Förster Resonance Energy Transfer (FRET) is a unique biophysical phenomenon that allows for energy transfer to occur between two light-sensitive molecules, e.g., a donor and an acceptor fluorophore. FRET has been leveraged in numerous biomedical applications to monitor molecular interactions at the nanoscale. Previously, we have developed a FRET-based nanometer-range proximity assay (2–10 nm) that measures receptor-ligand protein complexes during internalization and subsequent endocytic trafficking steps in vitro and in vivo. Recently, there has been great interest to perform FRET imaging in vivo, especially in the context of quantifying drug delivery in vivo.
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