Fluorescence lifetime imaging (Flim): Basic concepts and recent applications

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.