In total, eight independent neural differentiations were tested per patient, 210 movies of spontaneous calcium transients (110 control and 100 SCZD) were generated and 2,676 ROIs (1,158 control and 1,518 SCZD ROIs) were analyzed. Up to four 90-second videos of Fluo-4AM fluorescence were recorded per neural differentiation per patient with a spinning disc confocal microscope at 28 frames per second (SI Fig. 5A). Using ImageJ software, regions of interest (ROIs) can be manually selected and the mean pixel intensity of each ROI can be followed over time, generating time trace data for each ROI. The data were analyzed in Matlab where background subtraction was performed by normalizing traces among traces of the sample, and spike events were identified based on the slope and amplitude of the time trace.
