Lillie & Earle's
Iron Hematoxylin

Solution A	Amount	Function
Hematoxylin	1 g	Dye
95% ethanol	50 mL	Solvent
Glycerol	50 mL	Solvent
Solution B	Amount	Function
Ferric ammonium sulphate	15 g	Mordant
Ferrous sulphate	15 g	Mordant
Distilled water	100 mL	Solvent

 

Compounding procedure
Make each solution seperately.
For use, combine equal parts of solutions A and B.
The working solution may be used immediately.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for an appropriate time.
3. Rinse with tap water.
4. If overstained, dip briefly in 1% hydrochloric acid in 70% ethanol.
5. Wash well in running tap water to blue.
6. Rinse with distilled water.
7. Counterstain if desired.
8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  black
• Background  –  as counterstain or unstained

Notes

1. The stock solutions are stable for some time.
2. The life of the working solution is not given. It is likely not stable for long, although the inclusion of glycerol and ferrous sulphate may improve its stability.
3. The staining time should be determined by trial, but 10-30 minutes should suffice.

 

Reference

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.