[Histonet] Problems with cutting frozen sections of skin punchbiopsies

[Katri Tuomala]

Hi Nina,
My understanding is, that Michel's is a transport medium and the specimen 
needs to be washed through certain buffers to remove the salts before you 
are able to get good sections. I don't have the buffer recipe on hand, but 
maybe other histonetters can help you there.

Katri
Katri Tuomala
Hamilton, Ontario, Canada

----- Original Message ----- 
From: <nina.owen <@t> tiscali.co.uk>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Saturday, December 03, 2005 3:18 PM
Subject: [Histonet] Problems with cutting frozen sections of skin 
punchbiopsies


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 
obtaining
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) 
leaving
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 
Michael's
medium?  If so, how did you sort it?

Thanks Histonetters!

Nina Owen
Dewsbury and District Hospital, Mid Yorkshire NHS Trust


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