Evaluating mitochondrial membrane potential in cells
Evaluating mitochondrial membrane potential in cells
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DOI：
10.1007/s10540-007-9033-4
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发表时间：
2007-06-01
影响因子：
4
通讯作者：
Baracca, Alessandra
中科院分区：
文献类型：
--
作者：
Solaini, Giancarlo;Sgarbi, Gianluca;Baracca, Alessandra
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。这种电位依赖的探针与荧光显微镜和荧光光谱学相结合，可以监测单个活细胞和大量细胞中的线粒体膜电位。这些膜电位的分析方法对于深入了解病理细胞中的基本能量代谢及其功能障碍具有极高的价值。然而，使用荧光分子探测生物现象必须遵循荧光发射、猝灭和量子产额的一些原理，因为它是一种非常敏感的工具，但由于其极高的灵敏度，它也受到环境的强烈影响。此外，由于可能影响信号的技术限制，用于监测生物系统中的荧光及其变化的仪器也必须谨慎使用。因此，我们承诺审查目前最常用的分析方法，提供在数据获取和后续分析之前应提供的实用技巧的摘要。此外，我们还讨论了各种技术的应用和可行性，并讨论了它们各自的优缺点。
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.