Abstract
The nematode Caenorhabditis elegans is a suitable model organism in drug screening. Traditionally worms are grown on agar plates, posing many challenges for long-term culture and phenotyping of animals under identical conditions. Microfluidics allows for ‘personalized’ phenotyping, as microfluidic chips permit collecting individual responses over worms’ full life. Here, we present a multiplexed, high-throughput, high-resolution microfluidic approach to culture C. elegans from embryo to the adult stage at single animal resolution. We allocated single embryos to growth chambers, for observing the main embryonic and post-embryonic development stages and phenotypes, while exposing worms to up to 8 different well-controlled chemical conditions. Our approach allowed eliminating bacteria aggregation and biofilm formation-related clogging issues, which enabled us performing up to 80 hours of automated single worm culture studies. Our microfluidic platform is linked with an automated phenotyping code that registers organism-associated phenotypes at high-throughput. We validated our platform with a dose-response study of the anthelmintic drug tetramisole by studying its influence through the life cycle of the nematodes. In parallel, we could observe development effects and variations in single embryo and worm viability due to the bleaching procedure that is standardly used for harvesting the embryos from a worm culture agar plate.
Highlights
Www.nature.com/scientificreports microfluidic chips were combined with an agarose gel substrate, permitted single nematode manipulation and culture[17,18]
Microfluidic platforms for single worm culture starting from L121,22 or L423 stages were proposed, but they were operating at low-throughput and required an integration of on-chip components like pressure-activated valves that increased the complexity of the microfabrication
Another study demonstrated the possibility of life cycle culturing, starting from the embryo stage[28], but single-point entrances of growth chambers for such designs were susceptible to bacteria accumulation and biofilm formation and experimental parallelization was limited
Summary
Www.nature.com/scientificreports microfluidic chips were combined with an agarose gel substrate, permitted single nematode manipulation and culture[17,18]. Some microfluidic designs showed that single nematode loading in individual growth chambers was possible at higher throughput, but worm feeding was performed through single nozzle entrances[24,25,26,27]. Another study demonstrated the possibility of life cycle culturing, starting from the embryo stage[28], but single-point entrances of growth chambers for such designs were susceptible to bacteria accumulation and biofilm formation and experimental parallelization was limited. These devices still lacked the degree of automation needed to collect and analyze a significantly large amount of data, as required for high-throughput applications. Our platform simultaneously allowed studying the effects of a standard bleaching procedure on the health of embryos and worms
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