Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, Minneapolis, MN 55455, USA.
Lab Chip. 2013 Mar 7;13(5):947-54. doi: 10.1039/c2lc41196b. Epub 2013 Jan 18.
We designed a microfluidic chemostat consisting of 600 sub-micron trapping/growth channels connected to two feeding channels. The microchemostat traps E. coli cells and forces them to grow in lines for over 50 generations. Excess cells, including the mother cells captured at the start of the process, are removed from both ends of the growth channels by the media flow. With the aid of time-lapse microscopy, we have monitored dynamic properties such as growth rate and GFP expression at the single-cell level for many generations while maintaining a population of bacteria of similar age. We also use the microchemostat to show how the population responds to dynamic changes in the environment. Since more than 100 individual bacterial cells are aligned and immobilized in a single field of view, the microchemostat is an ideal platform for high-throughput intracellular measurements. We demonstrate this capability by tracking with sub-diffraction resolution the movements of fluorescently tagged loci in more than one thousand cells on a single device. The device yields results comparable to conventional agar microscopy experiments with substantial increases in throughput and ease of analysis.
我们设计了一种由 600 个亚微米捕获/生长通道组成的微流控恒化器，这些通道连接到两个进料通道上。微恒化器捕获大肠杆菌细胞，并迫使它们在线生长超过 50 代。通过培养基流动，从生长通道的两端除去多余的细胞，包括在该过程开始时捕获的母细胞。借助延时显微镜，我们在保持类似年龄的细菌种群的同时，在单细胞水平上监测了许多代的动态特性，如生长速率和 GFP 表达。我们还使用微恒化器来展示种群如何响应环境中的动态变化。由于超过 100 个单个细菌细胞在单个视场中对齐和固定，因此微恒化器是高通量细胞内测量的理想平台。我们通过以亚衍射分辨率跟踪荧光标记的基因座在单个装置上的 1000 多个细胞中的运动，证明了这种能力。该装置的结果与传统琼脂显微镜实验相当，具有更高的通量和更易于分析的优势。
