Solaini Giancarlo, Sgarbi Gianluca, Lenaz Giorgio, Baracca Alessandra
Dipartimento di Biochimica, Università di Bologna, Via Irnerio 48, Bologna, 40126, Italy.
Biosci Rep. 2007 Jun;27(1-3):11-21. doi: 10.1007/s10540-007-9033-4.
Permeant cationic fluorescent probes are widely employed to monitor mitochondrial transmembrane potential and its changes. The application of such potential-dependent probes in conjunction with both fluorescence microscopy and fluorescence spectroscopy allows the monitoring of mitochondrial membrane potential in individual living cells as well as in large population of cells. These approaches to the analysis of membrane potential is of extremely high value to obtain insights into both the basic energy metabolism and its dysfunction in pathologic cells. However, the use of fluorescent molecules to probe biological phenomena must follow the awareness of some principles of fluorescence emission, quenching, and quantum yield since it is a very sensitive tool, but because of this extremely high sensitivity it is also strongly affected by the environment. In addition, the instruments used to monitor fluorescence and its changes in biological systems have also to be employed with cautions due to technical limits that may affect the signals. We have therefore undertaken to review the most currently used analytical methods, providing a summary of practical tips that should precede data acquisition and subsequent analysis. Furthermore, we discuss the application and feasibility of various techniques and discuss their respective strength and weakness.
渗透性阳离子荧光探针被广泛用于监测线粒体跨膜电位及其变化。这种电位依赖性探针与荧光显微镜和荧光光谱法结合使用，能够监测单个活细胞以及大量细胞中的线粒体膜电位。这些分析膜电位的方法对于深入了解基本能量代谢及其在病理细胞中的功能障碍具有极高的价值。然而，由于荧光分子是一种非常敏感的工具，利用其探测生物现象时必须遵循一些关于荧光发射、猝灭和量子产率的原理，而且正因为这种极高的敏感性，它也极易受到环境的影响。此外，由于可能影响信号的技术限制，用于监测生物系统中荧光及其变化的仪器也必须谨慎使用。因此，我们着手回顾当前最常用的分析方法，总结在数据采集和后续分析之前应遵循的实用技巧。此外，我们还讨论了各种技术的应用和可行性，并探讨了它们各自的优缺点。
