FRET
Förster Resonance Energy Transfer (FRET) is the transfer of photonic energy between two light-sensitive molecules (chromophores). When both chromophores are fluorophores, this is often referred to as Fluorescent Resonance Energy Transfer. The ‘donor’ molecule moves to an excited state after absorbing a photon. The ‘donor’ can then transfer this energy to the ‘acceptor’ chromophore via nonradiative dipole-dipole coupling. This energy transfer can only happen if the donor and acceptor are in close proximity to one another, making this technique very sensitive to small changes in chromophore separation. To image FRET effectively, one must clearly quantify both the donor and the acceptor emission to ensure that any FRET signal does not contain direct fluorescence from either donor or acceptor chromophores.
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