[Histonet] RE: Azure B as a counter-stain for melanin

Houston, Ronald Ronald.Houston <@t> nationwidechildrens.org
Fri Feb 11 11:25:32 CST 2011


we use a standard working Giemsa solution that is good for >1 month, stored at 4C. We automate the procedure on our Bonds: 3min Giemsa and 1 min Hematoxylin.

Ronnie Houston, MS HT(ASCP)QIHC
Anatomic Pathology Manager

ChildLab, a Division of Nationwide Children's Hospital


700 Children's Drive
Columbus, OH 43205
(P) 614-722-5450
(F) 614-722-2899
ronald.houston <@t> nationwidechildrens.org<mailto:ronald.houston <@t> nationwidechildrens.org>


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Margaryan, Naira
Sent: Friday, February 11, 2011 11:22 AM
To: histonet <@t> lists.utsouthwestern.edu; histonet-request <@t> lists.utsouthwestern.edu
Subject: [Histonet] Azure B as a counter-stain for melanin
Importance: High

Dear Histology Professionals,

I am using the Azure B as a counter-stain for melanin preparing solution by the protocol below. My questions are:

1. Is this solution should be fresh prepared every day?

2. How long I can store and use this solution?

3. Where this solution should be stored in the room or in the refrigerator?

My protocol is:

            Counterstain Method

      The use of Azure B as a counter-stain will stain melanin a blue green color.

            Azure B working reagent:

            4mL of a 25mg/mL aqueous solution of Azure B

                        3.4mL of 0.1M acetic acid

                        600uL of 0.1M sodium acetate

                        27mL of DI water

                        5mL acetone

                        Incubate for up to 30 minutes, followed by a 20-second hematoxylin counter-stain.  The use of either counterstain should give you good cytological detail, with blue-green melanin and brown DAB specific signal.

Thanks in advance,



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