Design rule for optimization of microelectrodes used in electric cell-substrate impedance sensing (ECIS)

This paper presents an experimentally derived design rule for optimization of microelectrodes used in electric cell-substrate impedance sensing (ECIS) up to 10 MHz. The effect of change in electrode design (through electrode sensor area, lead trace widths, and passivation coating thickness) on electrode characteristics was experimentally evaluated using electrochemical impedance spectroscopy (EIS) measurements and analyzed using equivalent circuit models. A parasitic passivation coating impedance was successfully minimized by designing electrodes with either a thicker passivation layer or a smaller lead trace area. It was observed that the passivated lead trace area to passivation coating thickness ratio has a critical value of 5.5, under which the impedance contribution of the coating is minimized. The optimized design of ECIS-based microelectrode devices reported in this work will make it possible to probe the entire beta dispersion region of adherent biological cell layers by reducing measurement artifacts and improving the quality of data across the beta-dispersion region. The new design will enable the use of the commonly used ECIS technique to measure real-time cellular properties in high frequency ranges (beta dispersion) that was not possible thus far.