Assessment of mitochondrial membrane potential using an on-chip microelectrode in a microfluidic device
Assessment of mitochondrial membrane potential using an on-chip microelectrode in a microfluidic device
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DOI：
10.1039/c001818j
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Burke, Peter
中科院分区：
文献类型：
--
作者：
Lim, Tae-Sun;Davila, Antonio;Burke, Peter
线粒体膜电位用于产生和调节生命系统中的能量，驱动 ADP 转化为 ATP，调节离子稳态并控制细胞凋亡，所有这些对于人类健康和疾病都至关重要。因此，需要一种工具来研究其在受控环境中的调节，以实现潜在的临床和科学应用。为此，在微流体环境中构建了片上四苯基磷 (TPP+) 选择性微电极传感器。膜电位测量中使用的分离线粒体 (Heb7A) 浓度为 0.3 ng mu L-1，比传统测定中使用的浓度 (3 μ g mu L-1) 小四个数量级。此外，腔室体积（85μL）比传统实验小2个数量级。作为演示，可以清楚地测量膜电位的变化，以响应电子传递链的一系列众所周知的底物和抑制剂。这种通用方法迄今为止尚未在线粒体功能和生物能量学研究中得到证实，但它可以通过在受控（微流体）化学环境中对线粒体膜电位响应细胞凋亡抑制剂和诱导剂的调节、动力学和统计特性进行高通量研究，有助于推进线粒体研究和临床应用领域。
The mitochondrial membrane potential is used to generate and regulate energy in living systems, driving the conversion of ADP to ATP, regulating ion homeostasis, and controlling apoptosis, all central to human health and disease. Therefore, there is a need for tools to study its regulation in a controlled environment for potential clinical and scientific applications. For this aim, an on-chip tetraphenylphosphonium (TPP+) selective microelectrode sensor was constructed in a microfluidic environment. The concentration of isolated mitochondria (Heb7A) used in a membrane potential measurement was 0.3 ng mu L-1, four orders of magnitude smaller than the concentration used in conventional assays (3 mu g mu L-1). In addition, the volume of the chamber (85 mu L) is 2 orders of magnitude smaller than traditional experiments. As a demonstration, changes in the membrane potential are clearly measured in response to a barrage of well-known substrates and inhibitors of the electron transport chain. This general approach, which to date has not been demonstrated for study of mitochondrial function and bio-energetics in generally, can be instrumental in advancing the field of mitochondrial research and clinical applications by allowing high throughput studies of the regulation, dynamics, and statistical properties of the mitochondrial membrane potential in response to inhibitors and inducers of apoptosis in a controlled (microfluidic) chemical environment.