[Histonet] frozen sections not adhering to slide
ritta69.iwon
ritta69 <@t> iwon.com
Fri Nov 20 12:19:28 CST 2009
I am processing adult rat brain sections for routine histological stains, but am having a terrible time getting them to stick to the slide. After transcardial perfusion the rat brains are dissected out and processed for frozen sectioning. I post-fix overnight in PFA, cryoprotect in 30%sucrose until the brains sink and flash freeze in isopentane. All seems like pretty standard stuff. After cutting 45 um thick sections and storing them in antifreeze solution (30% 0.1M Phosphate Buffer, 30% Glycerol, 40% Ethylene Glycol), I mount them on slides. After letting them dry overnight, I rehydrate/wash in 0.1M Phosphate Buffer. Frequently, the sections start to wrinkle up and start coming of the slide during this first step. I'm using superfrost+ slides from fisher. Does anyone have any advice? Please help.
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