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We present cell electroporation chip using a single channel with multiple electric field, E, zones. The present chip, where multiple E zones are generated by a pair of external electrodes across a stepwise single channel, provides the optimal E conditions for stable cell electroporation with high viability in a single experiment. The optimal E for both H23 and A549 cells was 0.4 kV/cm with the maximum percentage of the viable and electroporated cells of 51.4 ± 3.0% and 26.6 ± 0.7%, respectively. The present cell electroporation chip has potential for use in integrated cell chips to find the optimal E conditions for the electroporation study.