[Histonet] Problems with cutting frozen sections of skin punc h biopsies

Dawson, Glen GDawson <@t> dynacaremilwaukee.com
Mon Dec 5 07:48:15 CST 2005


Place your outside skin punch into a beaker (I use a 100 ml. beaker) filled
3/4 full with something like a pH 7.6 Tris buffer (if you have no buffer, DI
water will work better than nothing).  Put your beaker on a stirring plate &
add a stir bar to the buffer that now contains the bx.  Stir (I stir at
about 50% speed) for ten minutes.  If the bx. is especially large repeat the
previous steps again with fresh Tris.  

Get your cryostat chuck ready, place your bx on a paper towel to soak off
the excess buffer, mount & cut.  Unless you rinse out the Michel's transport
medium, you will mainly get an empty hole where the tissue was.  You'll be
amazed at how much better it will cut after a good rinse.

Good Luck,

Glen Dawson  BS, HT & QIHC (ASCP)
IHC Manager
Milwaukee, WI

-----Original Message-----
From: nina.owen <@t> tiscali.co.uk [mailto:nina.owen <@t> tiscali.co.uk]
Sent: Saturday, December 03, 2005 2:18 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Problems with cutting frozen sections of skin punch

Hi all!

Just wondered if anyone could help us with the following problem...

Our histology lab occasionally receives fresh skin punch biopsies for direct
immunofluorescence from the hospital's dermatology department.  When
frozen sections from these specimens we don't experience any problems during
cutting them.  However, we are increasingly receiving punch biospies from
other hospitals for DIF staining; these are sent in Michael's medium for
preservation on transit.  We seem to have a lot of difficulty cutting these!
 The problem is that the sections seem to crease up immediately, or if we
get a decent section the tissue ends up sticking to the anti-roll plate.
 Thus, when the anti-roll plate is lifted to allow the section to be picked
up onto a slide, the tissue is lifted out of the OCT mountant (Shandon)
a hole!

We have tried a couple of things to try and stop this from happening.  If
we manage to get the specimen into the deep freeze asap and leave it for
a few days it is slightly better.  Also, leaving the tissue mounted on a
chuck in the cryostat to cool down for about an hour prior to attempting
to cut it stops the tissue from sticking to the anti roll plate, but only
long enough for about two sections to be obtained.

Has anyoneelse had this problem with fresh specimens that have been in
medium?  If so, how did you sort it?

Thanks Histonetters!

Nina Owen
Dewsbury and District Hospital, Mid Yorkshire NHS Trust


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