[Histonet] frozen sections not adhering to slide

[Truscott, Tom]

I don't know where that attachment came from on the previous reply,. Anybody know?, but I wouldn't open it

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Truscott, Tom
Sent: Friday, November 20, 2009 2:09 PM
To: ritta69.iwon; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] frozen sections not adhering to slide

Hi, I think I would try rinsing them (after the antifreeze) in a 95% alcohol/5% PBS X3 before the next step (mounting on slides?) Tom Truscott

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of ritta69.iwon
Sent: Friday, November 20, 2009 10:19 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] frozen sections not adhering to slide


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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