[Histonet] Melanin Bleach

[Jeffrey Robinson]

Hi Karen-  I have also had problems getting tissue being stained for IHC markers to adhere to the slide.  The Potassium permanganate is really harse on the tissue (if you can get the sections to stay on).  Here is a trick I picked up at an NSH lecture that I have used successfully several times.

Run your IHC stains as usual.  Rinse well in water.  Do not counterstain with hematoxylin.  Stain with DiffQuik II for 30 seconds.  Rinse in water for a short period of time.  Differentiate with 10 dips each in 2 changes of 95% ETOH and 2 changes of 100% ETOH and then clear in xylene and coverslip.
Results:  melanin pigment will turn green.  Brown DAB stain will be unaffected.  Other tissue elements will be stained a medium blue color.
This method will also work for other Special Stains but I have not attempted to modify this method for use on "Red" stains.

Jeff Robinson, Senior Histotechnologist, Sierra Pathology Lab, Clovis, CA.

-----Original Message-----
From: Heckford, Karen - SMMC-SF via Histonet [mailto:histonet at lists.utsouthwestern.edu]
Sent: Tuesday, May 10, 2016 8:45 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Melanin Bleach

Good Morning,
I need some help.  Yesterday I bleached some heavily pigmented tissue.  I have to run some IHC's on them.   I bought the bleaching kit from American Master Tech.  I had to put the Permanganate for about 6 hours to get the melanin and then a couple of minutes in Oxalic Acid.  I had to let them set overnight because I could not get another IHC run in that day.   It looks like the tissue fell off during decloaking.    I used Apex slides.   I rarely ever have to bleach anything here.  So I am not sure if I did this correctly.   I am thinking I need to bleach and do the run in the same day and not let them set over night in DiH20.

Thanks,


Karen Heckford HT ASCP CE
Lead Histology Technician
St. Mary's Medical Center
450 Stanyan St.
San Francisco, Ca. 94117
415-668-1000 ext. 6167
karen.heckford at dignityhealth.org
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