The prostate biopsy method shows a high false negative result because the suspicious tissue considered as cancer is not confirmed during tissue sampling. Thus, repeated biopsy procedures and diagnostic errors in relation to prostate cancer frequently occur. The purpose of this research is to enhance the prostate cancer detection rate by using microfluidic electrical impedance spectroscopy (lEIS), which allows real-time measurement of the electrical impedance of a single human prostate normal cell and cancer cell. The lEIS was equipped with a movable flexible membrane, which is operated by pneumatic pressure to capture the single cell on the surface of sensing electrodes. The forced tight contact between the cell and electrodes makes it possible to measure the electrical characteristics of the cell with a high sensitivity. The lEIS discriminates well between normal human prostate cells (RWPE-1) and cancer cells (PC-3) at 8.7 kHz based on the electrical signal responses of the cells. The average difference rates of admittance magnitude and susceptance are 54.55% and 54.59%, respectively. The developed lEIS also shows high repeatability, which was verified by a deionized water test conducted before and after each cell assay; the maximum variance of both the impedance and admittance at 8.7 kHz was as small as 9.48%. V C 2013 AIP Publishing LLC. [http://dx