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Gill’s Hematoxylin Protocol
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Gill’s Hematoxylin Protocol
Chemicals  Single  Double  TripleFunction  
Ethylene glycol  250 ml  250 ml  250 ml  solvent  
Distilled water750 ml750 ml750 ml  solvent
Hematoxylin2 g4 g  6 g  dye
Sodium iodate0.2 g0.4 g0.6 g  oxidizing agent
Aluminum sulfate  20 g  40 g  80 g  mordant  
Glacial acetic acid20 ml20 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.

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