Electrically evoking and electrochemically resolving quantal release on a microchip.
Electrically evoking and electrochemically resolving quantal release on a microchip.
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DOI：
10.1039/b911763f
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发表时间：
2010-01-07
期刊：
影响因子：
6.1
通讯作者：
Rabbitt RD
中科院分区：
文献类型：
--
作者：
Dittami GM;Rabbitt RD
应用微芯片对大鼠嗜铬细胞瘤(PC12)细胞进行电去极化，同时用安培法检测胞外儿茶酚胺的释放。结果表明，细胞在微通道中通过恒电位电路产生的电场流动时会引起胞吐，细胞外电压脉冲也会触发胞吐。电气有限元模型分析表明，由于较小的分流路径和较高的电流阻力，较大的电池经历了较大的细胞外电场的去极化激发。与这些模拟一致，从细胞团和大细胞记录的数据相对于来自较小细胞的数据表现出更高的释放率。总体而言，该系统能够解析齐托莫尔范围内的单个囊泡量子释放，以及与囊泡融合过程相关的动力学。对尖峰种群统计数据的分析表明，从细胞周围的多个释放部位检测到儿茶酚胺。这种装置在流式细胞术中被用来激发和检测胞吐作用的潜力被证明。
A microchip was applied to electrically depolarize rat pheochromocytoma (PC12) cells and to simultaneously detect exocytotic catecholamine release amperometrically. Results demonstrate exocytosis elicited by flowing cells through an electric field generated by a potentiostat circuit in a microchannel, as well as exocytosis triggered by application of an extracellular voltage pulse across. Electrical finite element model (FEM) analysis illustrated that larger cells experienced greater depolarizing excitation from the extracellular electric fields due to the smaller shunt path and higher resistance to current flow in the channel around the cell. Consistent with these simulations, data recorded from cell clusters and large cells exhibited increased release rates relative to data from the smaller cells. Overall, the system was capable of resolving single vesicle quantal release, in the zeptomole range, as well as the kinetics associated with the vesicle fusion process. Analysis of spike population statistics suggested detection of catecholamines from multiple release sites around the cells. The potential for such a device to be used in flow cytometry to evoke and detect exocytosis was demonstrated.
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