Papamiltiades'
Alum Hematoxylin

Ingredient	Amount	Function
Hematoxylin	4 g	Dye
Aluminum sulphate	10 g	Mordant
Distilled water	900 mL	Solvent
Zinc sulphate	5 g	–
Potassium iodide	4 g	Stabiliser
Glacial acetic acid	32 mL	Acidifier
Glycerol	100 mL	Stabiliser

 

Compounding procedure
Dissolve the hematoxylin in 400 mL water.
Dissolve the aluminum sulphate in 200 mL water.
Dissolve the zinc sulphate in 100 mL water.
Dissolve the potassium iodide in 100 mL water.
Combine the four solutions, then add the acetic acid and glycerol.
The solution may be used immediately, and is stable for about two months.

Method

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

Expected results

• Nuclei  –  blue
• Background  –  as counterstain or unstained

Notes

1. The solution is progressive.
2. The purpose of the zinc sulphate and potassium iodide are not clear.
3. The staining time should be determined by trial.
4. 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
Putt, F.A.
Manual of Histopathological Staining Methods
John Wiley & Sons, New York, NY., USA