荧光共振能量转移-荧光相关光谱技术：推动对复杂生物系统的全面洞察。

FRET-FCS: Advancing comprehensive insights into complex biological systems.

作者信息

Lazaro-Alfaro Anay, Nicholas Sterling L N, Sanabria Hugo

机构信息

Department of Physics and Astronomy, Clemson University, Clemson, South Carolina.

出版信息

Biophys J. 2025 Apr 15. doi: 10.1016/j.bpj.2025.04.015.

DOI:10.1016/j.bpj.2025.04.015

PMID:40241306

Abstract

Förster resonance energy transfer (FRET) is a short-range distance-dependent photophysical phenomenon that allows the measurement of intra- and intermolecular distances through fluorescence detection. FRET measurements are sensitive to the movements of fluorescently labeled molecules as they produce fluorescence fluctuations. Fluorescence correlation spectroscopy (FCS) analyzes these fluctuations at faster and broader timescales (from picoseconds to seconds) compared with other techniques, unraveling the thermodynamic and kinetic properties of the system under study. Therefore, the combination of FRET and FCS (FRET-FCS) facilitates the analysis of molecular dynamics. Since its introduction, FRET-FCS has evolved into studying more sophisticated systems, requiring improvements in data acquisition and analysis. In this review, we discuss applications in the field of FRET-FCS that propose novel alternatives to overcome the inherent limitations of experimental setups. This work aims to promote using and enhancing FRET-FCS techniques to develop a comprehensive understanding of biological systems.

摘要

Förster共振能量转移（FRET）是一种短程距离依赖性光物理现象，它允许通过荧光检测来测量分子内和分子间的距离。FRET测量对荧光标记分子的运动敏感，因为它们会产生荧光波动。与其他技术相比，荧光相关光谱（FCS）能在更快、更宽的时间尺度（从皮秒到秒）上分析这些波动，从而揭示所研究系统的热力学和动力学性质。因此，FRET与FCS的结合（FRET-FCS）有助于分子动力学分析。自引入以来，FRET-FCS已发展到研究更复杂的系统，这需要改进数据采集和分析。在本综述中，我们讨论了FRET-FCS领域的应用，这些应用提出了新颖的替代方法来克服实验装置的固有局限性。这项工作旨在促进FRET-FCS技术的使用和改进，以全面了解生物系统。