[Histonet] Mouse Autoradiography of fixed frozen tissue

[Sebree Linda A.]

Its been a long time Jamie, but when I did autoradiography on frozen
rabbit heart, I simply cut the frozen sections in a dark room, air-dried
and dipped in emulsion.  Then I sealed them in a light-tight box to
develop.  

Linda A. Sebree
University of Wisconsin Hospital & Clinics
IHC/ISH Clinical & Research Laboratory
DM223-VA
600 Highland Ave.
Madison, WI 53792
(608)265-6596
FAX:  (608)262-7174



-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jamie E
Erickson
Sent: Friday, December 16, 2005 1:20 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue


Hello everyone,
                          I was hoping that I can get some assistance on
a 
project I'm going to do. An Investigator here would like to look at 
radiolabeled mouse Sm. intestines for there specific radiolabeled
protein. 
My problem is that I don't want to use our only processor to process 
radioactive material ( I 125 or H3, not sure yet) My other histology 
colleges may not like me so much if I did that.
       So I plan on fixing the material in 10% NBF (24-48 hours),
washing 
in water (Blot)  then cryopreserving them (LN2 Isopentane). This way I
can 
Cryosection and decontaminate the less used cryostat.  Next I  wash in 
water and lastly dipping slides in Photographic Emulsion and wait, and 
wait....My question is, has anyone done this that could help me out with

the finer details and will this be OK?  I've done this with paraffin 
samples dip wait 2,4,7,14, 21, etc days for a signal, but not with 
frozens.. 

If anyone has done this and could help me out I'd really appreciate it..

Thanks

Jamie

_______________________________
Jamie Erickson
Sr. Research Associate 
Department: DSMP
Abbott Bioresearch Center
100 Research Drive
Worcester, MA 01605-4341
508-688-3134
FAX: 508-793-4895
e-mail: jamie.erickson <@t> abbott.com
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