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A water compartment cell culture lid enables stable longitudinal recording of neuronal networks in vitro

Maurer, B.; Fischer, F.; Amos, G.; Vasiliauskaite, V.; Vörös, J.

2026-04-05 neuroscience
10.64898/2026.04.01.713917 bioRxiv
Show abstract

Longitudinal electrophysiological recordings of neuronal networks are essential for studying network maturation, plasticity, and pharmacological responses. Yet current microelectrode array (MEA) approaches are limited by evaporation-induced drift in culture conditions, exacerbated by heat dissipation from active recording electronics on CMOS-based high-density MEAs. We present a cell culture lid featuring a water compartment at its interface that eliminates evaporation whilst maintaining gas exchange. Combined with a custom incubator that uses independent temperature control of the MEA to prevent condensation, the system enables stable, un-interrupted recordings for weeks. We show that perturbations in firing rate and functional connectivity following medium exchange are significantly reduced by suppressing evaporation. We demonstrate continuous 35-day recordings of patterned human iPSC-derived neuronal networks with a single medium exchange, revealing the spontaneous emergence and consolidation of spatiotemporal firing patterns during maturation. All design files are provided to facilitate adoption across culturing platforms, enabling un-interrupted longitudinal interfacing with network dynamics for studies of plasticity, chronic pharmacology, and developmental trajectories in individual cultures.

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