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A highâthroughput dielectrophoresisâbased cell electrofusion microfluidic device
- Hu, Ning, Yang, Jun, Yin, ZhengâQin, Ai, Ye, Qian, Shizhi, Svir, Irina B., Xia, Bin, Yan, JiaâWen, Hou, WenâSheng, Zheng, XiaoâLin
- Electrophoresis 2011 v.32 no.18 pp. 2488-2495
- aluminum, biocompatibility, cell fusion, electric field, electrodes, electrofusion, electrophoresis, electroporation, mammals, polyethylene glycol, protoplasts, silica, silicon
- A highâthroughput cell electrofusion microfluidic chip has been designed, fabricated on a siliconâonâinsulator wafer and tested for in vitro cell fusion under a low applied voltage. The developed chip consists of six individual straight microchannels with a 40âÎ¼m thickness conductive highly doped Si layer as the microchannel wall. In each microchannel, there are 75 pairs of counter protruding microelectrodes, between which the cell electrofusion is performed. The entire highly doped Si layer is covered by a 2âÎ¼m thickness aluminum film to maintain a consistent electric field between different protruding microelectrode pairs. A 150ânm thickness SiO2 film is subsequently deposited on the top face of each protruding microelectrode for better biocompatibility. Owing to the short distance between two counter protruding microelectrodes, a high electric field can be generated for cell electrofusion with a low voltage imposed across the electrodes. Both mammalian cells and plant protoplasts were used to test the cell electrofusion. About 42â68% cells were aligned to form cellâcell pairs by the dielectrophoretic force. After cell alignment, cell pairs were fused to form hybrid cells under the control of cell electroporation and electrofusion signals. The averaged fusion efficiency in the paired cells is above 40% (the highest was about 60%), which is much higher than the traditional polyethylene glycol method (<5%) and traditional electrofusion methods (â¼12%). An individual cell electrofusion process could be completed within 10âmin, indicating a capability of high throughput.