[Histonet] Melanin Bleaching

[Koehnlein, Melissa]

Melanin can be distinguished from DAB by counterstaining with Azure B, which gives a dark green color to the melanin.  We've used the procedure below with success:

Immunoperoxidase technique modified by counterstain with azure B as a diagnostic aid in evaluating heavily pigmented melanocytic neoplasms:
http://onlinelibrary.wiley.com/doi/10.1111/j.1600-0560.1991.tb01381.x/abstract

Melissa Koehnlein
Lab Technician
Prairie Diagnostic Services Inc.
52 Campus Drive, Saskatoon, SK S7N 5B4
Phone (306) 966-7223
Fax (306) 966-2488
Email mailto:melissa.koehnlein <@t> pds.usask.ca

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Today's Topics:

   1. Melanin Bleaching (Janci Wellborn)
   2. Re: Melanin Bleaching (Rene J Buesa)


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Message: 1
Date: Wed, 26 Dec 2012 15:39:40 -0600
From: "Janci Wellborn" <JWellborn <@t> bjc.org>
Subject: [Histonet] Melanin Bleaching
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <50DB1A3C020000B2000E79E3 <@t> upbcgwg06>
Content-Type: text/plain; charset="us-ascii"

I am looking for alternatives to the potassium permanganate melanin bleaching procedure when doing IHC. I have seen literature references to doing a 3% H2O2 at 55 C after the IHC staining. I have also done a Giemsa counterstain in the past. Does any one have either procedure that they would be willing to share?

We are a small volume lab and the costs associated with bringing in a different color IHC kit is not justified.

Any help would be greatly appreciated.

Janci



Janci R Wellborn, HTL (ASCP)
Laboratory Technical Coordinator
Histology
Alton Memorial Hospital
1 Memorial Drive
Alton, Ill 62002
618-463-7082

------------------------------

Message: 2
Date: Wed, 26 Dec 2012 13:47:23 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Melanin Bleaching
To: Janci Wellborn <JWellborn <@t> bjc.org>,
        "histonet <@t> lists.utsouthwestern.edu"
        <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <1356558443.97578.YahooMailNeo <@t> web163106.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Melanin bleaching has to be done prior to the IHC procedure and the most reliable?way is using potassium permanganate. Any other option will produce less than optimal results.
Ren? J.

From: Janci Wellborn <JWellborn <@t> bjc.org>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Wednesday, December 26, 2012 4:39 PM
Subject: [Histonet] Melanin Bleaching

I am looking for alternatives to the potassium permanganate melanin bleaching procedure when doing IHC. I have seen literature references to doing a 3% H2O2 at 55 C after the IHC staining. I have also done a Giemsa counterstain in the past. Does any one have either procedure that they would be willing to share?

We are a small volume lab and the costs associated with bringing in a different color IHC kit is not justified.

Any help would be greatly appreciated.

Janci



Janci R Wellborn, HTL (ASCP)
Laboratory Technical Coordinator
Histology
Alton Memorial Hospital
1 Memorial Drive
Alton, Ill 62002
618-463-7082

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