Long-term fluorescence live imaging of Tribolium castaneum embryos: principles, resources, scientific challenges and the comparative approach

Light sheet-based fluorescence microscopy became an important tool in insect developmental biology due to its high acquisition speed, low photo-bleaching rate and the high survival probability of the specimens. Initially applied to document the embryogenesis of Drosophila melanogaster, it is now used to investigate the embryonic morphogenesis of emerging model organisms such as the red flour beetle Tribolium castaneum. Here, we discuss the principles of light sheet-based fluorescence microscopy and outline Tribolium as a model organism for developmental biology. We summarize labeling options and present two custom-made transgenic lines suitable for live imaging. Finally, we highlight studies on Tribolium that address scientific questions with fluorescence live imaging and discuss the comparative approach to investigate insect morphogenesis in an evolutionary context.