Trans-epithelial electrical resistance (TEER) is a widely used as an experimental readout and a quality control assay for measuring the integrity of epithelial monolayers. However, it is difficult to compare TEER values among different Organ-on-a-Chip (OoC) devices due to considerable errors in measurements. Here we report an OoC design method focusing on the electrodes that achieves the accurate measurement of TEER values. This approach has employed an index of “sensitivity variation” calculated from current density depending on electrodes configuration and structure of cell culture chamber, allowing to identify the suitable electrodes design for TEER measurements. The validity of this approach was successfully confirmed to measure TEER by impedance spectroscopy measurements and detect changes in integrity of Caco-2 cell. The present approach will allow a comparison of TEER values across any OoC as well as real-time assessment of biological functions.