A novel high-aspect-ratio penetrating microelectrode array was designed and fabricated for the purpose of recording neural activity. The array allows two dimensional recording of 64 sites in vitro with high aspect ratio penetrating electrodes. Traditional surface electrode arrays, although easy to fabricate, do not penetrate to the viable tissue such as central hippocampus slices and thus have a lower signal/noise ratio and lower selectivity than a penetrating array. In the unfolded hippocampus preparation, the CA1-CA3 pyramidal cell layer in the whole unfolded rodent hippocampus preparation is encased by the alveus on one side and the Schaffer tract on the other and requires penetrating electrodes for high signal to noise ratio recording. An array of 64 electrode spikes, each with a target height of 200μm and diameter of 20μm, was fabricated in silicon on a transparent glass substrate. The impedance of the individual electrodes was measured to be approximately 1.5MΩ±497kΩ. The signal to noise ratio was measured and found to be 19.4±3dB compared to 3.9±0.8dB S/N for signals obtained with voltage sensitive dye RH414. A mouse unfolded hippocampus preparation was bathed in solution containing 50 micro-molar 4-amino pyridine and a complex two dimensional wave of activity was recorded using the array. These results indicate that this novel penetrating electrode array is able to obtain data superior to that of voltage sensitive dye techniques for broad field two-dimensional neuronal activity recording. When used with the unfolded hippocampus preparation, the combination forms a uniquely capable tool for imaging hippocampal network activity in the entire hippocampus.
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