In the near future I would like to be able to detect virally induced
apoptosis in epithelial cells (and eventually in leukocytes) by fixing the
cells and staining with acridine orange for DNA in apoptotic bodies.
However, I also want to look for viral antigen in the same cells by
indirect immunofluorescence.
I know that acridine orange will fluoresce green at 525nm when bound to
DNA (which rules out FITC as a conjugate for my secondary in the viral
antigen IF), but I have also read that acridine orange fluoresces red at
650nm when bound to RNA. I expect to find the viral Ag's in the
cytoplasm... which is where the RNA will be.
The question is which would be the best conjugate to use for the secondary
Ab in light of the fact that the cytoplasm will probably stain red? Is
there a potential conjugate which would fluoresce red at 525nm? Am I
overlooking something fundamental (this is my first foray into IF
staining)?!
Many thanks in advance.
Graeme
--
Graeme Price
Microbial Molecular Genetics and Cell Biology Group,
School of Biological Sciences, Biology West Building,
University of Birmingham,
Edgbaston, Birmingham,
West Midlands, B15 2TT.
United Kingdom.
Tel. (+44) (0)121 414 6555
Fax. (+44) (0)121 414 6557
E-mail g.e.price at bham.ac.uk
