Fifty high-content light sheet fluorescence microscopy datasets of Tribolium castaneum embryogenesis

The red flour beetle (Tribolium castaneum) is a key model organism in developmental biology, genetics, and agricultural research. To address the limited availability of high-quality microscopy data documenting its embryonic morphogenesis, we assembled the first Systematic Live Imaging Collection of Embryogenesis (SLICE-1), a large open-access resource for Tribolium embryogenesis. Using light sheet fluorescence microscopy, we acquired fifty high-content datasets totaling 200 days of recording time, 5.7 million images, and 2.7 Terabytes of data. Imaging employed five novel transgenic lines expressing mEmerald-labeled nanobodies that target distinct intracellular structures under control of the consecutively and ubiquitously active tubulin alpha 1-like protein promoter. Each line encompasses multiple sublines representing independent genomic insertions of the same transgene. Embryos were imaged in triplicates under standardized conditions, and all but three specimens developed into healthy, fertile adults, confirming the minimal invasiveness of the experimental procedure. SLICE-1 enables quantitative and comparative analyses of embryonic development, provides an overview of nanobody functionality in insects, and serves as a training resource for image processing, machine learning, and neural networks.