Single cell and neural process experimentation using laterally applied electrical fields between pairs of closely apposed microelectrodes with vertical sidewalls.
Single cell and neural process experimentation using laterally applied electrical fields between pairs of closely apposed microelectrodes with vertical sidewalls.
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DOI：
10.1016/j.bios.2009.05.024
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发表时间：
2009-08-15
影响因子：
12.6
通讯作者：
Sretavan DW
中科院分区：
文献类型：
--
作者：
Chang WC;Sretavan DW
随着生物医学研究越来越多地转向单细胞和亚细胞结构的实验，需要能够在相应的小范围内进行操作和刺激的微型设备。我们提出了一种微电极阵列（MEA），其特点是加厚的微电极具有垂直侧壁（VSW），可以将电场水平聚焦在位于成对电极之间的目标上。这些微电极是用SU-8光刻胶模压成型的金电镀而成的。有限元模拟表明，与传统平面微电极相比，VSW电极产生的电场更加均匀。使用3µm厚、间隔10µm的配对微电极，我们能够对单个轴突过程进行局部电穿孔，正如EGTA进入轴突内局部螯合钙，猝灭预加载钙指示剂染料的荧光所证明的那样。使用相同的电极配置对单个细胞进行电穿孔，从而靶向转染表达细胞质可溶性绿色荧光蛋白（GFP）的转基因。除了电促动外，我们的电极配置还能够对单个神经元进行精确的定向场刺激，从而产生可以通过光学手段跟踪的动作电位。凭借其提供良好表征电场的能力和通用性，我们的成对VSW电极配置可能为神经科学和生物医学研究的广泛领域的高通量和高含量实验提供新工具的基础。
As biomedical research has moved increasingly towards experimentation on single cells and subcellular structures, there has been a need for microscale devices that can perform manipulation and stimulation at a correspondingly small scale. We propose a microelectrode array (MEA) featuring thickened microelectrodes with vertical sidewalls (VSW) to focus electrical fields horizontally on targets positioned in between paired electrodes. These microelectrodes were fabricated using gold electroplating that was molded by photolithographically patterned SU-8 photoresist. Finite element modeling showed that paired VSW electrodes produce more uniform electrical fields compared to conventional planar microelectrodes. Using paired microelectrodes, 3µm thick and spaced 10µm apart, we were able to perform local electroporation of individual axonal processes, as demonstrated by entry of EGTA to locally chelate intra-axonal calcium, quenching the fluorescence of a pre-loaded calcium indicator dye. The same electrode configuration was used to electroporate individual cells, resulting in the targeted transfection of a transgene expressing a cytoplasmically soluble green fluorescent protein (GFP). In addition to electropration, our electrode configuration was also capable of precisely targeted field stimulation on individual neurons, resulting in action potentials that could be tracked by optical means. With its ability to deliver well-characterized electrical fields and its versatility, our configuration of paired VSW electrodes may provide the basis for a new tool for high-throughput and high-content experimentation in broad areas of neuroscience and biomedical research.
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影响因子：
3.9
作者：
Barbier, Valessa;Tatoulian, Michael;Tabeling, Patrick
通讯作者：
Tabeling, Patrick
影响因子：
14.8
作者：
Kaech, Stefanie;Banker, Gary
通讯作者：
Banker, Gary
影响因子：
2.4
作者：
LOEW, LM;COHEN, LB;WU, JY
通讯作者：
WU, JY
影响因子：
6.1
作者：
Ravula, Surendra K.;McClain, Maxine A.;Frazier, A. Bruno
通讯作者：
Frazier, A. Bruno
DOI：
10.1021/la8021479
发表时间：
2008-11-18
期刊：
Langmuir : the ACS journal of surfaces and colloids
影响因子：
--
作者：
Chang WC;Sretavan DW
通讯作者：
Sretavan DW