Hi Phoebe,
	If the researcher would like a technique that I have used
successfully for 30 years, he/she may contact me as below.  Most of my work
has been with 3H, but depending on the nuclide some must be dealt with
differently, at least with greater care.
	Few tissue specimens will be 'hot' enough to cause you harm, but you
must deal with safety for each nuclide differently as well.  32P must be
handled not only carefully but quickly.  As an example, 125I will
concentrate in the thyroid rather than uniformly throughout the body.
3H-Me-Thymidine will disappear from the circulation within about 30min, and
any that is metabolized, i.e. to Uridine, will lose its Me which is the
'hot' part.  Labeling of animals and tissues is generally performed using
radioactive labels whose doses are either 1uCi/g body weight or 1uCi/ml of
culture fluid.  There are often chases with non-radioactive label following
the labeling dose.  Any dose of tritium or 14C that exceeds 1uCi/g or
1uCi/ml will likely cause rapid radiation damage in the experimental
specimen, so you can see that limits are followed.  I always have used
0.5uCi/g!
	Note:  labels that remain soluble must be treated with VERY special
methods to prevent loss and/or diffusion, but usually one doesn't care what
happens to un-incorporated label.

Here's a good site for In Situ + Autoradiography

My protocol is in word/PDF

Regards,

Fred Monson

Frederick C. Monson, PhD   
Center for Advanced Scientific Imaging
Schmucker II Science Center
West Chester University
South Church Street and Rosedale
West Chester, Pennsylvania, USA, 19383
Phone:  610-738-0437
FAX:  610-738-0437
fmonson@wcupa.edu
CASI URL:  http://darwin.wcupa.edu/casi/
WCUPA URL:  http://www.wcupa.edu/
Visitors URL:  http://www.wcupa.edu/_visitors/


> ----------
> From: 	Phoebe J Doss/app/Cvm
> Sent: 	Wednesday, May 1, 2002 5:22 PM
> To: 	microscopy@sparc5.microscopy.com
> Subject: 	sectioning "hot" tissue
> 
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> 
>  
> 
>                       "Ginger
> 
>                       Hendricks"               To:       "Doss, Phoebe and
> Mike"   
>                        Doss/app/Cvm)                  
>                       net>                     Subject:  please forward to
> the MSA listserv            
>  
> 
>                       05/01/02 04:13 PM
> 
>  
> 
>  
> 
> 
> 
> 
> 
> Hello all,
> 
> I have a researcher who is interested in using autoradiography with nerve
> tissue.  His tech is researching methods and has come to me for advice.  I
> have never performed this procedure and am concerned about sectioning
> "hot"
> (S35) tissue with the microtome.  Any protocols, suggestions or direction
> would be greatly appreciated.
> 
> My email address is grhendricks@cox.net
> 
> Thank you in advance,
> 
> Ginger
> 
>                             Ginger R. Hendricks
>                    EM Lab Manager and Adjunct Instructor
>                                   OSU-CHS
>                              1111 W. 17th St.
>                               Tulsa, OK 74107
>                             (918) 561-8232 work
>                             (918) 699-8629 fax
>                             grhendricks@cox.net
>                                  Website:
> http://www.healthsciences.okstate.edu/research/rsp/electronmicroscopy/elec
> tron_microscopy_laboratory.htm
> 
> 
> 
> 
> 
>