Human induced pluripotent stem cell (hiPSC)-derived neural models have emerged as invaluable tools for studying neurological disorders, such as epilepsy, Alzheimer’s, and Parkinson’s disease.Real-time, label-free measurement of electrical activity in self-organizing in vitro cellular models provides critical insight into the complexity of their neuronal networks. High-density microelectrode arrays (HD-MEAs) enable non-invasive electrophysiological recordings from various electrogenic samples, including iPSC-derived neurons, retinal explants, brain slices, and neural organoids.In this study, we used MaxOne and MaxTwo high-density MEA platforms (MaxWell Biosystems AG, Switzerland), with 26,400 electrodes per well, to record extracellular action potentials in neural organoids at different scales, ranging from cell population networks to single-cell resolution and subcellular levels. We showcased the flexible selection of electrodes for recording neural activity, increasing the reproducibility and statistical power of the data collected. Key metrics such as firing rate, spike amplitude, and network burst profile were extrapolated in a parallelized manner to capture even the smallest neuronal signals.Furthermore, we characterized axonal function and structure using the AxonTracking Assay, which allows measurement of action potential conduction velocity, latency, axonal length, and branching. This automated assay facilitates high-throughput characterization of disease models targeting axon initial segments, axonal branching, development, and conduction.MaxWell Biosystems’ HD-MEA platforms, along with automatically generated plots and extracted metrics, provide a unique, user-friendly approach to identifying and isolating functionally active regions in 3D cultures. These powerful platforms enable long-term in vitro disease modeling and compound testing in acute recordings and/or longitudinal studies.