Janssen's
Iron Hematoxylin

Ingredient	Amount	Function
Hematoxylin	1 g	Dye
Ferric ammonium sulphate	5 g	Mordant
100% ethanol	5 mL	Solvent
Distilled water	70 mL	Solvent
Glycerol	15 mL	Solvent
Methanol	15 mL	Solvent

 

Compounding procedure
Gray
Dissolve the iron alum into the water.
Dissolve the hematoxylin into the ethanol.
Combine, and leave for one week at room temperature.
Filter, and then add the other ingredients.
 
Lillie
Combines all solutions immediately they are dissolved,
and omits the week at room temperature.
 
The solution is stable for a few months.

Method

1. Bring sections to water with xylene and ethanol.
2. Place into the staining solution for 15-30 minutes.
3. Rinse with tap water.
4. Dip briefly in 1% hydrochloric acid in 70% ethanol.
5. Wash well in running tap water to blue.
6. Rinse with distilled water.
7. Counterstain if desired.
8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  black
• Background  –  as counterstain or unstained

Notes

1. This solution may be used as an acid resistant nuclear stain.
2. It is recommended as a substitute for Weigert's iron hematoxylin.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Janssen, (1897), Cellule, v.14, p.207.
    Lillie & Earle, (1939), Stain Technology, v.14, p.53