Krutsay's
Alum & Iron Hematoxylin

Krutsay's alum hematoxylin gives very highly selective staining of nuclei. It may also be easily converted into an iron hematoxylin for use as an acid resistant nuclear stain.

Alum hematoxylin	Amount	Function
Hematoxylin	1 g	Dye
Potassium alum	50 g	Mordant
Distilled water	1 L	Solvent
Hydrochloric acid	5 mL	Acidifier
Sodium iodate	0.2 g	Oxidant
Iron conversion	Amount	Function
Iron alum	2 g	Mordant
Hydrochloric acid	0.5 mL	Acidifer
Distilled water	100 mL	Solvent

 

Compounding procedure
Mix the reagents under Alum hematoxylin together.
Bring to the boil
Cool and filter.
The solution may be used immediately.

Conversion to iron hematoxylin
Mix the reagents under Iron conversion together, and dissolve.
For use mix the following:–
Alum hematoxylin solution – 25 mL
Iron conversion solution – 2 mL

Method

1. Bring sections to water with xylene and ethanol.
2. Place into either 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 (alum) or black (iron)
• Background  –  as counterstain or unstained

Notes

1. This is a progressive solution and is highly selective for nuclei.
2. The iron alum is (FeNH₄(SO₄)₂.12 H₂O)
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
Humason, G. L.
Animal Tissue Techniques.
W. H. Freeman and Co., San Francisco, CA, USA