Kniss Ariel, Lu Hang, Jones Dean P, Kemp Melissa L
The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
Methods Enzymol. 2013;526:219-30. doi: 10.1016/B978-0-12-405883-5.00013-2.
Most current studies of reactive oxygen species (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.
目前大多数关于活性氧（ROS）产生的研究报告的是细胞内的全局平均测量值；然而，ROS可在不同的亚细胞位置产生，并在其紧邻区域产生局部效应。一个用于高通量单细胞成像的微流控平台能够监测线粒体ROS产生在空间和时间上的变化。通过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的贡献者——线粒体过氧化氢或超氧化物进行单细胞监测。用抗霉素A刺激Jurkat T淋巴瘤细胞，并随时间在阵列微流控装置中成像。观察到单细胞反应的差异是抑制剂浓度和所使用的ROS测量类型的函数。
