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
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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