Extended Data Fig. 1: Protocol development for organoids compatible with long-term live light sheet imaging.
From: Morphodynamics of human early brain organoid development
a) Schematic of the protocol developed in this study (Protocol I) and a previous protocol to develop multi-region brain organoids (Protocol II)2,26. b) Comparison of organoid size at day 4 post aggregation at two cell seeding concentrations. Scale Bar is 500 micrometers. c) Brightfield images showing organoid growth from day 8–11 in the two different protocols. N = 3, n > 3. Scale Bar is 500 micrometers. d) Heatmap showing normalized marker gene expression grouped per cluster of organoid time-course scRNA-seq data shown in Fig. 1b. e,f) Images of example organoids stained with fluorescence in situ hybridization chain reaction (HCR) showing regionalized expression of marker genes in false color, merged using Fiji. Three organoids were used for staining in each case and the n indicates the number of organoids imaged. Organoids were stained whole-mount and images were acquired as z-stacks. Selected cross-sections after reorientation of the acquired volume are shown for each organoid. e) Day 11 (top n = 1), (bottom n = 3). f) Day 15 top left, n = 2, top right n = 3, middle left n = 1, middle right n = 3, bottom left n = 3. Scale, 250 micrometers in all images.
