Individual-Cell-Based Electrophysiological Measurement of a Topographically Controlled Neuronal Network Pattern Using Agarose Architecture with a Multi-Electrode Array

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Abstract
We have developed a new type of individual-cell-based electrophysiological measurement method using an on-chip multi-electrode array (MEA) cell-cultivation system with an agarose microchamber (AMC) array for topographical control of the network patterns of a living neuronal network. The advantages of this method are that it allows the recording of the firing of multiple cells simultaneously for weeks without contamination using the MEA, and that it allows control of the cell positions and numbers, and their connections for cultivation using AMCs with microchannels fabricated by photothermal etching where a portion of the agarose layer is melted with a 1480 nm infrared laser beam. Using this method, we formed an individual-cell-based neural network pattern of Rat hippocampal cells within the AMC array without cells escaping from the electrode positions in the microchamber during a thirteen-day cultivation, and could record the cell firing of lined-up hippocampal cells in response to 20 µA, 5 kHz stimulation via an electrode. This demonstrated the potential of our on-chip AMC/MEA cell cultivation method for long-term single-cell-based electrophysiological measurement of a neural network system for understanding the topographical meaning of neuronal network patterns.

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