Hi Amy:

Amy Janes wrote:

>Hi,
>I wrote before because my mouse brain tissue was full of holes and I was
>told I should flash freeze the tissue, which I did and it seemed to work.
>However, when I used this technique I cut the tissue immediatley after
>freezing. Now I am running ICC on tissue that was flash frozen and then
>stored at -20C. 
>
How long was the storage? Did you store sections or the tissue block? 
Minus 20 may not be cold enough to retain antigenicity, some antigens 
degrade at minus 80.

>The tissue again looks like it has tiny holes. I am
>running ICC for a retrograde tracer which is supposed to label cell bodies but instead looks like a smear of color and I cannot see individual cells.
>
Is this fixed or fresh frozen tissue?

>I am double labeling for fos and I get no staining (because I think the
>cells are destroyed). Any suggestions? 
>
Have you done each label by itself to see the results? If so, how do 
they look? How does your known positive control look?

>Is there a better/safer way to
>store tissue if I won't be able to cut it quickly?
>
Depends on the antigen and how you are storing the tissue. First off, 
-20 may not be cold enough but I need to know more (see above).

Geoff

>
>Thanks,
>
>Amy Janes, MA
>Boston University
>Department of Psychology
>Molecular Neurobiology
>and Behavior Laboratory
>
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>  
>

-- 
--
**********************************************
Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff@umdnj.edu
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