Abstract
With the development of the new detection methods and the function of fluorescent molecule, researchers hope to further explore the internal mechanisms of organisms, monitor changes in the intracellular microenvironment, and dynamic processes of molecular interactions in cells. Fluorescence resonance energy transfer (FRET) describes the energy transfer process between donor fluorescent molecules and acceptor fluorescent molecules. It is an important means to detect protein–protein interactions and protein conformation changes in cells. Fluorescence lifetime imaging microscopy (FLIM) enables noninvasive measurement of the fluorescence lifetime of fluorescent particles in vivo. The FRET–FLIM technology, which is use FLIM to quantify and analyze FRET, enables real-time monitoring of dynamic changes of proteins in biological cells and analysis of protein interaction mechanisms. The distance between donor and acceptor and their respective fluorescent lifetime, which are of great importance for studying the mechanism of intracellular activity can be obtained by data analysis and algorithm fitting.
Highlights
With the continuous developments in optical technologies, the introduction of °uorescence microscopy, biological images can be obtained at the cellular and subcellular levels
There are some requirements for Fluorescence resonance energy transfer (FRET): ̄rst, the distance between donor and acceptor should be less than 10 nm; second, the emission spectrum of donor overlaps with the excitation spectrum of acceptor; third, there is no overlap between the emission spectrum of donor and acceptor; fourth, there is no overlap between the excitation spectrum of donor and acceptor; ̄fth, the dipole of donor and acceptor should have the same direction
The proportion of donor–receptor interaction can be determined by using Fluorescence-lifetime imaging microscopy (FLIM) to quantify FRET, which is important for biologically relevant research
Summary
With the continuous developments in optical technologies, the introduction of °uorescence microscopy, biological images can be obtained at the cellular and subcellular levels. Fluorescence resonance energy transfer (FRET) is a process via which excited donor °uorescent molecules excite nearby acceptor °uorescent molecules. This is an Open Access article published by World Scientic Publishing Company. FRET–FLIM technology uses the °uorescence lifetime change of a sample to detect the dynamic process of the reaction between the donor and acceptor °uorescent molecules. It can obtain lots of information, such as intracellular protein–protein interactions and conformational changes to proteins in the cells.[1,2,3,4,5,6].
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