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Alum Hematoxylin Variants

Ingredient	Var I	Var II	Function
Hematoxylin	10 g	1 g	Dye
Ammonium alum	saturated	50 g	Mordant
Distilled water	1 L	1 L	Solvent
Sodium iodate	10 g	1 g	Oxidant
Glacial acetic acid	30 mL	–	Acidifier

 

Compounding procedure
Dissolve the alum in the water.
Add the hematoxylin.
Add the other ingredients.
Filter.
The solutions 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. Both of these formulae use 1 gram sodium iodate for each gram hematoxylin. However, 1 gram hematoxylin is oxidised by 0.2 grams sodium iodate, so the solutions should be expected to have a relatively short life.
2. Due to it's high dye content, the Var I formula is likely regressive.
3. The Var II formula is likely progressive.
4. The staining time should be determined by trial.
5. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
6. 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
Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.