[Histonet] RE: melanin counterstaining

Steve Olsen solsen <@t> MPLNet.com
Wed Aug 17 13:56:51 CDT 2011

You can try a simple Schmorl's stain - which stains melanin.  You can find it in a Sheehan/Shrapchek

Steven K. Olsen
Chief Operating Officer, Director of Anatomic Pathology
Molecular Pathology Laboratory Network, Inc.
250 E. Broadway, Maryville, TN 37804

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira
Sent: Wednesday, August 17, 2011 2:51 PM
To: histonet <@t> lists.utsouthwestern.edu; histonet-request <@t> lists.utsouthwestern.edu
Subject: [Histonet] melanin counterstaining

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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