[Histonet] fresh-frozen mouse tail sectioning

[Patsy Ruegg]

I would just lay it on it's side for freezing the first time and then turn
it vertically to mount onto a chuck for sectioning.  Also, you probably do
not have this, but for difficult things like this (the tail will have at
least cartilage in it) I use the tape transfer technique from Instrumedics
(now owned by McCormick).
Patsy

Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 215
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Gaupp, Dina
D 
Sent: Friday, May 16, 2008 10:05 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] fresh-frozen mouse tail sectioning

To Whomever This Applies:
 
I am having big problems sectioning mouse tail, approximately 2mm in
thickness, vertical embedded in OCT for frozen sections.  The tissue is
fresh-frozen because the principle investigator would like to detect an
enzyme x-gal.  No fix, no cryopreservation - the tissue was not immersed in
any type of solution.  He snipped the ends of a mouse tail & immediately
gave me the tissues.  I embedded the tissue vertically in OCT & flash froze
at -80C.  Upon sectioning, the tissue rolled.  I could not for the life of
me get 1 section.  All the tissues rolled & its so tiny to begin with that
it was hard for me to grab it.  I used the anti-roll plate, hoping it would
hold the tissue in place but it still rolled.  I changed temperature
settings(increase & decrease temp), thickness, angle, rubbed it with my
fingers.  Everything I could think of to get a section.  I asked someone
else in the lab to try & they couldn't get a section.  It was like the
tissue completely separated from OCT.  The principle investigator will give
me more samples & I don't want this to happen again.  
 
Can anyone in histoland help me, tell me what I didn't do or what I did
wrong?
 
Dina D. Gaupp, BS, MT
Senior Lab Supervisor
Center for Gene Therapy, SL-99
Tulane University Health Science Center
1430 Tulane Ave
New Orleans, La 70112
Lab: 504-988-1194
dgaupp <@t> tulane.edu <mailto:dgaupp <@t> tulane.edu> 
 
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