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

[Montina Van Meter]

Susan,

I have stained 60um fixed sections on plus slides without any problem. I
free float the sections in a multi-well plate containing PBS or TBS.
When mounting the sections onto the plus slide, I wipe away the excess
fluid after EACH section, so that the previous section will not be
exposed to the PBS again which results in unequal drying. I also place
the slide in a slanted position on the slide tray and allow them to dry
overnight.  The next morning you could also dry them in a 37 degree oven
for a few minutes to help in adherence. 

I have had others in the lab mount sections at the same time as I do and
have often observed that their sections will flap on the slide during
staining. Mine, on the other hand, don't come off of the slide. I can
only surmise that this is due to many years of experience in mounting
sections and the importance of "wicking" away the excess buffer.

Good luck,

Tina



-----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 8:12 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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