Re: [Histonet] microglia

From:Geoff McAuliffe

Hi Antonia:

    What animal and what antibody are you using? Mac1 or F4/80 work fine 
on frozen sections of  mouse CNS. For rat CNS, OX-42 works on frozen 
sections. Lots of literature on these. Can you get an animal to fix by 
perfusion so you could compare your results on saline-fresh frozen 
versus perfused?


Antonia Abeyta wrote:

>Hi all,
>Has anyone ever stained for microglia on frozen tissue?  Our brain
>tissue is perfused with saline and I have tried fixing with 4%
>paraformaldehyde and PLP.  My results for both were similar, I am seeing
>some microglia, but the morphology is not like it should be, and
>contrast isn't very good.  I'm also having to use a very concentrated
>diltution (1:250) of an antibody that has worked at 1:1000 for some of
>my collegues.  The difference is they are perfusing with PLP and using
>paraffin rather than frozen sections.  Since we are using the sections
>for other experiments we are stuck perfusing w/saline and frozen
>sections...I'm about out of ideas and could use any suggestions!  I am
>trying to get this to work for both fluorescence and ABC/DAB.
>Antonia L. Abeyta
>Health Sciences Tech. III
>Community Environmental Health Program
>University of New Mexico
>Surge Bldg. Room 140
>Albuquerque, NM  87131
>(505) 272-4028
>Histonet mailing list

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

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