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Long-term time-lapse microscopy of C. elegans post-embryonic development

机译:秀丽隐杆线虫胚胎发育的长期延时显微镜

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摘要

We present a microscopy technique that enables long-term time-lapse microscopy at single-cell resolution in moving and feeding Caenorhabditis elegans larvae. Time-lapse microscopy of C. elegans post-embryonic development is challenging, as larvae are highly motile. Moreover, immobilization generally leads to rapid developmental arrest. Instead, we confine larval movement to microchambers that contain bacteria as food, and use fast image acquisition and image analysis to follow the dynamics of cells inside individual larvae, as they move within each microchamber. This allows us to perform fluorescence microscopy of 10–20 animals in parallel with 20 min time resolution. We demonstrate the power of our approach by analysing the dynamics of cell division, cell migration and gene expression over the full ∼48 h of development from larva to adult. Our approach now makes it possible to study the behaviour of individual cells inside the body of a feeding and growing animal.
机译:我们提出了一种显微技术,该技术能够在移动和喂养秀丽隐杆线虫幼虫的单细胞分辨率下进行长期延时显微镜操作。线虫的胚胎发育后的延时显微镜具有挑战性,因为幼虫具有很高的运动能力。而且,固定化通常导致快速的发育停滞。取而代之的是,我们将幼虫的活动限制在含有细菌作为食物的微腔室中,并使用快速图像采集和图像分析来跟踪各个幼体中每个幼体中细胞移动的动态。这使我们能够以20µmin的时间分辨率对10–20只动物进行荧光显微镜检查。我们通过分析从幼虫到成虫的整个〜48 h内的细胞分裂,细胞迁移和基因表达的动力学,证明了我们方法的力量。现在,我们的方法使研究饲养和正在生长的动物体内的单个细胞的行为成为可能。

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