RE: [Histonet] Mouse Autoradiography of fixed frozen tissue

From:"Bugelski, Peter [CNTUS]"

Why not process to paraffin "by hand"?  Process through xylene at room temp
and then transfer to paraffin in an oven overnight.  Do everything in
scintillation vials.  This way you can keep the volumes low and check for
leaching radioactivity by either gamma or scintillation counting.  I've done
this many times.

There should be no difference in the exposure time required.

The only steps that need to be in the dark are the melting of the emulsion,
the dipping, the exposur, the development and photographic fixing.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jamie E
Erickson
Sent: Friday, December 16, 2005 2:20 PM
To: histonet@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@abbott.com
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