In this study, a novel method for detecting bacterial cells in deionized (DI) water suspension is presented by using fluidic electrodes with a hydrodynamic focusing technique. KCl solution was utilized as both sheath flow and fluidic electrodes, and the bacterial suspension was squeezed to form three flowing layers with different conductivities on a microfluidic chip. An impedance analyzer was connected with the KCl solution through two Ag/AgCl wires to apply an AC voltage to fluidic layers within a certain frequency for impedance measurements. Porphyromonas gingivalis and Escherichia coli were detected and linear relationships were found between the impedance and the logarithmic value of the bacterial concentration in certain cell concentration ranges. It is demonstrated that bacterial detection using the microdevice is rapid and convenient, with a chip made of simple flow channels, and the detection sensitivity of cell counting can be tuned by varying the width of the sample flow layer through changing input velocities, showing a detection limit of 10(3) cells mL(-1).