Stepwise pattern modification of neuronal network in photo-thermally-etched agarose architecture on multi-electrode array chip for individual-cell-based electrophysiological measurement
Stepwise pattern modification of neuronal network in photo-thermally-etched agarose architecture on multi-electrode array chip for individual-cell-based electrophysiological measurement
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DOI：
10.1039/b406885h
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发表时间：
2005-01-01
期刊：
影响因子：
6.1
通讯作者：
Yasuda, K
中科院分区：
文献类型：
--
作者：
Suzuki, I;Sugio, Y;Yasuda, K
我们已经开发了一个程序，使用单个细胞为基础的芯片上的多电极阵列（MEA）细胞培养系统与琼脂糖微室（AMC）阵列相邻的活神经元之间的网络模式和神经突连接方向的逐步拓扑控制。该过程能够灵活和精确地控制细胞位置，并通过逐步光热蚀刻容易和灵活地控制AMC中细胞之间连接的图案修饰，其中即使在培养期间，芯片上的琼脂糖层的一部分也用1480 nm红外激光束熔化。在适当的激光功率和这种分步过程中，我们可以在培养过程中以分步方式制造狭窄的微米级凹槽（微通道）。使用这个程序，我们选择性地控制轴突和树突的伸长方向，并通过免疫染色确认方向。我们还证明了电生理信号的单向传输对齐海马神经元中的神经突连接的方向控制使用这种逐步光热蚀刻程序。这些结果证明了在片上AMC/MEA细胞培养系统中使用逐步光热蚀刻逐个形成微通道来全方向控制神经元之间的神经突连接的潜力。因此，我们可以更好地理解神经元网络模式和连接方向的意义。
We have developed a procedure for stepwise topographical control of network patterns and neurite connection directions between adjacent living neurons using an individual-cell-based on-chip multi-electrode array ( MEA) cell cultivation system with an agarose microchamber (AMC) array. This procedure enables flexible and precise control of the cell positions and easy and flexible control of the pattern modification of connections between the cells in AMCs through stepwise photo-thermal etching in which a portion of the agarose layer on the chip is melted with a 1480 nm infrared laser beam even during cultivation. With adequate laser power and this stepwise procedue, we can fabricate narrow micrometer-order grooves (microchannels) during cultivation in a stepwise manner. Using this procedure, we controlled the direction of elongation of axons and dendrites selectively and confirmed the direction by immunostaining. We also demonstrated electrophysiological one-way transmission of signals among aligned hippocampal neurons in which the directions of the neurite connections were controlled using this stepwise photo-thermal etching procedure. These results demonstrate the potential of full direction control of neurite connections between neurons using stepwise photo-thermal etching to form microchannels one by one in an on-chip AMC/MEA cell cultivation system. We can thus better understand the meaning of neuronal network patterns and connection directions.