Gill's Hematoxylin Protocol




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




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.




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.