[Histonet] autofluorescence with vibratome/PFA fixed embryonic brain sections

From:Gayle Callis


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"


Dear All
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.

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)

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