NeuroSuite for Long-Term Functional and Structural Studies of Air-Liquid Interface Cerebral Organoids

Long-term electrophysiological monitoring of human cerebral organoids remains challenging, as 3D tissues rapidly develop necrotic cores once diffusion limits are exceeded. Air-liquid interface cerebral organoids (ALI-COs) overcome this by maintaining thin slices with continuous metabolic exchange, preserving viability, yet no system allows stable, in situ recordings over months. Here, we introduce NeuroSuite, a modular platform for long-term, spatially resolved neural monitoring of ALI-COs. Its core, Neuroweb, is an ultrathin, perforated microelectrode array that preserves cellular diversity, evidenced by immunofluorescence and RNA sequencing, while enabling low-impedance (∼25 kΩ at 1 kHz) recordings. Complementing this, NeuroMaps, our open-source analysis suite, streamlines signal quality control, spike and field potential analysis, and integration with imaging. NeuroSuite is suitable for capturing maturation-dependent network dynamics, including potential excitation-inhibition shifts and oscillatory activity, beyond 180 days. Acute ex vivo rat brain slice recordings further demonstrate its versatility, establishing NeuroSuite as a robust platform for developmental neuroscience, disease modelling, and drug screening.