Dissociated neurons cultured in vitro can serve as a model system for studying the dynamics of neural networks. Such studies depend on techniques for stimulating patterns of neural activity. We show a technique for extracellular stimulation of dissociated neurons cultured on silicon wafers. When the silicon surface is reverse biased, electrical current can be generated near any neuron by pulsing a laser. Complex spatiotemporal stimulation patterns can be produced by directing a single beam with an acousto-optic deflector. The technique can generate a stimulating current at any location in the culture. This contrasts with multielectrode arrays (MEAs), which can stimulate only at fixed electrode locations. To characterize reliability and spatial selectivity of stimulation, we used intracellular (patch-clamp) recordings to monitor the effect of targeted laser pulses on cultured hippocampal neurons. Action potentials could be stimulated with submillisecond precision and 100-micron spatial resolution at rates exceeding 100 Hz. Optimal control parameters for stimulation are discussed.
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