Cell surface charge is an important element of the function of biological barriers, but no chip device has been described to measure cell surface charge properties of confluent barrier cell monolayers. The aim of this study was the design and fabrication of a dynamic lab-on-a-chip (LOC) device which is suitable to monitor transcellular electrical resistance, as well as streaming potential parallel to the surface of cell layers. We successfully measured the streaming potential of a biological barrier culture model with the help of our previously published versatile lab-on-a-chip device equipped with two Ag/AgCl electrodes. The inclusion of these "zeta electrodes", a voltage preamplifier and an oscilloscope in our set-up made it possible to successfully record signals describing the surface charge properties of brain endothelial cell monolayers, used as a barrier model in our experiments. Data obtained on the new chip device were verified by comparing streaming potential results measured in the LOC device and zeta potential results by the commonly used laser-Doppler velocimetry (LDv) method and model simulations. Changes in the negative surface charge of the barrier model by treatments with neuraminidase enzyme modifying the cell membrane glycocalyx or lidocaine altering the lipid membrane charge could be measured by both the upgraded LOC device and LDv. The new chip device can help to gain meaningful new information on how surface charge is linked to barrier function in both physiological and pathological conditions.