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基于导向电趋性行为的高效秀丽隐杆线虫发育阶段同步微流控分选装置。

Highly efficient microfluidic sorting device for synchronizing developmental stages of C. elegans based on deflecting electrotaxis.

机构信息

Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Systems Biology Theme, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China.

出版信息

Lab Chip. 2015 Jun 7;15(11):2513-21. doi: 10.1039/c5lc00354g. Epub 2015 May 12.

Abstract

C. elegans as a powerful model organism has been widely used in fundamental biological studies. Many of these studies frequently need a large number of different stage-synchronized worms due to the stage-specific features of C. elegans among 4 distinct larval stages and the adult stage. In this work, we present an interesting and cost-effective microfluidic approach to realize simultaneous sorting of C. elegans of different developmental stages by deflecting electrotaxis. The microfluidic device was fabricated using PDMS consisting of symmetric sorting channels with specific angles, which was further hybridized to an agarose plate. While applying an electric field, different stages of C. elegans would crawl to the negative pore with different angles due to their deflecting electrotaxis. Thus, the worms were separated and synchronized by stages. lon-2 mutant was further used to study this electrotactic response and the results indicated that the body size plays a key role in determining the deflecting angle in matured adult worms. In addition to discriminating wild-type hermaphrodites, it could also be employed to sort mutants with abnormal development sizes and males. Therefore, our device provided a versatile and highly efficient platform for sorting C. elegans to meet the requirement of large numbers of different stage-synchronized worms. It can also be further used to investigate the neuronal basis of deflecting electrotaxis in worms.

摘要

秀丽隐杆线虫作为一种强大的模式生物,已被广泛应用于基础生物学研究。由于秀丽隐杆线虫有 4 个不同的幼虫阶段和成虫阶段,具有阶段特异性特征,因此许多这些研究经常需要大量不同阶段同步的线虫。在这项工作中,我们提出了一种有趣且具有成本效益的微流控方法,通过转向电泳实现对不同发育阶段秀丽隐杆线虫的同时分选。该微流控装置由 PDMS 制成,包含具有特定角度的对称分选通道,进一步与琼脂糖板杂交。当施加电场时,由于转向电泳,不同阶段的秀丽隐杆线虫会因角度不同而爬行到负孔。因此,线虫通过阶段进行分离和同步。进一步使用 lon-2 突变体来研究这种趋电反应,结果表明,在成熟的成虫中,体型大小在确定转向角度方面起着关键作用。除了区分野生型雌雄同体外,它还可用于分选发育大小异常的突变体和雄性。因此,我们的设备为分选秀丽隐杆线虫提供了一个通用且高效的平台,以满足大量不同阶段同步线虫的需求。它还可以进一步用于研究线虫转向电泳的神经元基础。

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