School of Chemical & Biomolecular Engineering, Georgia Institute of Technology, 311 Ferst Dr, NW, Atlanta, GA, 30332-0100, USA.
Interdisciplinary Program of Bioengineering, Georgia Institute of Technology, 311 Ferst Dr, NW, Atlanta, GA, 30332-0100, USA.
Lab Chip. 2011 Nov 7;11(21):3689-3697. doi: 10.1039/c1lc20400a. Epub 2011 Sep 20.
The nematode Caenorhabditis elegans is an important model organism in genetic research and drug screening because of its relative simplicity, ease of maintenance, amenability to simple genetic manipulation, and relevance to human biology. However, their small size and mobility make nematodes difficult to physically manipulate, particularly with spatial and temporal precision. We have developed a microfluidic device to overcome these challenges and enable fast behavior-based chemical screening in C. elegans. The key components of this easy-to-use device allow rapid loading and housing of C. elegans in a chamber array for chemical screening. A simple two-step loading process enables simultaneous loading of a large number of animals within a few minutes without using any expensive/active off-chip components. In addition, chemicals can be precisely delivered to the worms and exchanged with high temporal precision. To demonstrate this feature and the ability to measure time dependent responses to chemicals, we characterize the transient response of worms exposed to different concentrations of anesthetics. We then use the device to study the effect of chemical signals from hermaphrodite worms on male behavior. The ability of the device to maintain a large number of free moving animals in one field of view over a long period of time permits us to demonstrate an increase in the incidence of a specific behavior in males subjected to worm-conditioned medium. Because our device allows monitoring of a large number of worms with single-animal resolution, we envision that this platform will greatly expedite chemical screening in C. elegans.
秀丽隐杆线虫是遗传研究和药物筛选的重要模式生物，因为它相对简单、易于维持、易于进行简单的遗传操作，并且与人类生物学相关。然而，它们的体积小且移动性强，使得线虫难以进行物理操作，特别是在空间和时间精度方面。我们开发了一种微流控设备来克服这些挑战，实现秀丽隐杆线虫的快速基于行为的化学筛选。该易用型设备的关键组件允许快速加载和容纳线虫在腔室阵列中进行化学筛选。简单的两步加载过程能够在几分钟内同时加载大量动物，而无需使用任何昂贵/有源的片外组件。此外，化学物质可以精确地输送到蠕虫体内，并以高精度进行交换。为了展示这一功能和测量对化学物质的时间依赖性反应的能力，我们对暴露于不同浓度麻醉剂的蠕虫的瞬态反应进行了表征。然后，我们使用该设备研究了雌雄同体蠕虫释放的化学信号对雄性行为的影响。该设备能够在一个视场中长时间维持大量自由移动的动物的能力，使我们能够证明在受到蠕虫条件培养基处理的雄性中，特定行为的发生率增加。因为我们的设备允许以单只动物分辨率监测大量蠕虫，所以我们设想该平台将极大地加快秀丽隐杆线虫的化学筛选速度。
