Abstract
Fluorescence lifetime is the mean time elapsed between the activation of fluorophore and emission of a photon from the fluorophore. Because the fluorescence lifetime is sensitive to the local environment of the fluorophore and fluorescence resonance energy transfer (FRET), it has been used to monitor intracellular biochemical reaction. In particular, fluorescence lifetime imaging (FLIM) combined with FRET biosensors allows us to monitor the spatiotemporal dynamics of activity of proteins with the resolution of single synapses. In this chapter, we discuss the principle and application of fluorescence lifetime imaging with particular focus on imaging protein activity in single dendritic spines.
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