Lee Kyung Suk, Lee Lucy E, Levine Erel
Department of Physics and Center for Systems Biology, Harvard University, Cambridge, MA 02138, USA.
Sci Rep. 2016 Oct 24;6:35862. doi: 10.1038/srep35862.
Small animals such as the roundworm C. elegans are excellent models for studying bacterial infection and host response, as well as for genetic and chemical screens. A key methodology is the killing assay, in which the number of surviving animals is tracked as a function of the time post infection. This is a labor-intensive procedure, prone to human error and subjective choices, and often involves undesired perturbation to the animals and their environment. In addition, the survival of animals is just one aspect of a multi-dimensional complex biological process. Here we report a microfluidic-based approach for performing killing assays in worms, compatible with standard assays performed on solid media. In addition to providing accurate and reproducible survival curves at a considerably reduced labor, this approach allows acquisition of a multitude of quantitative data with minimal undesired perturbations. These measurements are obtained automatically at a worm-by-worm resolution using a custom image processing workflow. The proposed approach is simple, scalable, and extendable, and is significantly more economical than standard manual protocols.
诸如秀丽隐杆线虫之类的小动物是研究细菌感染与宿主反应以及进行基因和化学筛选的优秀模型。一种关键方法是杀伤试验，即追踪感染后存活动物的数量随时间的变化。这是一个劳动密集型过程，容易出现人为误差和主观选择，并且常常会对动物及其环境造成不必要的干扰。此外，动物的存活只是一个多维复杂生物过程的一个方面。在此，我们报告一种基于微流控的方法，用于在线虫中进行杀伤试验，该方法与在固体培养基上进行的标准试验兼容。除了以大大减少的工作量提供准确且可重复的存活曲线外，这种方法还能够以最小的不必要干扰获取大量定量数据。这些测量是使用定制的图像处理工作流程以逐个线虫的分辨率自动获得的。所提出的方法简单、可扩展且可延伸，并且比标准的手动方案经济得多。
