PDMS filter structures for size-dependent larval sorting and on-chip egg extraction of C. elegans.,Lab on a Chip

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PDMS filter structures for size-dependent larval sorting and on-chip egg extraction of C. elegans.
Lab on a Chip ( IF 6.1 ) Pub Date : 2019-12-03 , DOI: 10.1039/c9lc00949c
Huseyin Baris Atakan ₁ , Furkan Ayhan ₁ , Martin A M Gijs ₁

Affiliation

C. elegans-based assays require age-synchronized populations prior to experimentation to achieve standardized sets of worm populations, due to which age-induced heterogeneous phenotyping effects can be avoided. There have been several approaches to synchronize populations of C. elegans at certain larval stages; however, many of these methods are tedious, complex and have low throughput. In this work, we demonstrate a polydimethylsiloxane (PDMS) microfluidic filtering device for high-throughput, efficient, and extremely rapid sorting of mixed larval populations of C. elegans. Our device consists of three plasma-activated and bonded PDMS parts and permits sorting of mixed populations of two consecutive larval stages in a matter of minutes. After sorting, we also retain the remaining larval stage of the initially mixed worm population on the chip, thereby enabling collection of the two sorted larval populations from the device. We demonstrated that the target larvae could be collected from a mixed worm population by cascading these devices. Our approach is based on only passive hydrodynamics filter structures, resulting in a user-friendly and reusable tool. In addition, we employed the equivalent of a standard bleaching procedure that is practiced in standard worm culture on agar plates for embryo harvesting on our chip, and we demonstrated rapid egg extraction and subsequent harvesting of a synchronized L1 larvae population.

中文翻译：

PDMS过滤器结构，用于秀丽隐杆线虫大小相关的幼体分选和片上卵提取。

基于秀丽隐杆线虫的测定法在进行实验之前需要年龄同步的种群，以实现标准化的蠕虫种群，从而可以避免年龄引起的异质表型效应。有几种方法可以使某些幼虫阶段的秀丽隐杆线虫种群同步。然而，这些方法中的许多繁琐，复杂且吞吐量低。在这项工作中，我们演示了聚二甲基硅氧烷（PDMS）微流体过滤装置，可用于对秀丽隐杆线虫的混合幼虫种群进行高通量，高效和极快速的分类。我们的设备由三个等离子体激活和键合的PDMS部件组成，并允许在数分钟内对两个连续幼体阶段的混合种群进行分选。排序后，我们还将芯片上最初混合蠕虫种群的剩余幼体阶段保留在芯片上，从而可以从设备中收集两个分类的幼虫种群。我们证明，通过级联这些设备，可以从混合蠕虫种群中收集目标幼虫。我们的方法仅基于被动流体力学滤清器结构，因此是一种用户友好且可重复使用的工具。此外，我们采用了与标准漂白程序等效的方法，该程序在琼脂平板上的标准蠕虫培养中进行，以在我们的芯片上收获胚胎，并且我们证明了快速的卵提取和随后的同步L1幼虫种群的收获。从而产生了一种用户友好且可重复使用的工具。此外，我们采用了与标准漂白程序等效的方法，该程序在琼脂平板上的标准蠕虫培养中进行，以在我们的芯片上收获胚胎，并且我们证明了快速的卵提取和随后的同步L1幼虫种群的收获。从而产生了一种用户友好且可重复使用的工具。此外，我们采用了与标准漂白程序等效的方法，该程序在琼脂平板上的标准蠕虫培养中进行，以在我们的芯片上收获胚胎，并且我们证明了快速的卵提取和随后的同步L1幼虫种群的收获。

更新日期：2019-12-18

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