Fluorescence lifetime imaging (FLIM) is a key fluorescence microscopy technique to map the environment and interaction of fluorescent probes. It can report on photophysical events that are difficult or impossible to observe by fluorescence intensity imaging, because FLIM is largely independent of the local fluorophore concentration and excitation intensity. Many FLIM applications relevant for biology concern the identification of Förster resonance energy transfer (FRET) to study protein interactions and conformational changes. In addition, FLIM has been used to image viscosity, temperature, pH, refractive index, and ion and oxygen concentrations, all at the cellular level. The basic principles and recent advances in the application of FLIM, FLIM instrumentation, molecular probe, and FLIM detector development will be discussed.
|Title of host publication||Handbook of Photonics for Biomedical Engineering|
|Publisher||Springer, Dordrecht, Netherlands|
|Number of pages||53|
|Publication status||Published - 29 Mar 2017|