Quenching FITC

RICHARD P. PHIPPS PHIP at BPHVAX.BIOPHYSICS.ROCHESTER.EDU
Thu Jun 16 09:17:07 EST 1994


In <2tnt72$hc8 at news.duke.edu> mjtino at acpub.duke.edu writes:

> I am having trouble with an assay for phagocytosis in which I would like to 
> examine fluorescence of ingested FITC-labelled bacteria by macrophages.
> In order to do this, I must quench the FITC outside of the macrophages to
> distinguish between extra- and intracellular fluorecence.  I've
> tried crystal violet and trypan blue, but with no success.  Any suggestions?
> 
> Michael Tino
> michael_tino at cellbio.duke.edu
> 
> 

I seem to remember a protocol where the cells you wish to assay 
(macrophages in this case) are labelled red (don't remember whether
they used PE or EtBr or something like that).  Under a fl. microscope,
extracellular bacteria is green and phagocytosed bacteria is sort of
yellow-orange.  If I remember the name of the author I will repost.

Rick Willis
University of Rochester Cancer Center




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