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

ABSTRACT 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.