Bosma's
Alum Hematoxylin

Ingredient	Amount	Function
Hematoxylin, 10% alc.	25 mL	Dye
Ammonium alum	25 g	Mordant
Tap water	865 mL	Solvent
Diethylene glycol	100 mL	Stabiliser
Sodium iodate	0.2 g	Oxidant
Glacial acetic acid	10 mL	Acidifier

 

Compounding procedure
Place 815 mL hot tap water in a large flask.
Add the ammonium alum, mix well to dissolve and cool to room temperature.
Add the alcoholic hematoxylin solution (ripened).
Add the sodium iodate dissolved in 50 mL cold tap water and mix well.
Add the diethylene glycol and mix well.
Add the acetic acid and mix well.
The pH should be 3.1 to 3.3 and the 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 well with water.
4. Differentiate with acid ethanol if necessary.
5. Rinse with water and blue.
6. Rinse well with water.
7. Counterstain if desired.
8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  blue
• Background  –  as counterstain or unstained

Notes

1. The staining time should be determined by trial.
2. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
3. Blueing is done with alkaline solutions such as hard tap water, Scott’s tap water substitute, 0.1% ammonia water, 1% aqueous sodium acetate, 0.5% aqueous lithium carbonate etc.

 

Reference
Bosma, Rob
A useful hematoxylin without toxic chemicals.
Histologic, V 18, Nº 1, January 1988