Millifluidic triMEA organoid platform with stretchable microelectrode arrays for parallel organoid electrophysiology under standardized perfusion

Organoids enable rigorous studies of neurodevelopment and disease, yet existing platforms often compromise between precise microenvironment control and reliable electrophysiological measurement. The challenge of achieving uniform perfusion alongside multi-well volumetric neural recordings limits parallel functional characterization and technical reproducibility across samples. Here, we present the triMEA millifluidic platform, which integrates three stretchable microelectrode arrays within a single device and delivers uniform perfusion across three independent wells. A single inlet supplies identical perfusion to each well, which stabilizes flow patterns and equalizes mechanical stimuli between chambers. This design facilitates simultaneous, independent electrophysiological recordings from three organoids within a single experimental setup. Using human midbrain organoids as a proof-of-concept biological model, we provide computational and experimental validation of the fluidic profiles and demonstrate electrophysiological recordings across all three wells under continuous perfusion. This platform establishes a reproducible and standardized foundation for parallel organoid interfacing.