I completely agree with Rik's comment - FRET is a powerful tool to messure
molecular distances beyond the resolution of optical microscopes. However,
this applies only when using the correct filter sets. I was wondering
whether with conventional filter combinations - as generally and very often
used in fluorescence microscopy to detect FITC and rhodamin signals (for
example) - quenching may produce a significant problem for detection of the
donor. Is it possible, that with such classical setups, the truely
colocalizing antigens show up as acceptor fluorescence only, and
consequently, that with dye pairs lacking spectral overlap (as nessessary
for FRET) one might get different double labeling results?
Jacques
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