[Histonet] RE: melanin counterstaining

Diane.Craft <@t> amcny.org Diane.Craft <@t> amcny.org
Wed Aug 17 14:46:43 CDT 2011

I am not sure what others do, but I do not run my Azure B slides through 
alcohol because I find the alcohol diminishes the intensity of the green. 
I air dry, clear, then coverslip.

Diane Craft
Pathology Department
Animal Medical Center
510 East 62nd St
New York NY 10065-8314
212-329-8675 (phone)
212-759-5878 (fax)

Steve Olsen <solsen <@t> MPLNet.com> 
Sent by: histonet-bounces <@t> lists.utsouthwestern.edu
08/17/2011 02:56 PM

"Margaryan, Naira" <NMargaryan <@t> childrensmemorial.org>, 
"histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>, 
"histonet-request <@t> lists.utsouthwestern.edu" 
<histonet-request <@t> lists.utsouthwestern.edu>

[Histonet] RE: melanin counterstaining

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

P  865.380.9746 ext.187
C  865.978.0321
F  865.380.9191


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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [
mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Margaryan, 
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 

*   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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