Sixteen isotropic 3D fluorescence live-imaging datasets of gastrulation in the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae).

Gastrulation is a pivotal phase of early embryogenesis during which initially uniform cells undergo coordinated movements and fate specification to establish the basic body plan. Although previous studies have advanced our understanding of gastrulation in the red flour beetle Tribolium castaneum, quantitative morphogenetic data remain limited. Here, we present the second Systematic Live Imaging Collection of Embryogenesis (SLICE-2), consisting of sixteen isotropic 3D fluorescence live imaging datasets that document gastrulation and early germband elongation in Tribolium. Imaging was performed using light sheet fluorescence microscopy in conjunction with a homozygous transgenic line that expresses mEmerald-labeled nanobodies against histone H2A/H2B. Embryos were recorded along four directions, and the resulting axial image stacks were subjected to multiview fusion to obtain isotropic 3D images of entire embryos with reduced shadowing artifacts. The collection includes weighted-average fusion and fusion-deconvolution derivatives, and nuclei segmentation data are available for selected developmental stages. SLICE-2 expands the microscopy data resources for Tribolium, enabling comprehensive analyses of gastrulation dynamics while also providing benchmark data for image processing, segmentation, and modeling approaches.