Fig. 4From: Long-term live imaging and multiscale analysis identify heterogeneity and core principles of epithelial organoid morphogenesisVolume analysis of three representative mPOs reveals different oscillation frequencies. MPOs (seeded and maintained in one Z1-FEP-cuvettes) expressed the nuclei marker Rosa26-nTnG (grey). Organoids were imaged for 6 days and three representative organoids were analysed with our previously published nuclei segmentation pipeline [22] in regard to size oscillation events. A size oscillation lasts between 30 min and 2 h. a Volume over time for each organoid approximated from the cell nuclei segmentation. b Close-up of three (red), two (green) and seven (blue) size oscillation events with more than 5% volume reductions are shown. c Typical images of organoid size oscillation. The upper row shows the nuclei in grey in the raw image, the lower row the segmented cell nuclei. Each colour illustrates a single-cell nucleus. Microscope: Zeiss Lightsheet Z.1; objective lenses: detection: W Plan-Apochromat × 20/1.0, illumination: Zeiss LSFM × 10/0.2; laser lines: 561 nm; filters: laser block filter (LBF) 405/488/561; voxel size: 1.02 × 1.02 × 2.00 μm3; recording interval: 30 min; scale bar: 100 μmBack to article page