Lillie & Earle's Iron Hematoxylin
Materials
Solution A
| Material | Amount | Function |
|---|---|---|
| Hematoxylin | 1 g | Dye |
| 95% ethanol | 50 mL | Solvent |
| Glycerol | 50 mL | Solvent |
Solution B
| Material | Amount | Function |
|---|---|---|
| Ferric ammonium sulfate | 15 g | Mordant |
| Ferrous sulfate | 15 g | Mordant |
| Distilled water | 100 mL | Solvent |
Compounding Procedure
- Make each solution separately.
- For use, combine equal parts of solutions A and B.
- The working solution may be used immediately.
Protocol
- Bring sections to water with xylene and ethanol.
- Place into the staining solution for an appropriate time.
- Rinse with tap water.
- If overstained, 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
- The stock solutions are stable for some time.
- The life of the working solution is not given. It is likely not stable for long, although the inclusion of glycerol and ferrous sulfate may improve its stability.
- The staining time should be determined by trial, but 10-30 minutes should suffice.
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:
Lillie and Earle, (1937).
American Journal of Pathology, v. 15, p. 765.
