I am designing a FRET based biosensor. I am wondering whether two fluorscent dyes should physically contact for happening FRET.
I think FRET is caused by the overlap of excitation range of acceptor and emission range of donor. Then, is it possible FRET happen when two dyes are apart between air? (Of course they are apart within very short distance)
I agree with Peter. Atleast find a figure of distance dependence of FRET efficiency, look at that and realize how FRET efficiency changes with distance.
If the two fluorophores (the acceptor need not be fluorescent, by the way) are in physical contact, then the most likely outcome is static quenching, not FRET, which is a short-distance, through-space effect of a dipole-dipole interaction between the fluorophores.
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