[Histonet] Frozen sections falling off of slides

Jacqueline D. Garfield JGarfield <@t> lifecell.com
Tue Apr 3 10:13:36 CDT 2007

Hello Dawn,

We occasionally have a similar problem with our paraffin sections for
IHC.  I haven't heard of "Halt solution".  Do you use it for paraffin
sections?  If so where can I purchase it?  

Thank you,

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Dawn
Sent: Tuesday, April 03, 2007 10:39 AM
To: Stacey Barrick; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] Frozen sections falling off of slides

hi Stacey,
  Have you tried using Halt solution in your waterbath? We use a capful
in each waterbath. It does not interfere with immunostaining like some
of the other adhesives do.
  It might be worth a try.

Stacey Barrick <barrickstacey <@t> yahoo.com> wrote:
  Hi everyone, 

I'm cutting frozen rat scg at 10um in OCT. I cannot keep the sections
from falling off the slides during the overnight incubation in primary
anitbody. I have tried the Superfrost plus slides and currently we are
using the Superfrost Plus GOLD slides which are supposed to be even
better. THis is what I've tried so far:
Slides on warmer for 1hr
Slides on warmer for 2hr
Slides on warmer for 5hr
Slides on warmer for 2hr, then at RT overnight

Sometimes ALL sections fall off. Other times just some of the sections
will fall off. 

When I remove them from the warmer, I cover them with PBS to rehydrate.
THen incubate overnight in my primary antibody at 4deg C.

The next morning is when I notice all of the floaters. THey are usually
foggy when the come out of the fridge. When I put the slides into PBS
most if not all of the sections are gone. 

Please help! This is precious tissue and I can't loose anymore. I've
never had this problem with any other tissue I've used. I dont believe
its been a problem in the past in this lab either but noone knows what
the difference is between then and now.

Also, what setting do you use on the slide warmer (temp?) I've tried
setting it between 2 and 3 (usually 40deg C)

Thanks in advance!!


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