[Histonet] microglia

[Antonia Abeyta]

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.

Thanks,
Antonia

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