荧光共振能量转移-荧光寿命显微成像(FRET-FLIM)技术在生命科学研究中的应用进展

Advances in the Application of F rster Resonance Energy Transfer andFluorescence Lifetime Imaging Microscopy (FRET-FLIM)Technique in Life Science Research

细胞是动植物结构和生命活动的基本单位。细胞过程的一个重要特点就是其生化组分在时空调控上的相互作用关系。然而,利用传统的生化方法(如酵母双杂交系统、pull-down系统等)很难在空间上评估活细胞内分子间的相互作用。光学技术的快速发展,为研究活细胞中生物分子的时空动态提供了新的遗传研究工具,其中荧光共振能量转移-荧光寿命显微成像(FRET-FLIM)技术在实时探测分析活细胞中生物大分子构象变化和分子间动态相互作用过程具有独特的优势,如:实现对活细胞的实时“可视化”研究,同时具有高时空分辨率;检测更加灵敏、结果可信度高;且基于简易的数学运算完成简单快捷的分析程序。介绍FRET-FLIM技术的理论背景知识,对比了该技术与传统蛋白相互作用技术研究的利弊,同时归纳了其在蛋白相互作用、细胞生物学和疾病诊断等方面的最新应用研究进展,最后总结和讨论了FRET-FLIM技术的未来发展趋势,以期能够为揭示活细胞的结构和细胞过程相关研究提供新的见解。

Cell is the basic unit of structure and biological activities of animals and plants.An important feature of cellular processes is that the biochemical components involved in it have spatiotemporally interacted with each other.However,it is difficult to evaluate molecular interactions in living cells using traditional biochemical methods(e.g.,yeast two-hybrid system,pull-down system,etc.).The rapid development of optical technology provides new genetic research tools for studying the spatiotemporal dynamics of biomolecules in living cells.FRET-FLIM(F rster resonance energy transfer-fluorescence lifetime imaging microscopy)technology has unique advantages in real-time detection and analysis of conformational changes and dynamic interactions of macromolecules in living cells,such as achieving real-time“visualization”study of living cells with the high spatial and temporal resolution,obtaining more reliable results with high sensitivity,and accomplishing simple and fast analysis program based on brief mathematical operations etc.This paper outlined the theoretical backgrounds of FRET-FLIM,and then summarized the advantages and disadvantages of FRET-FLIM technique compared with traditional methods for protein interaction studies.In addition,we emphasized the recent studies and applications of FRET-FLIM in protein interaction,cell biology,and disease diagnosis.Finally,this paper prospected the future research trend of FRET-FLIM in the field of life sciences and provided a comprehensive and systematic reference for related research in the structure and processes of living cells.