Janssen's Iron Hematoxylin
Materials
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 1 g | Dye |
| Ferric ammonium sulfate | 5 g | Mordant |
| 100% ethanol | 5 mL | Solvent |
| Distilled water | 70 mL | Solvent |
| Glycerol | 15 mL | Solvent |
| Methanol | 15 mL | Solvent |
Compounding Procedure
Gray Method
- Dissolve the iron alum into the water.
- Dissolve the hematoxylin into the ethanol.
- Combine, and leave for one week at room temperature.
- Filter, and then add the other ingredients.
Lillie Method
- Combines all solutions immediately once they are dissolved, and omits the week at room temperature.
- The solution is stable for a few months.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for 15-30 minutes.
- Rinse with tap water.
- Dip briefly in 1% hydrochloric acid in 70% ethanol.
- Wash well in running tap water to blue.
- Rinse with distilled water.
- Counterstain if desired.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Background – as counterstain or unstained
Notes
- This solution may be used as an acid resistant nuclear stain.
- It is recommended as a substitute for Weigert’s iron hematoxylin.
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.
Citing:
Janssen, (1897),
Cellule, v.14, p.207.
Lillie & Earle, (1939),
Stain Technology, v.14, p.53
