Abstract
Studies of the real-time dynamics of embryonic development require a gentle embryo handling method, the possibility of long-term live imaging during the complete embryogenesis, as well as of parallelization providing a population’s statistics, while keeping single embryo resolution. We describe an automated approach that fully accomplishes these requirements for embryos of Caenorhabditis elegans, one of the most employed model organisms in biomedical research. We developed a microfluidic platform which makes use of pure passive hydrodynamics to run on-chip worm cultures, from which we obtain synchronized embryo populations, and to immobilize these embryos in incubator microarrays for long-term high-resolution optical imaging. We successfully employ our platform to investigate morphogenesis and mitochondrial biogenesis during the full embryonic development and elucidate the role of the mitochondrial unfolded protein response (UPRmt) within C. elegans embryogenesis. Our method can be generally used for protein expression and developmental studies at the embryonic level, but can also provide clues to understand the aging process and age-related diseases in particular.
Highlights
Studies of the real-time dynamics of embryonic development require a gentle embryo handling method, the possibility of long-term live imaging during the complete embryogenesis, as well as of parallelization providing a population’s statistics, while keeping single embryo resolution
Whether conditions in the embryonic phase of life have an influence on the later development is a much more challenging question to answer, mainly because systematic C. elegans embryonic morphogenesis studies are still difficult from a technical point of view
To enable systematic analysis of C. elegans embryonic morphogenesis, we developed a microfluidic platform for automated on-chip worm culture, creation of synchronized embryo arrays, and for long-term parallel live imaging at the single embryo level
Summary
Studies of the real-time dynamics of embryonic development require a gentle embryo handling method, the possibility of long-term live imaging during the complete embryogenesis, as well as of parallelization providing a population’s statistics, while keeping single embryo resolution. “Worm-chips” have successfully demonstrated their high potential at enhancing worms’ handling and accurate imaging, for applications in lifespan studies[9], phenotyping and screening[10,11], nerve regeneration analyses[12], as well as for the investigation of worms’ behavioral dynamics[13] Such robust methods to study C. elegans embryos do not exist yet, whereas a microfluidic solution has been only proposed to study early embryo development for larger size model organisms, like Drosophila www.nature.com/scientificreports/. To enable systematic analysis of C. elegans embryonic morphogenesis, we developed a microfluidic platform for automated on-chip worm culture, creation of synchronized embryo arrays, and for long-term parallel live imaging at the single embryo level. Using our method to study a large number of embryos of different wild-type and mutant worm strains, we elucidated an outstanding issue regarding the role of UPRmt during early worm embryogenesis
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
