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An automated microfluidic platform for C. elegans embryo arraying phenotypingand long-term live imaging

机译:用于秀丽隐杆线虫胚胎排列表型分析和长期实时成像

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摘要

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.
机译:对胚胎发育实时动态的研究需要一种柔和的胚胎处理方法,在整个胚胎发生过程中进行长期实时成像的可能性以及并行化以提供种群统计数据,同时保持单个胚胎的分辨率。我们描述了一种自动化方法,可以完全满足秀丽隐杆线虫的胚胎的这些要求,秀丽隐杆线虫是生物医学研究中使用最多的模型生物之一。我们开发了一种微流体平台,该平台利用纯被动流体力学运行芯片上的蠕虫培养,从中获得同步的胚胎种群,并将这些胚胎固定在培养箱微阵列中,以进行长期的高分辨率光学成像。我们成功地利用我们的平台研究了整个胚胎发育过程中的形态发生和线粒体生物发生,并阐明了线虫未折叠蛋白应答(UPR mt )在秀丽隐杆线虫胚胎发生中的作用。我们的方法通常可用于胚胎水平的蛋白质表达和发育研究,但也可为了解衰老过程以及特别是与年龄相关的疾病提供线索。

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