Quantitative In Vivo Imaging of Molecular Distances Using FLIM-FRET
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Förster Resonance Energy Transfer (FRET) is a valuable tool for measuring distances on the nanometer scale. In this process, energy from an excited donor molecule is transferred to a nearby non-excited acceptor. As the energy transfer process is strongly distance dependent, the quenching of donor fluorescence can be directly correlated to the donor-acceptor distance. The article presented here provides a description of using FRET in combination with Fluorescence Lifetime imaging (FLIM) to map out molecular interaction between labeled fluorescent proteins.
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