RE: Renal biopsies

From:Rena Fail

   Renal biopsies are received in Michel's fixative, washed in 3 changes
of Tris buffered saline 5 minutes each. We have drilled holes in the
upper sides of a metal mold and attached a wire loop. A rectangle of
foil( pt number written on both sides with a cryopen) is molded to the
bottom of the mold using a pencil eraser. Oct is placed in the mold up
to the edge of the small square, not up to the level you would fill with
parrafin. The tissue is oriented in the Oct on the bottom of the mold.
Using the wire, the mold is immersed (for 10 seconds) in a beaker of
isopentane chilled in liquid nitrogen. The tissue is stored in a -70
freezer. When ready to cut, a thin layer of OCT is placed on a cryostat
chuck, the tissue is removed from the foil and placed upside down on the
chuck. It is flat, the orientation is maintained, we have seen no freeze
artifact with the biopsies. 10 sections are cut at 2 microns, 1 section
per slide. Dried for 30 minutes then stained.

Rena Fail
Medical University of SC
Charleston, SC

-----Original Message-----
From: Lisa Black [] 
Sent: Wednesday, July 16, 2003 2:53 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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