Re: Renal biopsies

From:Richard Cartun (by way of Histonet)

All of our renal biopsies are received fresh and I freeze the portion
for immunofluorescence in our cryostat (-20 to -22 degrees C.).  I have
never noticed a problem with our freezing.  Our immunofluorescence
preparations are beautiful!

Richard Cartun
Director, Immunopathology
Hartford Hospital

>>> Lisa Black  07/16/03 02:52PM >>>
Hello All,

I would like to have information from other sites regarding the way
renal biopsies are handled.

At the lab where I work, the bx is received in Zeus fixative
(Michel's).  It is then placed in wash solution for 30 minutes.  At
that
time a thin film of OCT is placed on a cork square and the bx is
placed
on the cork.  The cork with the bx is then placed in a vial of -70#186#
C
chilled isopentane and stored in the -70#186#C freezer until time to
cut.

A thin film of OCT is placed onto a cryostat chuck.  Before it freezes
completely, the specimen is obtained from the freezer and the cork is
attached.  Sections are cut on the cryostat at 3 microns with 1
attached
to each slide.  15 slides are cut and allowed to dry for one hour
prior
to staining.

This is the method of choice of the renal pathologist.  The theory is
to decrease freeze artifact.  Using this method causes great
difficulty
in cutting a tiny specimen without any surronding OCT.  I have techs
that have worked at other sites that say there are much easier and
more
efficient ways to perform this task.  Would those of you who routinely
cut renals be willing to give me advice?  Any information would be
greatly appreciated.

Thanks in advance.

Lisa Black
Carilion Consolidated Lab
540-985-4082
LBLACK@CARILION.COM





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