Lugol’s Hematoxylin

Iodine	Amount	Function
Iodine	2 g	Oxidant
Potassium iodide	4 g
Distilled water	100 mL	Solvent
Hematoxylin	Amount	Function
95% ethanol	100 mL	Solvent
Hematoxylin	10 g	Dye
Alum	Amount	Function
Potassium alum	6 g	Mordant
Distilled water	500 mL	Solvent

Working solution	Amount
Hematoxylin	1.5 mL
Lugol’s iodine	1.5 mL
Alum	100 mL

 

Compounding procedure
Make each solution as listed.
Combine as directed above.
Leave 15 minutes before use.

Method

1. Bring sections to distilled water with xylene and ethanol.
2. Place into the staining solution for 5-10 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. Read these comments on Lugol’s iodine.
2. The solution is usable for a minimum of one week.
3. The working solution will stain at least 100 slides.
4. Since the solution is not acidified, a distilled water rinse before staining will reduce alkaline carry over.
5. The authors named this solution Lugol’s Hematoxylin.
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
Clarke, G. and Dodds, H. M., (1983)
Lugol’s Hematoxylin.
Stain Technology, v 58, Nº 4, p. 232