SIMULTANEOUS IMAGING OF CELL AND MITOCHONDRIAL-MEMBRANE POTENTIALS
SIMULTANEOUS IMAGING OF CELL AND MITOCHONDRIAL-MEMBRANE POTENTIALS
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DOI：
10.1016/s0006-3495(89)82754-7
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发表时间：
1989-12-01
影响因子：
3.4
通讯作者：
LOEW, LM
中科院分区：
文献类型：
--
作者：
FARKAS, DL;WEI, MD;LOEW, LM
带电荷的膜渗透性分子探针在细胞内细胞器、细胞质和外部介质之间的分布由这些区域的相对膜电位通过能斯特方程描述的耦合平衡来控制。一系列低膜结合和毒性的高荧光阳离子染料（Escherberg，B.，蒙塔纳，M.- D. Weis，J. P. Wuskell，and L. M.勒夫，1988年。Biophys. J. 53：785-794）允许通过数字成像视频显微镜监测这些平衡。我们采用这种技术的组合，以评估，同时，膜电位的细胞和它们的细胞器在原位。我们描述了这种测量的方法和最佳条件，并将该技术应用于同时遵循，具有良好的时间分辨率，在几个培养细胞系的线粒体和质膜电位。化学试剂（离子载体，解偶联剂，电子传输和能量转移抑制剂）在这些电位中的任一个或两个引起的变化的时间过程是很容易定量的，并根据机械的预期。因此，该方法应适用于更微妙和特异性的，生物诱导的细胞潜在变化的研究。
The distribution of charged membrane-permeable molecular probes between intracellular organelles, the cytoplasm, and the outside medium is governed by the relative membrane electrical potentials of these regions through coupled equilibria described by the Nernst equation. A series of highly fluorescent cationic dyes of low membrane binding and toxicity (Ehrenberg, B., V. Montana, M.-D. Weis, J. P. Wuskell, and L. M. Loew, 1988. Biophys. J. 53:785-794) allows the monitoring of these equilibria through digital imaging video microscopy. We employ this combination of technologies to assess, simultaneously, the membrane potentials of cells and of their organelles in situ. We describe the methodology and optimal conditions for such measurements, and apply the technique to concomitantly follow, with good time resolution, The mitochondrial and plasma membrane potentials in several cultured cell lines. The time course of variations induced by chemical agents (ionophores, uncouplers, electron transport, and energy transfer inhibitors) in either or both these potentials is easily quantitated, and in accordance with mechanistic expectations. The methodology should therefore be appliable to the study of more subtle and sepcific, biologically induced potential changes in cells.