Thomas'
Iron Hematoxylin

Solution A	Amount	Function
Hematoxylin	2.5 g	Dye
Dioxane	40 mL	Solvent
Hydrogen peroxide	1 mL	oxidiser
Solution B	Amount	Function
Ferric ammonium sulphate	3 g	Mordant
Ferric chloride	1.5 g	Mordant
Ferrous chloride	6 g	Mordant
Distilled water	60 mL	Solvent
Acetic acid	6 mL	Acidifier

 

Compounding procedure
Dissolve the hematoxylin into the dioxane.
Add the peroxide.
Add the acetic acid to the water.
Dissolve the iron salts into the dilute acetic acid.
Filter the solution of iron salts into the hematoxylin solution.

Method

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

Expected results

• Nuclei  –  black
• Background  –  as counterstain or unstained

Notes

1. The solution is likely stable for some time.
2. Dioxane is no longer used in histotechnology for safety reasons. The method should be considered obsolete.
3. The solution is progressive, and the staining time should be determined by trial.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.  Citing:–
    Thomas, (1943)
    Bulletin d'histologie appliquée à la physiologie et à la pathologie
     et de technique microscopique, v. 20, p. 212.