Abstract
Fluorescence lifetime imaging (FLIM) is a key fluorescence microscopy technique to map the environment and interaction of fluorescent probes. It can report on photo physical events that are difficult or impossible to observe by fluorescence intensity imaging, because FLIM is independent of the local fluorophore concentration and excitation intensity. A FLIM application relevant for biology concerns the identification of FRET to study protein interactions and conformational changes, and FLIM can also be used to image viscosity, temperature, pH, refractive index and ion and oxygen concentrations, all at the cellular or sub-cellular level, as well as autofluorescence. The basic principles and some recent advances in the application of FLIM, FLIM instrumentation and molecular probe development will be discussed.
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