[Histonet] RE: Cresyl violet stain on 50 um mouse brain sections

[Houston, Ronald]

Susan,
If you are talking about frozen sections of fixed tissue, they do not adhere well to slides (even charged slides) no matter how thick or thin. I'm not sure why you are treating the sections with alcohol chloroform either. That is pretty harsh treatment and totally unnecessary for Nissl staining. 

Why not stain the sections free-floating and then mount them from xylene?

Ronnie Houston
Anatomic Pathology Manager
Nationwide Children's Hospital
Columbus OH 43205
(614) 722 5450

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Truscott, Tom
Sent: Friday, July 29, 2011 10:22 AM
To: Michael, Susan; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Cresyl violet stain on 50 um mouse brain sections

Hi Susan, I would suggest cutting lots thinner sections. The plus slides seem to have trouble holding on to very thick sections. Another possibility is too much glacial acetic acid in your working solution of cresyl-too much acid can loosen the bonds on the plus slides. Tom T

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Michael, Susan
Sent: Friday, July 29, 2011 6:12 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Cresyl violet stain on 50 um mouse brain sections

I am having trouble keeping my sections on the slides when I stain with cresyl violet.  These are fixed frozen sections, 50 um, dried overnight on plus slides, overnight in 1 to 1 alcohol/chloroform, then rehydrated through 100, 95 ETOHs then to water.  Into the cresyl violet solution, then 100% to 95% ETOH.  Is it the water?  Is it the slides?  Any suggestions?

Susan
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