Assembly of imaging chambers and high-resolution imaging of early chick embryos

The chick embryo is a classical model system for developmental biology. Its flat disk morphology exemplifies the amniote embryos from reptiles, birds, and most mammals (including humans). It is also particularly amenable to manipulation and culture both in vitro and in ovo. As a consequence, studies with this system have been important for promoting our understanding of cell and tissue movements, lineages, inductive processes, and cell fate decisions. In recent years, imaging techniques have been devised that allow these questions to be studied at the cellular level at late neurulation and segmentation stages. Modern improvements to imaging techniques now allow direct visualization of molecular interactions and cellular events and are likely to be instrumental in linking these processes to whole tissue and embryo morphogenesis. Here, we describe a method for high-resolution (confocal or multiphoton) time-lapse imaging of chick embryos at stages spanning pregastrulation to early neurulation. We give detailed instructions for assembling the specialized imaging chambers required for this procedure.