We propose a microfluidic platform with integrated microfabricated electrodes for real-time impedance profiling of transwell-based barrier models, which can be subjected to dynamic microfluidic flow. This platform overcomes the limitations of conventional methods that are based on invasive permeability assays and single time-point impedance measurements: It enables continuous, non-invasive monitoring of tissue barrier integrity at high spatial and temporal resolution. We demonstrate the capabilities of our system by continuously monitoring the gradual loss of barrier integrity in upper-airway-tissue models exposed to non-physiological liquid-liquid interface conditions.