Mann's
Alum Hematoxylin

Ingredient	Amount	Function
Hematein	6 g	Dye
Potassium alum	35 g	Mordant
Distilled water	350 mL	Solvent
95% ethanol	320 mL	Solvent
Glycerol	250 mL	Stabiliser
Glacial acetic acid	30 mL	Acidifier

 

Compounding procedure
Dissolve the dye in the acetic acid.
Mix the ethanol and glycerol together, and add to the dye in acetic acid.
Dissolve the alum in the water, and add to the dye solution.
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. This solution uses hematein instead of hematoxylin.
2. The high dye content indicates this is a strong, regressive solution.
3. The staining time should be determined by trial.
4. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
5. 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.