Chemicals	Single	Double	Triple	Function
Ethylene glycol	250 ml	250 ml	250 ml	solvent
Distilled water	750 ml	750 ml	750 ml	solvent
Hematoxylin	2 g	4 g	6 g	dye
Sodium iodate	0.2 g	0.4 g	0.6 g	oxidizing agent
Aluminum sulfate	20 g	40 g	80 g	mordant
Glacial acetic acid	20 ml	20 ml	20 ml	pH control

Dissolve ethylene glycol in distilled water.

Add hematoxylin, then sodium iodate.

Add aluminum sulfate and acetic acid

Stir for 1 hour at room temperature.

Filter if necessary.

Staining Characteristic

Progressive

Suggested Use

Single strength for cytology.

Triple strength for immunohistochemistry counterstain (dilute 1:5) for 30 seconds and H&E (dilute 1:5) for 5 minutes.

Note

This recipe gives clear and sharp nucleus staining with little background. Good for both immunohistochemistry counterstain and H&E. This solution is also alcohol and mercury free so it is good for IHC counterstain when AEC substrate is used. When triple strength is used, dilute 1:5 with distilled water before use.