[Histonet] Question/Help on apoptosis assay

From:Rich Ehrlichman

Hello all,  I am new to this list and also new to histology in general.  This list seems like a very useful tool for anyone doing histology so I thought I'd give it a shot. I'm a student working in a lab at U Penn.  We are currently working on staining for apoptosis in mouse brains using the Promega Tunel kit.  They are fixed for 24 hours in 10% buffered formalin and we have been trying to use fresh vibratomed sections cut at 50 um.  We are having a problem with the sections coming off of the slides during the assays.  They wouldn't stick to slides we had gelled ourselves so on a suggestion from someone with more experience we purchased ultra stick slides.  These didn't work any better.  We basically floated the sections, let them sit in a humidified chamber for about 5 minutes and then started the assay.  It has been suggested that we should let them sit over night in the humidified chamber or possibly use heat to get them to stick.  I'm having trouble finding out whether that would
 be a problem for the tissue since these are not in paraffin.  So basically my question is, does anyone have any tips or experiences with getting the sections to stay put or just general methods we can try that wont compromise the tissue?  Thank you for any comments/help.
 
Rich




Siegel Lab
Clinical Research Building
University of Pennsylvania
		
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