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线虫体内细菌转运的活体微流控研究。

An in vivo microfluidic study of bacterial transit in C. elegans nematodes.

机构信息

Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.

出版信息

Lab Chip. 2020 Aug 7;20(15):2696-2708. doi: 10.1039/d0lc00064g. Epub 2020 Jul 7.

Abstract

Caenorhabditis elegans (C. elegans) constitutes an important model organism for use in nutrition and aging studies. We report a novel method for studying the dynamics of Escherichia coli (E. coli) bacterial transit through the worms' intestine. A microfluidic chip was designed for alternating C. elegans on-chip culture and immobilization, thereby enabling periodic high-resolution time-lapse imaging at single-worm resolution over several days. Immobilization was achieved in a reversible way using arrays of tapered channels suitable for assay parallelization. Dedicated C. elegans feeding protocols were applied. Two E. coli bacterial strains, HT115 and OP50, respectively labeled with green fluorescent protein (GFP) and red fluorescent protein (RFP), were used as food source and imaged with fluorescence microscopy techniques to measure relevant parameters of the bacterial transit process. Feeding behavior and E. coli transit dynamics in the whole intestinal tract of the worms were characterized in an automated way over the first 3 days of adulthood, revealing both fast transit phenomena and variations in microbial accumulation. In particular, we studied the bacterial food transit periodicity in wild-type and eat-2 (ad465) mutant C. elegans strains in both trapped and free-swimming conditions. In order to further demonstrate the versatility of our microfluidic platform, we also studied drug-induced modifications of the bacterial transit by measuring the response of the worms' intestine to exposure to the neurotransmitter serotonin.

摘要

秀丽隐杆线虫(C. elegans)是营养和衰老研究中重要的模式生物。我们报告了一种研究大肠杆菌(E. coli)细菌穿过线虫肠道的动力学的新方法。设计了一种微流控芯片,用于交替进行线虫的片上培养和固定,从而能够在数天内以单只线虫的分辨率进行周期性的高分辨率时程成像。通过适用于分析并行化的锥形通道阵列以可逆的方式实现固定。应用了专门的秀丽隐杆线虫饲养方案。使用分别标记有绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)的两种大肠杆菌菌株 HT115 和 OP50 作为食物源,并通过荧光显微镜技术进行成像,以测量细菌转运过程的相关参数。在成虫期的前 3 天,以自动方式对整个肠道中的线虫摄食行为和 E. coli 转运动力学进行了表征,揭示了快速转运现象和微生物积累的变化。特别是,我们在被困和自由游动条件下研究了野生型和 eat-2(ad465)突变体秀丽隐杆线虫菌株中细菌食物转运的周期性。为了进一步证明我们的微流控平台的多功能性,我们还通过测量蠕虫肠道对神经递质血清素暴露的反应来研究药物诱导的细菌转运修饰。

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