|Ethylene glycol||250 ml||solvent|
|Distilled water||750 ml||solvent|
|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.
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.