<html><div>Noelle,</div>
<br>
<div>did you really mean "........(10-20 dips in water then
acetone)....."? do you thoroughly dry your slides after the
water and before the acetone? i have always understood that acetone
contaminated with water could have an ill effect on immunoreactivity of
"some" antigens...........maybe not yours eh?</div>
<br>
<div>Rob</div>
<br>
<div>At 12:40 PM 5/28/99 -0400, you wrote:</div>
<div>>Tim,</div>
<div>></div>
<div>>I asked a similar question to histonet not too long ago. I
got enough ideas</div>
<div>>to run my own study. What I can tell you is that monkey
and human renal</div>
<div>>tissue are best when sectionned, fixed (I use Acetone at -20C),
air dried</div>
<div>>(about 15-30 min., the minimum to get dry slides), and stored at
-70C until</div>
<div>>use. When I remove the slides for IHC, I do a brief fix
(10-20 dips in</div>
<div>>water then acetone) for reasons based on what our staining
system needs.</div>
<div>>However, murine renal, pancreas, spleen, and lymph node tissue
do just fine</div>
<div>>if you fix in acetone, air dry, and store (dust-free) at room
temperature. </div>
<div>></div>
<div>>There are quite varied protocols to address your question.
Good luck. Feel</div>
<div>>free to ask ant nagging questions you may have about what I've
written.</div>
<div>></div>
<div>>Noelle Patterson</div>
<div>>Naval Medical Research Center</div>
<div>>Bethesda, Md</div>
<div>>pattersonn@NMRIPO.NMRI.NNMC.NAVY.MIL</div>
<div>></div>
<div>></div>
<div>><x-tab> </x-tab>----------</div>
<div>><x-tab> </x-tab>From: Coskran, Timothy M
[SMTP:timothy_m_coskran@groton.pfizer.com]</div>
<div>><x-tab> </x-tab>Sent: Thursday, May 27,
1999 3:34 PM</div>
<div>><x-tab> </x-tab>To:
'Histonet@pathology.swmed.edu'</div>
<div>><x-tab> </x-tab>Subject: Fixation of
frozen sections</div>
<div>></div>
<div>></div>
<div>><x-tab> </x-tab>Does anyone have any ideas on
whether it is better to cut frozens,</div>
<div>>fix and</div>
<div>><x-tab> </x-tab>store the sections or to cut
frozens, store, and then fix when your</div>
<div>>ready to</div>
<div>><x-tab> </x-tab>carry out a stain? We
currently cut frozen sections, dry briefly</div>
<div>>and then</div>
<div>><x-tab> </x-tab>store the slides at -80C until
we are ready to do an immuno stain.</div>
<div>>Then we</div>
<div>><x-tab> </x-tab>bring the slides to room temp,
fix and proceed with the stain. I've</div>
<div>>seem</div>
<div>><x-tab> </x-tab>some literature that suggests
fixing the slides immediately after</div>
<div>>they have</div>
<div>><x-tab> </x-tab>been sectioned and then storing
in PBS until ready to use.</div>
<div>></div>
<div>><x-tab> </x-tab>If anyone has a basic protocol
for frozen sections, would they be</div>
<div>>willing to</div>
<div>><x-tab> </x-tab>share?</div>
<div>></div>
<div>><x-tab> </x-tab>Thanks</div>
<div>></div>
<div>><x-tab> </x-tab>Tim Coskran</div>
<div>><x-tab> </x-tab>Pfizer</div>
>
<BR>
<font color="#0000FF"><i><div align="center">
Robert S. Geske<br>
Research Associate<br>
Center for Comparative Medicine and Department of Pediatrics<br>
Baylor College of Medicine</font></i></html>