CeMM-WormTracker: Long-term, single-worm level tracking and phenotyping of C. elegans cultivated in CeMM on an integrated and enclosed microfluidic device

IF 3.3 3区 化学 Q2 CHEMISTRY, ANALYTICAL
Analyst Pub Date : 2025-07-11 DOI:10.1039/d5an00394f
Qianqian Yang, Runtao Zhong, Mengyu Wang, Wenbo Chang, Kexin Chen, Yeqing Sun
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Abstract

Caenorhabditis elegans (C. elegans) is a typical model organism that has predominantly relied on growth using a bacterial diet, presenting limitations for automated experimentations (mainly due to the requirement of periodic transfer to new plates), and accuracy of results (because of possible interference by bacterial metabolism in liquid media), which makes flexible manipulation and long-term tracking difficult. C. elegans Maintenance Medium (CeMM), a chemically defined sterile liquid medium, holds the potential to solve these problems. Population-level studies of C. elegans cultured in CeMM have shown that nematode development slows, fecundity declines, lifespan increases, lipid and protein stores decrease, and gene expression changes relative to that on a bacterial diet. However, automated cultivation of C. elegans in CeMM, long-term tracking and phenotyping at single-worm level remains challenging. Here, we developed a chamber-array chip (WormChip-1.8) and an integrated and enclosed microfluidic device, CeMM-WormTracker, with the capability of automating fluid control and worm manipulation for single-worm level tracking and phenotyping of C. elegans grown in CeMM. By using a microscope, the CeMM-WormTracker allows for observation of C. elegans development, motility, reproduction, and survival for a long period of time. Our data demonstrate that it is possible to longitudinally track and phenotype the growth and development of nematodes at single-worm level for at least 75 days within the device, enabling comprehensive monitoring of the whole reproductive period and lifespan in sterile liquid culture, which is difficult for well plate-based experiments. Comparing with the results from 96-well plates, the development, activity, and reproduction of nematodes in the microfluidic device seems more stable. Thus, the CeMM-WormTracker provides feasible solutions for automated and high-throughput experimentations in studies of C. elegans, both on the ground and in orbit.
CeMM- wormtracker:在集成和封闭的微流体装置上对在CeMM中培养的秀丽隐杆线虫进行长期、单虫水平的跟踪和表型分析
秀丽隐杆线虫(C. elegans)是一种典型的模式生物,主要依靠细菌饮食生长,存在自动化实验的局限性(主要是因为需要定期转移到新的培养皿),结果的准确性(因为可能受到液体介质中细菌代谢的干扰),这使得灵活操作和长期跟踪变得困难。秀丽隐杆线虫维持培养基(CeMM)是一种化学定义的无菌液体培养基,具有解决这些问题的潜力。在CeMM中培养的秀丽隐杆线虫的种群水平研究表明,与细菌饮食相比,线虫发育减慢,繁殖力下降,寿命延长,脂质和蛋白质储存减少,基因表达发生变化。然而,秀丽隐杆线虫在CeMM中的自动培养、单虫水平的长期跟踪和表型分析仍然具有挑战性。在此,我们开发了一种室阵列芯片(WormChip-1.8)和一个集成和封闭的微流体装置(CeMM- wormtracker),具有自动流体控制和蠕虫操作的能力,用于在CeMM中生长的线虫的单蠕虫水平跟踪和表型分析。通过使用显微镜,CeMM-WormTracker可以长时间观察秀丽隐杆线虫的发育、运动、繁殖和存活。我们的数据表明,在该装置内,可以在单个线虫水平上纵向跟踪和表型线虫的生长和发育至少75天,从而可以在无菌液体培养中全面监测整个生殖期和寿命,这在孔板实验中是困难的。与96孔板实验结果相比,微流控装置中线虫的发育、活动和繁殖更加稳定。因此,CeMM-WormTracker为地面和轨道上秀丽隐杆线虫研究的自动化和高通量实验提供了可行的解决方案。
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来源期刊
Analyst
Analyst 化学-分析化学
CiteScore
7.80
自引率
4.80%
发文量
636
审稿时长
1.9 months
期刊介绍: "Analyst" journal is the home of premier fundamental discoveries, inventions and applications in the analytical and bioanalytical sciences.
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