Rare earth cryptates for the investigation of molecular interactions in vitro and in living cells

摘要: We previously developed a technique known as homogeneous time-resolved fluorescence (HTRF) which has become reference method in the design of bioassays. The makes use resonance energy transfer (FRET) depending close proximity between europium cryptate, long-lived luminescent donor, and matching fluorescent acceptor. extension this to vivo cellular imaging is possible by labeling specific molecules within living cell, each labeled either with donor or acceptor taking images (FRET). mode allows discriminate signal arising from background excess amounts unbound marker cell autofluorescence. When combination having longer lifetime an shorter used, measured decays acceptors under FRET becomes slower than decay upon direct excitation. also much autofluorescence, interference autofluorescence reduced microscope time-gated function equipped camera. was evaluated both microplate TRF reader.