[Histonet] frozen special stains

From:"Ingles Claire"



Hello out there!
I have a question for the collective histo-brain out there. (resistance is futile). Does anyone do special stains i.e. trichrome and/or elastic stains on frozen tissue (skin)? I have been trying these out using the regular protocols you would use for FFPE tissues. The (FFPE) controls I stain with the frozens come out fine, but the staining of the frozen sections is so intense that it is difficult to interpret. On the trichrome, the epidermis is so dark it looks black.  I've tried cutting the incubation times back by as much as half. It helps a bit but is still pretty intense. Are there any special procedures out there to make them turn out more normal looking? For procedure, I cut sections at 5 microns, dry in a 60 oven for 10 min, and fix in 10% formalin for 30 min. Then rinse in running tap for about 5 minutes and perform stain as normal. Help?!?
 
Claire Ingles
UW Hospital
Madison WI

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