Tracking the lateral position of single cells and particles plays an important role in evaluating the efficiency of microfluidic cell focusing, separation and sorting. In this work, we present an N-shaped electrode-based microfluidic impedance cytometry device for the measurement of the lateral position of single cells and particles in continuous flows. Specifically, a simple analytical expression for determining the particle lateral position is derived from the measured electrical signal and geometry relationship among the positions of the flowing particles, electrodes and microchannel. This microfluidic system is experimentally validated by measuring the lateral positions of 5, 7 and 10 μm diameter beads and human red blood cells (RBCs) flowing in a 200 μm wide channel at varying flow rates up to 59.3 μl min-1. Statistical analyses show a good correlation (R2 = 0.99) and agreement (Bland-Altman analysis) between our results and those obtained by a microscopy imaging method. The resolution of our system reflected by the root-mean-square deviation (RMSD) is 10.3 μm (5.15% of the channel width) for 5 and 10 μm beads, and 11.4 μm (5.7% of the channel width) for RBCs at a flow rate of 42.4 μl min-1. Compared to the existing impedance-based methods for measuring the particle lateral position, we achieve the highest resolution, highest flow rate and smallest measured particle size (3.6 μm beads). The experimental results of the mixture with 5 and 10 μm beads demonstrate that our device does not merely measure the lateral position of single particles or cells, but also can characterize their physical properties (e.g., size) simultaneously. Furthermore, we demonstrate the position monitoring of sheath flow-induced particle focusing, which is in quantitative agreement with the results by imaging quantification. With the advantages of rapid and accurate processing of electrical signal and high throughput of the impedance flow cytometry, this novel N-shaped electrode-based system can be easily integrated with other microfluidic platforms as a downstream approach for the real-time measurement of the lateral position and physical properties of single cells and particles.