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A high-throughput device for size based separation of C. elegans developmental stages

机译:一种高通量的设备用于基于尺寸的秀丽隐杆线虫发育阶段分离

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摘要

>Caenorhabditis elegans is a widely used model organism to study development, aging and behavior. Many of these biological studies require staging a large number of worms to assay a synchronized population of animals. Conventional synchronization techniques such as manual picking, gravity stratification and chemical bleaching are labor-intensive and could perturb animals’ physiology. Thus, there is a need for a simple inexpensive technology to sort a mixed population of worms based on their developmental stages with minimal perturbation. Here we demonstrate a simple but accurate and high-throughput technique to sort based on animal size, which correlates well with developmental stages. The device consists of an array of geometrically optimized pillars that act as a sieve to allow worms of specific sizes to rapidly move through. With optimized chamber heights, pillar spacing and driving pressures, these binary separation devices are capable of independently separating a mixture of worms at two different stages at average efficiency of around 95%, and throughput of hundreds of worms per minute. In addition, when four devices are used sequentially, we demonstrate the ability to stratify a mixture of worms of all developmental stages with >85% overall efficiency.
机译:>秀丽隐杆线虫是一种广泛用于研究发育,衰老和行为的模型生物。这些生物学研究中的许多都需要分阶段大量蠕虫来分析动物的同步种群。常规的同步技术,例如人工采摘,重力分层和化学漂白,是劳动密集型的,可能会扰乱动物的生理。因此,需要一种简单廉价的技术来根据蠕虫的发育阶段以最小的扰动对蠕虫的混合种群进行分类。在这里,我们演示了一种简单但准确且高通量的技术,可以根据动物的大小进行分类,这与发育阶段密切相关。该设备由一系列几何优化的支柱组成,这些支柱充当筛子,以允许特定大小的蠕虫快速通过。通过优化的腔室高度,支柱间距和驱动压力,这些二元分离设备能够在两个不同的阶段独立分离蠕虫的混合物,平均效率约为95%,每分钟可吞噬数百个蠕虫。此外,当依次使用四个设备时,我们证明了对所有发育阶段的蠕虫混合物进行分层的能力,总体效率> 85%。

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