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A microfluidic device integrated with 3D thin film microelectrode arrays wrapped around serpentine-shaped microchannel walls has been designed, fabricated and tested for cell electrofusion. Each microelectrode array has 1015 discrete microelectrodes patterned on each side wall, and the adjacent microelectrodes are separated by coplanar dielectric channel wall. The device was tested to electrofuse K562 cells under a relatively low voltage. Under an AC electric field applied between the pair of the microelectrode arrays, cells are paired at the edge of each discrete microelectrode due to the induced positive dielectrophoresis. Subsequently, electric pulse signals are sequentially applied between the microelectrode arrays to induce electroporation and electrofusion. Compared to the design with thin film microelectrode arrays deposited at the bottom of the side walls, the 3D thin film microelectrode array could induce electroporation and electrofusion under a lower voltage. The staggered electrode arrays on opposing side walls induce inhomogeneous electric field distribution, which could avoid multi-cell fusion. The alignment and pairing efficiencies of K562 cells in this device were 99% and 70.7%, respectively. The electric pulse of low voltage (∼9 V) could induce electrofusion of these cells, and the fusion efficiency was about 43.1% of total cells loaded into the device, which is much higher than that of the convectional and most existing microfluidics-based electrofusion devices.

A cell electrofusion microfluidic device integrated with 3D thin-film microelectrode arrays