Our EM Tech here has a very clever method of doing 
ultramicrotomy of tissue from a microtome or cryostat section. The 
thick sections are observed with a light mic, and the area of interest 
is selected. She then places a drop of epon on the area of interest, 
removes the solidified drop, and ultramicrotomizes (sounds almost 
dirty doesn't it?) the section. IEM will then be done on the thins.
Cheers! Greg

Date sent:      	Sun, 09 Jul 2000 00:46:01 -0300
From:           	Gayle Callis <uvsgc@msu.oscs.montana.edu>
Subject:        	Re: Revisiting storage of cryosections
Forwarded to:   	DOBBIN@acad1.cs.upei.ca
To:             	Greg Dobbin <dobbin@upei.ca>

> I am not sure what you mean by doing immunoelectron microscopy on a frozen
> section, a special ultra-cryomicrotomy technic???
> 
> Frozen sections should be fixed and stored at -80C, in an air and light
> tight box, with a bag of silica gel to maintain dry conditions.  Bring to
> RT for 30 min with lid on, to avoid condensation on section.  
> 
> Sometimes antigens survive being frozen unfixed, but may or may not work.
> You can take a fixed section, rinse in PBS to remove all aldehyde
> fixatives, and rinse in PBS with 1% BSA for 10 in and store.  This protein
> will protect the section. 
> 
> Our frozen sections are cut at 5 um in a cryostat, for light microscopy,
> not EM.
> 
>  
> 
> At 11:37 AM 7/19/00 -0300, you wrote:
> >Hello Histonetters,
> >I do recall this subject being discussed before, but at the time I 
> >read with passing interest. I now have a graduate student 
> >wondering how long an air-dried frozen section can be stored and 
> >under what conditions, prior to doing immunoelectron microscopy.
> >He would like to batch process his sections. Any suggestions?
> >Cheers! Greg
> >>
> >>
> >>
> >>
> >>
> >>
> >Greg Dobbin
> >Pathology Lab
> >Atlantic Veterinary College, U.P.E.I.
> >550 Unviversity Ave.
> >Charlottetown, P.E.I.
> >Canada,  C1A 4P3
> >Phone: (902)566-0744
> >Fax: (902)566-0851
> >
> >
> >
> Gayle Callis
> Veterinary Molecular Biology
> Montana State University
> Bozeman MT 59717-3610
> 406 994-4705
> 406 994-4303


>
>
>
>
>
>
Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 Unviversity Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851