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A microfluidic systems biology approach for live single-cell mitochondrial ROS imaging

机译:活单细胞线粒体ROs成像的微流体系统生物学方法

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摘要

Most current studies of ROS production report globally averaged measurements within the cell; however, ROS can be produced in distinct subcellular locations and have local effects in their immediate vicinity. A microfluidic platform for high-throughput single cell imaging allows mitochondrial ROS production to be monitored as varying in both space and time. Using this systems biology approach, single cell variability can be viewed within a population. We discuss single cell monitoring of contributors to mitochondrial redox state - mitochondrial hydrogen peroxide or superoxide - through the use of a small molecule probe or targeted fluorescent reporter protein. Jurkat T lymphoma cells were stimulated with antimycin A and imaged in an arrayed microfluidic device over time. Differences in single cell responses were observed as a function of both inhibitor concentration and type of ROS measurement used.
机译:最新的ROS产生研究报告了细胞内的全球平均测量值。然而,ROS可以在不同的亚细胞位置产生,并在其附近具有局部作用。用于高通量单细胞成像的微流控平台允许监测线粒体ROS的产生,因为其时空变化。使用这种系统生物学方法,可以在群体中观察单细胞变异性。我们讨论了通过使用小分子探针或靶向荧光报告蛋白,对线粒体氧化还原状态(线粒体过氧化氢或超氧化物)的贡献者进行单细胞监测。用抗霉素A刺激Jurkat T淋巴瘤细胞,并随时间在阵列微流控设备中成像。观察到单细胞反应的差异是抑制剂浓度和所用ROS测定类型的函数。

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