[Histonet] Melanin Bleach

[Tony Henwood (SCHN)]

The KMnO4/oxalic acid sequence will weaken the tissue adherence.
If you do not have to use Peroxidase/DAB, then I would recommend alkaline phosphatase labelling (+red chromagen).

Remember KMnO4/oxalic acid can be deleterious to some antigen-antibody reactions. 
We found that LCA, CAM5.2, L26, alpha-fetoprotein, and NSE were labile, whilst alpha-1-antitrypsin, HBsAg, CMV, CEA, Thyroglobin and chromogranin showed decreased staining. EMA, desmin, HSVII, S100, GFAP, PSA, PAcP, Calcitonin, ACTH and Prolactin were found to be resistant to permanganate treatment.

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 


-----Original Message-----
From: Heckford, Karen - SMMC-SF via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Wednesday, 11 May 2016 1:45 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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