July 15, 2016

Gill’s Hematoxylin Solution

Ingredient 1x Function
Ethylene glycol  250 ml solvent
Distilled water  750 ml solvent
Hematoxylin 2 g dye
Sodium iodate 0.2 g oxidizer
Aluminum sulphate 20 g mordant
Glacial acetic acid 20 ml acidifier


Dissolve ethylene glycol in distilled water.
Add hematoxylin, then sodium iodate.
Add aluminum sulphate and acetic acid
Stir for 1 hour at room temperature.
Filter if necessary.

Suggested Use:
1. One minute for H&E staining with periodic agitation.
2. Dilute 1/2x (in H2O) as an IHC counterstain.

This recipe gives clear and sharp nucleus staining with little background. It the blue component of an H&E stain and stains nucleic acids (i.e. the nucleus) blue. It also is useful as an immunohistochemistry counterstain. Unlike Harris' Hematoxylin this solution is mercury free so it is good for IHC counterstain when AEC substrate is used. More concentrated solutions can be made by proportionally increasing all of the ingredients conentrations (except for the solvents).

For everything you wanted to know about hematoxylin but were afraid to ask, see the fascinating note by Gary Gill in the Histonet Archives and Hematoxylin Technical Notes in the Stains File website.

Tags: Fero-Lab-Protocols