacial acetic acid | 20 mL | – | Acidifier |
| Citric acid | – | 1 g | Acidifier |
| Chloral hydrate | – | 50 g | Acidifier |
Compounding Procedures
1924
- Grind the hematoxylin with glycerol in a pestle and mortar.
- Dissolve the other ingredients in water.
- Add the hematoxylin paste.
- Wash out the hematoxylin paste with the solution.
- Leave months to ripen.
1942
- Dissolve the Alum and dye in the water.
- Add the other ingredients.
- Bring to the boil.
- Cool to room temperature.
- The solution may be used immediately.
- Ammonium Alum may be substituted for potassium alum.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for an appropriate time.
- Rinse well with water.
- Differentiate the 1924 solution 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 1924 solution is regressive, and staining time should be determined by trial.
- The 1942 solution is progressive. Staining time is 5-10 minutes.
- 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.
