[Histonet] autofluorescence with vibratome/PFA fixed embryonic brain
gcallis <@t> montana.edu
Tue Jan 3 14:50:03 CST 2006
Instead of using a FITC conjugated antibody, try a red fluorophore, i.e.
rhodamine (RRX from Jackson) or Texas Red. You will have nice contrast
with the greenish autofluorescence that tends to be worse with PFA
fixation. Do NOT change the fixation, but simply use a different
color. You could also use an antibody conjugated to Alexa 555 (Molecular
Probes) which is more resistant to photobleaching and extremely bright - it
is the equivalent to rhodamine.
From:"Ali Moussavi Nik"
Happy new year
I am doing Immunohistochemistry on 4% PFA fixed, vibrotome sectioned
embryonic brain. My antibody is FITC conjugated
I have a problem with the background green Autoflorescence of the PFA .
Can anyone propose me other method of fixation for embryonic brain.
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-4303 (FAX)
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