Anderson's
Iron Hematoxylin

Solution A	Amount	Function
Hematoxylin	0.5 g	Dye
100% ethanol	50 mL	Solvent
Distilled water	50 mL	Solvent
Calcium hypochlorite, 2% aqueous	5 mL	Oxidant
Solution B	Amount	Function
Ferric ammonium sulphate	3 g	Mordant
Distilled water	100 mL	Solvent
Sulphuric acid	2.5 mL	Acidifier

 

Compounding procedure
Make each solution seperately.
For use, add 2 volumes solution A to 1 volume solution B.
The working solution may be used immediately, but is not stable for long.

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 stock solutions are stable for some time.
2. The working solution should be made fresh.
3. It is claimed that the working solution rarely overstains, i.e. it is progressive.
4. Solution A incorporates calcium hypochlorite as an oxidising agent for hematoxylin. Presumably other oxidising agents would suffice equally as well. Sodium iodate (0.1 g or less) is the most common.
5. 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:–
    Anderson, J., (1929)
    How to stain the nervous system.
    Livingstone. Edinburgh, Scotland.

 

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