Fig 1: Robotic Scalable Neuronal Differentiation(A) Overview of neuronal differentiation strategy compatible with automation.(B) Phase-contrast image showing a typical neuronal culture (day 30; magnification 20x).(C) Neuronal cells develop highly dense network of neurites upon further maturation (day 50; magnification 40x).(D) Immunocytochemical analysis showing cortical neurons expressing TUJ1 and CUX1 (magnification, 20x).(E) Immunocytochemical analysis demonstrating the presence of cortical neurons expressing MAP2 and CTIP2 (magnification, 20x).(F) Majority of cells express vGLUT1, a marker for excitatory neurons (magnification, 20x).(G) Example of neuronal cells showing immunoreactivity for the inhibitory neurotransmitter GABA (magnification, 63x)(H) Robotic MEA platform used for high-throughput electrophysiology and functional cell characterization.(I) Spontaneous activity of hiPSC-derived neurons after 6 weeks of differentiation as measured by MEA.(J) Overlay plot of 10 spikes detected from one channel of a representative MEA recording to demonstrate similarity between spikes detected.