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Automated Platform for Long-Term Culture and High-Content Phenotyping of Single C. elegans Worms

机译:线虫蠕虫的长期培养和高内涵表型自动化平台

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摘要

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.
机译:线虫秀丽隐杆线虫是药物筛选中合适的模型生物。传统上,蠕虫生长在琼脂平板上,在相同条件下对动物的长期培养和表型构成许多挑战。微流体技术可以进行“个性化”表型分析,因为微流体芯片可以收集蠕虫整个生命周期内的各个响应。在这里,我们提出了一种以单一动物分辨率从种子到成年阶段培养线虫的多重,高通量,高分辨率微流控方法。我们将单个胚胎分配到生长室,以观察主要的胚胎和胚后发育阶段和表型,同时将蠕虫暴露于多达8种不同的良好控制的化学条件下。我们的方法可以消除细菌聚集和与生物膜形成有关的堵塞问题,这使我们能够进行长达80个小时的自动单蠕虫培养研究。我们的微流体平台与自动表型代码链接,该代码以高通量注册与生物相关的表型。我们通过研究其在线虫生命周期中的影响,对驱虫药四咪唑进行了剂量反应研究,从而验证了我们的平台。同时,由于漂洗程序通常用于从蠕虫培养琼脂板上收获胚胎,因此我们可以观察到发育效果以及单个胚胎和蠕虫生存力的变化。

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