Action potential recording from dielectrophoretically positioned neurons inside micro-wells of a planar microelectrode array

To organise in vitro neural networks at the cellular level and study their electrical patterns, we have fabricated 4 × 4 planar microelectrode arrays using conventional photolithography. The electrode sites of these arrays are located inside micro-wells, for confining the neurons, which are connected with neighbouring wells via micro-trenches capable of guiding the outgrowth of neurites. In order to load a single neuron inside each micro-well, a simple system has been developed that utilises the phenomenon of dielectrophoresis. It operates by moving neurons towards each electrode site of an array using a dielectrophoretic force, checking for the presence of a neuron inside each micro-well using image processing, and stopping the dielectrophoretic force when detecting a neuron inside a micro-well in order to prevent more cells from being trapped. This system provides a fast, effective and inexpensive way to assemble neural grids consisting of contacts between electrodes and single neurons, as the use of micromanipulator guided micropipettes can be avoided. Spontaneous and evoked action potentials from trapped neurons were successfully recorded using a 16-channel acquisition/stimulation unit.