A microfluidic device and automatic counting system for the study of C. elegans reproductive aging.-猫眼课题宝

DOI：

10.1039/c4lc01028k

发表时间：

2015-01-21

期刊：

Lab on a chip

影响因子：

6.1

通讯作者：

中科院分区：

工程技术1区

文献类型：

作者：

Li S;Stone HA;Murphy CT

来源链接：

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4277648/?report=classic

线虫秀丽线虫(Cinehabditis elegans，C.elegans)是研究生殖衰老的理想模型，因为它的寿命短，在成年中期停止繁殖，而且调节寿命的途径具有很强的保守性。在其一生中，野生线虫两性通常产卵约200-300个自受精的可孵化的卵，主要发生在成虫的前三到五天。在这里，我们报告了一种微流控分析和实时、自动后代计数系统的发展，该系统记录了许多线虫两性个体的后代计数信息。与传统的平板分析方法相比，该系统具有许多优点。非增殖性细菌的流动不仅喂养蠕虫，还通过将母亲和后代分开的过滤器将刚刚孵化的幼虫冲走。用一种新的算法检测和记录被冲出密室的子代。在我们目前的设计中，一个设备包含多达16个单独的腔室。在这里，我们展示了野生型(N2)和daf-2(胰岛素受体)突变体的实时后代生产信息的例子。我们相信，该系统有可能成为一种强大的、高时间分辨率的工具来研究线虫的详细繁殖。

The nematode Caenorhabditis elegans (C. elegans) is an excellent model to study reproductive aging because of its short life span, its cessation of reproduction in mid-adulthood, and the strong conservation of pathways that regulate longevity. During its lifetime, a wild-type C. elegans hermaphrodite usually lays about 200–300 self-fertilized hatchable eggs, which mainly occurs in the first three to five days of adulthood. Here, we report the development of a microfluidic assay and a real-time, automatic progeny counting system that records progeny counting information from many individual C. elegans hermaphrodites. This system offers many advantages compared to conventional plate assays. The flow of non-proliferating bacteria not only feeds the worms but also flushes the just-hatched young progeny through a filter that separates mothers from their offspring. The progeny that are flushed out of the chamber are detected and recorded using a novel algorithm. In our current design, one device contains as many as 16 individual chambers. Here we show examples of real-time progeny production information from wild-type (N2) and daf-2 (insulin receptor) mutants. We believe that this system has the potential to become a powerful, high time-resolution tool to study the detailed reproduction of C. elegans.