[Histonet] melanoma IHC with frozen sections

From:Constance McManus

Greetings, all!

Is anyone out there doing Microphthalmia Transcription Factor (MiTF) IHC for melanocytes on frozen sections (Mohs surgery)?   I am doing skin, frozen sections at 3 um.  The problem is inconsistent staining. I embed 2 or 3 pieces of tissue in the same block, but one piece will not stain at all while the other piece[s] will stain beautifully.  This doesn't happen every time, and when it does, I can't see where i have done anything different from the protocol.   I have taken great care to be certain there is no overlapping OCT on the tissue sections, we put the sections in a 54 C oven for 4 minutes, then acetone fixation for 20 sec. before doing the staining.  

I am using NeoMarkers reagents:  
Ultra V blocker
One step  HRP polymer
Neomarkers MiTF

Does anyone know anything about using alk phos on skin?  Would there be any advantage in switching to AP ?

Connie McManus, HT
University of Utah
School of Medicine
Dept. of Dermatology

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