[Histonet] melanin counterstaining

Margaryan, Naira NMargaryan <@t> childrensmemorial.org
Wed Aug 17 13:51:23 CDT 2011

Good morning,

I have heavily pigmented melanocytic cells and trying to differentiate between melanin pigment and chromogen (DAB) by using Azure B.

After IHC with DAB I incubate slides for 30-45 min in the working solution of:

 *   4ml of a 25mg/ml aqueous solution of Azure B
 *   3.4ml of 0.1M acetic acid
 *   600µl of 0.1M sodium acetate
 *   27ml of DI water
 *   5ml acetone

Then water wash and hematoxylin counter-stain. By the end all melanin supposed to turn blue-green, but I still see some remained brown melanin.

My question is: I would like to increase green color, so the concentration of what reagent I have to increase?

Thanks in advance,

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