Improvements in the performance of an incubation-type planar patch clamp biosensor using a salt bridge electrode and a plastic (PMMA) substrate
Improvements in the performance of an incubation-type planar patch clamp biosensor using a salt bridge electrode and a plastic (PMMA) substrate
复制标题
使用盐桥电极和塑料 (PMMA) 基板改进孵化型平面膜片钳生物传感器的性能
DOI：
10.1016/j.snb.2013.12.019
复制
发表时间：
2014
影响因子：
8.4
通讯作者：
T. Urisu
中科院分区：
文献类型：
--
作者：
H. Uno;Zhi;Ysutaka Nagaoka;Noriko Takada;S. Obuliraj;Kei Kobayashi;T. Ishizuka;H. Yawo;Y. Komatsu;T. Urisu
孵化型平面膜片钳生物传感器于 2008 年首次描述，有望实现高通量神经网络筛选。在实施这项技术之前，必须克服一些技术挑战。首先，难以实现足够高的密封电阻（千兆欧姆密封），而密封电阻是膜片钳操作的重要参数。低密封阻力会产生大的基线波动，对应于固-液和液-液界面电势的波动。其次，传感器细胞必须放置在微孔中并长时间孵育，同时保持细胞活力。最后，必须开发适合高通量筛选的低成本高性能传感器芯片。这些问题在目前的工作中得到了一定程度的解决。这里开发了一种具有盐桥结构的稳定电极，以减少基线波动。通过在微流控传感器芯片中使用的塑料（聚甲基丙烯酸甲酯（PMMA））基板上使用热压印形成新颖的细胞捕获图案，改进了细胞定位技术。使用 HEK293 细胞作为模型系统，稳定电极和细胞捕获图案的组合极大地将设备成功概率从 ∼2% 提高到 60-80%。通过表征大鼠海马神经元网络来测试本结果的实用性。发现密封阻力低于从 HEK293 电池获得的密封阻力。然而，稳定电极的有效性非常明显，并且成功观察到了来自神经网络的自发通道电流。
Incubation-type planar patch clamp biosensors, first described in 2008, are expected to enable the realization of high-throughput neural network screening. Several technical challenges must be overcome before this technique may be implemented. First, it is difficult to achieve a sufficiently high seal resistance (a gigaohm seal), which is an important parameter for patch clamp operation. A low seal resistance produces large baseline fluctuations corresponding to the fluctuation of the solid–liquid and liquid–liquid interface potentials. Second, the sensor cell must be positioned at a micropore over a long incubation time while maintaining the cell viability. Finally, a low-cost high-performance sensor chip suitable for high-throughput screening must be developed. These problems have been resolved to some extent in the present work. A stable electrode with a salt bridge structure was developed here to reduce the baseline fluctuations. The cell positioning technology was improved by forming a novel cell trapping pattern using hot embossing on a plastic (polymethylmethacrylate (PMMA)) substrate for use in a microfluidic sensor chip. The combination of the stable electrode and the cell trapping pattern dramatically increased the device success probability from ∼2% to 60–80% using HEK293 cells as a model system. The utility of the present results was tested by characterizing rat hippocampal neuron networks. The seal resistance was found to be lower than the seal resistance obtained from the HEK293 cells. The effectiveness of the stable electrodes, however, was significantly clear and spontaneous channel currents from the neural network were successfully observed.
DOI：
--
发表时间：
2010
期刊：
--
影响因子：
--
作者：
B. Chow;Xue Han;A. Dobry;Xiaofeng Qian;A. Chuong;Mingjie Li;Michael A. Henninger;Gabriel M. Belfort-Gabriel-M
通讯作者：
B. Chow;Xue Han;A. Dobry;Xiaofeng Qian;A. Chuong;Mingjie Li;Michael A. Henninger;Gabriel M. Belfort-Gabriel-M
影响因子：
12.6
作者：
Pantoja, R;Nagarah, JM;Heath, JR
通讯作者：
Heath, JR