Quantitative real-time imaging of intracellular FRET biosensor dynamics using rapid multi-beam confocal FLIM.

ABSTRACT: Fluorescence lifetime imaging (FLIM) is a quantitative, intensity-independent microscopical method for measurement of diverse biochemical and physical properties in cell biology. It is a highly effective method for measurements of Förster resonance energy transfer (FRET), and for quantification of protein-protein interactions in cells. Time-domain FLIM-FRET measurements of these dynamic interactions are particularly challenging, since the technique requires excellent photon statistics to derive experimental parameters from the complex decay kinetics often observed from fluorophores in living cells. Here we present a new time-domain multi-confocal FLIM instrument with an array of 64 visible beamlets to achieve parallelised excitation and detection with average excitation powers of ~ 1-2 μW

SUBMITTER: Levitt JA 

PROVIDER: S-EPMC7083966 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature