RE: Any tips for "no distortion" fixation and cutting of lung?

From:"Sebree Linda A."

Hi fellow UWHCer,

Have you considered the gentle Jane freezing and tape transfer method put out by Instrumedics?  I used to use it on cross sections of rabbit heart. I needed adjacent sections as near identical as possible to compare histochemical staining and autoradiography. I believe I fixed my sections in acetone.

If you need more info., you know where to find me.


Linda A. Sebree, HT(ASCP)
University of Wisconsin Hospital & Clinics
IHC/ISH Laboratory
600 Highland Ave.
Madison, WI 53792
FAX: (608)262-7174

-----Original Message-----
From: LaCinda Burchell []
Sent: Wednesday, March 26, 2003 10:15 AM
Subject: Any tips for "no distortion" fixation and cutting of lung?

Hello Histonet,
  Investigators here wish to obtain sections of rat lung tissues with
no (or very  minimal distortion) of rat lung representing it's original
living state.  IE. no shrinkage, compression, or fractures.  We've tried
several variations on cryoprotectants freezings and such, but have yet
to obtain "perfect" results.  Paraffin processing is out due to
shrinkage in processing and expansion on the waterbath.  They want to be
able to see small airways within the context of the whole rat lung so EM
seems to be out of the question.  I won't be asked to do IHC on these
tissues so overfixation isn't an issue.  Prefusion is out of the
question because it causes pressure changes and distortions of the
tissue.  I'm currently considering traditional fixation techniques
(formalin?) followed by cryosectioning.   Any suggestions or insights
would be very appreciated.  Bless you if you can offer any help!   Cindy

LaCinda Burchell, BA, AS, HT(ASCP)
University of Wisconsin-Madison, Medical School
Asthma and Allergy Research IHC Lab
600 Highland Ave.  CSC  K4/913
Madison, Wisconsin  53792

Phone: 608-262-3518
FAX:     608-263-3746

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