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

Abstract Over the past decade, air-liquid interface cerebral organoids (ALI-COs) have emerged as powerful in vitro models that capture essential structural and functional traits of the human brain, offering an exciting alternative to traditional animal models in neuroscience. Yet, the full potential of these systems has remained untapped due to the lack of non-invasive, long-term electrophysiological tools capable of preserving organoid integrity. Existing techniques, ranging from patch clamping to rigid and 3D microelectrode arrays, often compromise organoid growth and disrupt delicate cytoarchitecture. Here, we present NeuroSuite, an innovative bioelectronic platform designed to overcome these challenges. At its core is Neuroweb, a perforated, ultra-thin, and conformable organic microelectrode array engineered for minimal disruption of nutrient and oxygen exchange. Neuroweb is reusable and supports stable recordings for over six months, making it uniquely suited for longitudinal studies. Coated with poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS), a high-performance mixed ionic-electronic conductor, Neuroweb delivers exceptional signal-to-noise ratio recordings with high spatial precision. By pairing Neuroweb with NeuroMaps, an intuitive software for interactive analysis and visualisation, NeuroSuite enables long-term, non-invasive tracking and spatial mapping of electrical activity from brain organoids and ex vivo brain slices at the air-liquid interface. Following rigorous validation, we demonstrate that NeuroSuite can capture both high- and low-frequency throughout maturation. Our pipeline reveals evolving network connectivity, including the development of GABA-ergic interneurons, and concurrent shifts in high-frequency spiking and low-frequency oscillations indicative of a refinement in the excitatory-inhibitory balance. Finally, automated data acquisition and spatial spike mapping highlight local activity changes in response to media composition, a factor often overlooked in conventional recordings. NeuroSuite thus opens a new frontier in organoid neuroscience, enabling precise, long-term monitoring essential for modelling neurological diseases, understanding human brain development, and accelerating drug discovery. Teaser Conformal organic bioelectronic arrays, combined with an open-access toolbox for analysis and visualisation, reveal real-time and long-term neural dynamics in brain organoid slices at the air-liquid interface. Competing Interest Statement NeuroSuite, including both the device and monitoring toolbox, has been patented (United Kingdom Patent Application No. 2511343.2).