[Histonet] frozen sections not adhering to slide

[ritta69.iwon]

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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