Lee Kyung Suk, Levine Erel
Department of Physics Education, Kongju National University; Department of Physics, Harvard University.
Department of Physics, Harvard University; FAS Center for Systems Biology, Harvard University;
J Vis Exp. 2018 May 2(135):57348. doi: 10.3791/57348.
In the last decade, microfluidic techniques have been applied to study small animals, including the nematode Caenorhabditis elegans, and have proved useful as a convenient live imaging platform providing capabilities for precise control of experimental conditions in real time. In this article, we demonstrate live imaging of individual worms employing WormSpa, a previously-published custom microfluidic device. In the device, multiple worms are individually confined to separate chambers, allowing multiplexed longitudinal surveillance of various biological processes. To illustrate the capability, we performed proof-of-principle experiments in which worms were infected in the device with pathogenic bacteria, and the dynamics of expression of immune response genes and egg laying were monitored continuously in individual animals. The simple design and operation of this device make it suitable for users with no previous experience with microfluidic-based experiments. We propose that this approach will be useful for many researchers interested in longitudinal observations of biological processes under well-defined conditions.
在过去十年中,微流控技术已被应用于研究包括线虫秀丽隐杆线虫在内的小型动物,并已被证明是一个方便的实时成像平台,能够实时精确控制实验条件。在本文中,我们展示了使用WormSpa(一种先前发表的定制微流控设备)对单个线虫进行实时成像。在该设备中,多条线虫被单独限制在不同的腔室中,从而能够对各种生物学过程进行多重纵向监测。为了说明其功能,我们进行了原理验证实验,在线虫感染致病细菌的情况下,对单个动物体内免疫反应基因的表达动态和产卵情况进行连续监测。该设备的简单设计和操作使其适用于以前没有微流控实验经验的用户。我们认为,这种方法对于许多有兴趣在明确条件下对生物学过程进行纵向观察的研究人员将是有用的。