Department of Applied Chemistry, Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura, Saitama 338-8570, Japan.
Molecules. 2018 Nov 14;23(11):2972. doi: 10.3390/molecules23112972.
We review the basic concepts and recent applications of two-dimensional fluorescence lifetime correlation spectroscopy (2D FLCS), which is the extension of fluorescence correlation spectroscopy (FCS) to analyze the correlation of fluorescence lifetime in addition to fluorescence intensity. Fluorescence lifetime is sensitive to the microenvironment and can be a "molecular ruler" when combined with FRET. Utilization of fluorescence lifetime in 2D FLCS thus enables us to quantify the inhomogeneity of the system and the interconversion dynamics among different species with a higher time resolution than other single-molecule techniques. Recent applications of 2D FLCS to various biological systems demonstrate that 2D FLCS is a unique and promising tool to quantitatively analyze the microsecond conformational dynamics of macromolecules at the single-molecule level.
我们回顾了二维荧光寿命相关光谱（2D FLCS）的基本概念和最新应用，它是荧光相关光谱（FCS）的扩展，除了分析荧光强度外，还可以分析荧光寿命的相关性。荧光寿命对微环境敏感，并且与 FRET 结合时可以成为“分子标尺”。因此，在 2D FLCS 中利用荧光寿命可以使我们以比其他单分子技术更高的时间分辨率定量地量化系统的非均一性和不同物种之间的转换动力学。2D FLCS 在各种生物系统中的最新应用表明，2D FLCS 是一种独特而有前途的工具，可在单分子水平上定量分析大分子的微秒构象动力学。
