Fig. 2

Time course of live brain organoid imaging using microplate inserts. a Images were taken every ~ 3.5 days using an InCell Analyzer microscope using a 2.1x objective. Control refers to organoids grown without microplate inserts. Organoids grown with inserts #1 to #3 maintain their orientation and produce characteristic cortical structures (yellow arrows). Panels were made using cropped images (same size for all time points and conditions) placed on a black background. Image’s intensity levels were contracted (same histogram contraction applied to panels from all time points across all conditions) from their original 16-bit range to 8-bit RGB range for Figure preparation in Adobe Illustrator. Scale Bar = 1 mm. b Percentage of EBs that successfully matured to brain organoids under different conditions was determined by fluorescence microscopy at day 50. Successfully grown organoids were identified by an increase in size over time and identification of characteristic cortical structures. Data are means ± SEM for three independent experiments. c Organoid 2D projected area (Area) vs time plots for brain organoid cultures in the presence or absence of microplate inserts. Data points correspond to mean values for three independent experiments (n = 3), and for each experiment at least 3 organoids per condition were analysed. d Results of linear regression analysis (Area = initial organoid size + Growth rate * Days) of plots in c. The table shows the p values for the comparison of organoid growth rates between the control and microplate insert groups. Non-Significant (n.s) and Significant (*, p < 0.05)