Assessing mitochondrial potential, calcium, and redox state in isolated mammalian cells using confocal microscopy.
Assessing mitochondrial potential, calcium, and redox state in isolated mammalian cells using confocal microscopy.
复制标题
DOI：
10.1007/978-1-59745-365-3_30
复制
发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
Duchen, Michael R
中科院分区：
文献类型：
--
作者：
Davidson, Sean M;Yellon, Derek;Duchen, Michael R
线粒体在调节细胞内钙动力学中起着至关重要的作用。荧光染料可用于提供氧化还原状态、线粒体膜电位和线粒体钙含量的直接测量。这种方法的简单性使其适合于高通量测定和时间分辨分析;然而，必须小心避免人为结果。我们概述了一般的方法，使用共聚焦显微镜分析的氧化还原状态，线粒体膜电位，和线粒体钙含量在成年心肌细胞。我们演示了如何使用发射光谱的“指纹”方法，即使发射光谱重叠，这些参数可以并行分析。
Mitochondria play a vital role in the regulation of intracellular calcium dynamics. Fluorescent dyes can be used to provide a direct measurement of the redox state, mitochondrial membrane potential, and mitochondrial calcium content. The simplicity of this approach lends itself to high-throughput assays and time-resolved analyses; however, care must be taken to avoid artifactual results. We outline general methods using confocal microscopy for analysis of the redox state, mitochondrial membrane potential, and mitochondrial calcium content in adult cardiomyocytes. We demonstrate how these parameters can be analyzed in parallel using the emission spectra "fingerprinting" method even when emission spectra overlap.