Lillie's
Alum Hematoxylin

Ingredient	Amount	Function
Hematoxylin	5 g	Dye
Ammonium alum	50 g	Mordant
Distilled water	700 mL	Solvent
Glycerol	300 mL	Stabiliser
Sodium iodate	0.5 g	Oxidant
Glacial acetic acid	20 mL	Acidifier

 

Compounding procedure
Dissolve the dye and alum in the water.
Add the iodate, glycerol and acid.
Leave overnight to ripen before using.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for 5 minutes.
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 is a strong, regressive solution.
2. The staining time should be determined by trial.
3. Acid ethanol is 0.5% - 1% hydrochloric acid in 70% ethanol.
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
Lillie, R.D.,
Histopathologic Technic and Practical Histochemistry
McGraw-Hill, New York, USA