FLIM-FRET in solutions: In vitro measurement of a Cerulean-Citrine biosensor

For imaging purposes, FRET-based probes have been designed to study protein interactions and conformational changes of molecules like protease sensors. FRET efficiency can be measured based on fluorescence lifetime microscopy (FLIM). FLIM-FRET allows analysis of molecular interactions both in vitro and in vivo. This article describes the use of FLIM in the time domain (TCSPC) to measure FRET in vitro in a biochemical assay using a Cerulean-Citrine construct.

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