Lillie's Alum Hematoxylin
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 5 g | Dye |
| Ammonium alum | 50 g | Mordant |
| Distilled water | 700 mL | Solvent |
| Glycerol | 300 mL | Stabilizer |
| Sodium iodate | 0.5 g | Oxidant |
| Glacial acetic acid | 20 mL | Acidifier |
Compounding Procedure
- Dissolve the dye and Alum in the water.
- Add the iodate, glycerol and acid.
- Leave overnight to ripen before using.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 5 minutes.
- 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
- This is a strong, regressive solution.
- 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
- Lillie, R.D.,
Histopathologic Technic and Practical Histochemistry
McGraw-Hill, New York, USA
