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We performed single pulses current measurement on single cells using dual nanoprobe through environmental scanning electron microscope nanomanipulator system. The ability to characterize the electrical property of single cells can be used as a novel method for cell viability detection in quantitative and instantaneous manners. The nanoprobe was successfully fabricated using focused ion beam tungsten deposition and etching processes. The characteristics of the nanoprobe were examined from the energy dispersion spectrometry and noise analyses. In this paper, for the first time, the electrical property of single cells under their native condition was presented. In order to apply this method for cell viability detection, two types of cells were used, i.e., dead cells and live cells. The results showed that there is a significant difference on the electrical measurement data between dead and live cells.