Caenorhabditis elegans embryos have been widely used to study cellular processes and developmental regulation at early stages.However,most existing microfluidic devices focus on the studies of larval or adult worms ra...Caenorhabditis elegans embryos have been widely used to study cellular processes and developmental regulation at early stages.However,most existing microfluidic devices focus on the studies of larval or adult worms rather than embryos.To accurately study the real-time dynamics of embryonic development under different conditions,many technical barriers must be overcome;these can include single-embryo sorting and immobilization,precise control of the experimental environment,and long-term live imaging of embryos.This paper reports a spiral microfluidic device for effective sorting,trapping,and long-term live imaging of single C.elegans embryos under precisely controlled experimental conditions.The device successfully sorts embryos from a mixed population of C.elegans at different developmental stages via Dean vortices generated inside a spiral microchannel and traps the sorted embryos at single-cell resolution through hydrodynamic traps on the sidewall of the spiral channel for long-term imaging.Through the well-controlled microenvironment inside the microfluidic device,the response of the trapped C.elegans embryos to mechanical and chemical stimulation can be quantitatively measured.The experimental results show that a gentle hydrodynamic force would induce faster growth of embryos,and embryos developmentally arrested in the high-salinity solution could be rescued by the M9 buffer.The microfluidic device provides new avenues for easy,rapid,high-content screening of C.elegans embryos.展开更多
基金supported by the Natural Sciences and Engineering Research Council of Canada (grant numbers:RGPIN-2017-06374,RGPAS-2017-507980,and RGPIN-2022-05039)the Canadian Institutes of Health Research (grant number:PJT-180365)+4 种基金the Canada Foundation for Innovation (grant number:JELF-38428)support from the National Natural Science Foundation of China (62273247)the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (20KJA460008)to C.Ru is acknowledgedsupport from the Natural Science Foundation of the Jiangsu Higher Education Institutions of China (20KJB460024)the Young Scholar Program of Jiangsu Science and Technology (BK2020041995).
文摘Caenorhabditis elegans embryos have been widely used to study cellular processes and developmental regulation at early stages.However,most existing microfluidic devices focus on the studies of larval or adult worms rather than embryos.To accurately study the real-time dynamics of embryonic development under different conditions,many technical barriers must be overcome;these can include single-embryo sorting and immobilization,precise control of the experimental environment,and long-term live imaging of embryos.This paper reports a spiral microfluidic device for effective sorting,trapping,and long-term live imaging of single C.elegans embryos under precisely controlled experimental conditions.The device successfully sorts embryos from a mixed population of C.elegans at different developmental stages via Dean vortices generated inside a spiral microchannel and traps the sorted embryos at single-cell resolution through hydrodynamic traps on the sidewall of the spiral channel for long-term imaging.Through the well-controlled microenvironment inside the microfluidic device,the response of the trapped C.elegans embryos to mechanical and chemical stimulation can be quantitatively measured.The experimental results show that a gentle hydrodynamic force would induce faster growth of embryos,and embryos developmentally arrested in the high-salinity solution could be rescued by the M9 buffer.The microfluidic device provides new avenues for easy,rapid,high-content screening of C.elegans embryos.
