Microwell device for targeting single cells to electrochemical microelectrodes for high-throughput amperometric detection of quantal exocytosis.
Microwell device for targeting single cells to electrochemical microelectrodes for high-throughput amperometric detection of quantal exocytosis.
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DOI：
10.1021/ac1033616
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发表时间：
2011-04-01
影响因子：
7.4
通讯作者：
Gillis, Kevin D.
中科院分区：
文献类型：
--
作者：
Liu, Xin;Barizuddin, Syed;Shin, Wonchul;Mathai, Cherian J.;Gangopadhyay, Shubhra;Gillis, Kevin D.
电化学微电极通常用于检测安培电流的尖峰，所述尖峰对应于来自单个囊泡的可氧化递质的胞吐，即，量子胞吐作用我们正在研制一种在微芯片上的透明多电化学电极阵列，以实现量子胞吐的自动化测量。在这里，我们报告了一种改进的设备，以针对单个细胞的阵列中的每个微电极的发展。使用SU-8光致抗蚀剂中制造的细胞大小的微孔陷阱以及与电极配准的聚（L-赖氨酸）图案化以促进细胞粘附来实现有效的靶向（~75%）。使用聚（乙二醇）使电极之间的表面抗细胞粘附，以促进细胞移动到电极“对接位点”。我们使用测试分析物铁氰化物证明电极的活性，并在设备上记录牛肾上腺嗜铬细胞的量子胞吐。单个器械上的多个细胞记录证明了尖峰测量的一致性，来自相同电极的多个记录证明了该器械可以清洁和重复使用，而不会降低性能。新设备将使量子胞吐作用的高通量研究成为可能，也可能应用于快速筛选药物或毒素对胞吐作用的影响。
Electrochemical microelectrodes are commonly used to detect spikes of amperometric current that correspond to exocytosis of oxidizable transmitter from individual vesicles, i.e., quantal exocytosis. We are developing transparent multi- electrochemical electrode arrays on microchips in order to automate measurement of quantal exocytosis. Here we report development of an improved device to target individual cells to each microelectrode in an array. Efficient targeting (~75%) is achieved using cell-sized micro-well traps fabricated in SU-8 photoresist together with patterning of poly (L-lysine) in register with electrodes to promote cell adhesion. The surface between electrodes is made resistant to cell adhesion using poly (ethylene glycol) in order to facilitate movement of cells to electrode “docking sites”. We demonstrate the activity of the electrodes using the test analyte ferricyanide and perform recordings of quantal exocytosis from bovine adrenal chromaffin cells on the device. Multiple cell recordings on a single device demonstrate the consistency of spike measurements and multiple recordings from the same electrodes demonstrate that the device can be cleaned and reused without degradation of performance. The new device will enable high-throughput studies of quantal exocytosis and may also find application in rapidly screening drugs or toxins for effects on exocytosis.
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影响因子：
6.1
作者：
Dittami GM;Rabbitt RD
通讯作者：
Rabbitt RD
影响因子：
3
作者：
Colliver, TL;Hess, EJ;Ewing, AG
通讯作者：
Ewing, AG
DOI：
10.1098/rstb.1999.0382
发表时间：
1999-02-28
影响因子：
6.3
作者：
Oheim, M;Loerke, D;Stühmer, W
通讯作者：
Stühmer, W
影响因子：
64.8
作者：
CHOW, RH;VONRUDEN, L;NEHER, E
通讯作者：
NEHER, E
影响因子：
56.9
作者：
Lee, Haeshin;Dellatore, Shara M.;Messersmith, Phillip B.
通讯作者：
Messersmith, Phillip B.