RE: Renal biopsies

From:"Gutierrez, Juan"

Hello Lisa, at the place where I used to work recently, we used cryomolds to
freeze our renal biopsies.  They are real simple to use and they offer great
results.  Like you, we would receive the biopsies in Michel's solution, but
instead of tris we used maleimide buffer for 15 min.  We used to get it from
Poly Scientific R&D Corp. 1-800-645-5825.
After washing in buffer, we would pat dry the bx on a piece of filter paper,
just enough to remove the excess fluid.  We would then lay it down at the
bottom of the intermediate size cryomold (I believe Sakura makes them) and
then fill the mold with OCT.  We would then float the mold on a histobath
until frozen solid.  The key to floating the tissue is that you get a nice
straight surface to cut when the block freezes.  If you let the mold sink in
the isopentane, the mold will bow out in the center giving you an uneven
surface to cut.  
If you need to store the tissue, you can leave it in the mold as they are
disposable.  I would first place them in a baggie to prevent drying.
Hope this helps,

Juan C. Gutierrez, HT(ASCP)
Histology Supervisor
Christus Santa Rosa Hospital

-----Original Message-----
From: Lisa Black []
Sent: Wednesday, July 16, 2003 1:53 PM
Subject: Renal biopsies

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

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