Quantitative analysis of fluorescence lifetime imaging made easy
Quantitative analysis of fluorescence lifetime imaging made easy
复制标题
荧光寿命成像的定量分析变得简单
作者：
F. Wouters;A. Esposito
荧光寿命成像是研究活细胞中荧光团分子环境的一种有价值的、通用的工具。它非常适合-因此越来越多地用于定量Förster共振能量转移的发生，这是一种用于检测亚纳米构象变化，蛋白质-蛋白质相互作用和蛋白质生化状态的强大显微镜方法。然而，仔细的定量分析是需要正确和有意义的解释荧光寿命数据。对于非专业用户来说，这可能是一项艰巨的任务，并且是许多可避免的错误和不合理解释的来源。Digman及其同事（Digman et al., 2007，《生物物理学》）。J. 94, L14-6)提出了一种分析技术，该技术避免了数据拟合，而倾向于简单的图形极坐标数据表示。在这个“相量”空间中，对于没有经验的用户来说，终身成像的物理学变得更加直观和容易理解。从图像像素累积的信息，甚至在不同的细胞上，描述了可以用物理上有意义的方式在视觉上解释的模式和轨迹。在uPAR受体二聚化的研究中证明了它的有用性。
Fluorescence lifetime imaging is a valuable and versatile tool for the investigation of the molecular environment of fluorophores in living cells. It is ideally suited— and is therefore increasingly used—for the quantification of the occurrence of Förster Resonance Energy Transfer, a powerful microscopy method for the detection of subnanometer conformational changes, protein‐protein interactions, and protein biochemical status. However, careful quantitative analysis is required for the correct and meaningful interpretation of fluorescence lifetime data. This can be a daunting task to the nonexpert user, and is the source for many avoidable errors and unsound interpretations. Digman and colleagues (Digman et al., 2007, Biophys. J. 94, L14–6) present an analysis technique that avoids data fitting in favor of a simple graphical polar data representation. In this “phasor” space, the physics of lifetime imaging becomes more intuitive and accessible also to the inexperienced user. The cumulated information from image pixels, even over different cells, describes patterns and trajectories that can be visually interpreted in physically meaningful ways. Its usefulness is demonstrated in the study of the dimerization of the uPAR receptor.
影响因子：
3.4
作者：
Digman, Michelle A.;Gratton, Enrico
通讯作者：
Gratton, Enrico
影响因子：
3.4
作者：
Digman, MA;Brown, CM;Gratton, E
通讯作者：
Gratton, E