RE: Fixation of frozen sections

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From:Rob Geske <>
To:"Patterson, Noelle" <PattersonN@NMRIPO.NMRI.NNMC.NAVY.MIL>
Date:Fri, 28 May 1999 15:10:18 -0500
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<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 []</div> <div>><x-tab>   </x-tab>Sent:  Thursday, May 27, 1999 3:34 PM</div> <div>><x-tab>   </x-tab>To:  ''</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>
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