Régaud's
Iron Hematoxylin

Solution A	Amount	Function
Ferric ammonium sulphate	5 g	Mordant
Distilled water	100 mL	Solvent
Solution B	Amount	Function
Hematoxylin	1 g	Dye
90% ethanol	10 mL	Solvent
Distilled water	80 mL	Solvent
Glycerol	10 mL	Stabiliser
Solution C	Amount	Function
Potassium ferricyanide	2.5 g	Bleach
Sodium borate	2 g	pH control
Distilled water	100 mL	Solvent

 

Compounding procedure
Make each solution seperately.
Solutions A and B may be reused provided they are not contaminated by each other.
Solution C should not be reused.

Method

1. Bring sections to distilled water with xylene and ethanol.
2. Place into solution A at 50°C for 30 minutes.
3. Rinse well with distilled water.
4. Place into solution B at 50°C for 30 minutes.
5. Rinse well with distilled water.
6. Place into solution C until differentiated.
7. Wash in running tap water until blue.
8. Counterstain as required.
9. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  black
• Background  –  as counterstain or unstained

Notes

1. The solutions are stable for some time.
2. Solution C is Weigert's ferricyanide bleach and should be used fresh.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.