Hamilton
Alum Hematoxylin

This formula is based on Langeron's 1942 (Mayer's) formula. I have referred to it as Hamilton alum hematoxylin as I received it via the internet from a histotechnologist at the Hamilton Health Sciences Corporation, Hamilton, Ontario, Canada.

Ingredient	Amount	Function
Hematoxylin	2 g	Dye
Ammonium alum	75 g	Mordant
Distilled water	950 mL	Solvent
100% ethanol	50 mL	Solvent
Sodium iodate	0.3 g	Oxidant
Glacial acetic acid	30 mL	Acidifier

 

Compounding procedure
Dissolve the alum in the water.
Add the ethanol and mix well.
Add the hematoxylin and dissolve.
Add the sodium iodate and mix well.
Leave overnight in the dark to ripen.
Mix well, and add the acetic acid.
Filter.
The solution may be used immediately.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for 3 minutes.
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. Potassium alum may be used instead of ammonium alum.
2. The solution is progressive.
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
Personal internet communication, source unidentified.