Carazzi's
Alum Hematoxylin

Ingredient	Amount	Function
Hematoxylin	1 g	Dye
Potassium alum	50 g	Mordant
Distilled water	800 mL	Solvent
Glycerol	200 mL	Stabiliser
Sodium iodate	0.2 g	Oxidant

 

Compounding procedure
Dissolve the hematoxylin in the glycerol.
Dissolve the alum in 750 mL of the water.
Dissolve the sodium iodate in the remaining 50 mL water.
Add the alum solution to the hematoxylin solution slowly, while mixing well.
Add the sodium iodate solution. Mix well.
Filter.
The solution may be used immediately, and is stable for about six months.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for 5 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. This is a progressive solution giving litttle background staining.
2. Doubling the hematoxylin to 2 g. intensifies nuclear staining.
3. The double strength solution is recommended for frozen sections with about 1 minute staining time.
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
Bancroft, J.D. and Stevens A. (1982)
Theory and practice of histological techniques, Ed. 2
Churchill Livingstone, Edinburgh & London, UK.