I have run into this in the confocal facility I run. People have
brought samples that they have prepped without first consulting me
and we have all sorts of strange results. PE is not a stain of
choice for fluorescence microscopy. It is very popular with the flow
cytometry folks where its rapid bleaching is not an issue, nor
apparently is its excitation by multiple wavelengths. I would
recommend almost any of the other dyes that excite around 560 in
preference to PE. You will also get a stronger (enhanced) signal if
you use your Cd31 as a primary Ab then tag that with a fluorescent
secondary Ab, although you may have specific reasons for not wanting
to do so.
Try TRITC, Texas Red, Alexa 546, CY3. They will all fluoresce red
and will allow your FITC double stain.
Leona Cohen-Gould, M.S.
Sr. Staff Associate
Director, Electron Microscopy Core Facility
Manager, Optical Microscopy Core Facility
Joan & Sanford I. Weill Medical College
of Cornell University
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