The integration of sensing devices into cell culture systems is a topic of great interest in the study of pathologies and complex biological mechanisms in real-time. In particular, the fit-for-purpose microfluidic devices called organ-on-chip (OoC), which host living engineered organs that mimic in vivo conditions, benefit greatly from the integration of sensors, enabling the monitoring of specific chemical-physical parameters that can be correlated with biological processes. In this context, copper is an essential trace element whose total concentration may be associated with specific pathologies, and it is therefore important to develop reliable analytical techniques in cell systems. Copper can be determined by using the anodic stripping voltammetry (ASV) technique, but its applicability in cell culture media presents several challenges. Therefore, in this work, the performance of ASV in cell culture media was evaluated, and an acidification protocol was tested to improve the voltammetric signal intensity. A Transwell® culture model with Caco-2 cells was used to test the applicability of the developed acidification protocol by performing an off-line measurement. Finally, a microfluidic device was designed in order to perform the acidification of the cell culture medium in an automated manner and then integrated with a silicon microelectrode to perform in situ measurements. The resulting sensor-integrated microfluidic chip could be used to monitor the concentration of copper or other ions concentration in an organ-on-chip model; these functionalities represent a great opportunity for the non-destructive strategic experiments required on biological systems under …