Wallrabe Horst, Periasamy Ammasi
Keck Center for Cellular Imaging, Department of Biology, University of Virginia, Gilmer Hall, Charlottesville, Virginia 22904, USA.
Curr Opin Biotechnol. 2005 Feb;16(1):19-27. doi: 10.1016/j.copbio.2004.12.002.
Förster (or fluorescence) resonance energy transfer (FRET) and fluorescence lifetime imaging (FLIM) have moved center stage and are increasingly forming part of multifaceted imaging approaches. They are complementary methodologies that can be applied to advanced quantitative analyses. The widening application of FRET and FLIM has been driven by the availability of suitable fluorophores, increasingly sophisticated microscopy systems, methodologies to correct spectral bleed-through, and the ease with which FRET can be combined with other techniques. FRET and FLIM have recently found use in several applications: in the analysis of protein-protein interactions with high spatial and temporal specificity (e.g. clustering), in the study of conformational changes, in the analysis of binding sequences, and in applications such as high-throughput screening.
Förster（或荧光）共振能量转移（FRET）和荧光寿命成像（FLIM）已成为核心技术，并越来越多地成为多层面成像方法的一部分。它们是互补的方法，可用于先进的定量分析。合适的荧光团的可用性、日益复杂的显微镜系统、校正光谱渗漏的方法以及FRET与其他技术结合的简便性推动了FRET和FLIM应用的不断扩大。FRET和FLIM最近已用于多种应用：分析具有高空间和时间特异性的蛋白质-蛋白质相互作用（例如聚集）、研究构象变化、分析结合序列以及高通量筛选等应用。
