Department of Physics, University of Ottawa, Ottawa, Ontario, Canada.
Ottawa Institute of Systems Biology, Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.
Sci Rep. 2018 Dec 13;8(1):17809. doi: 10.1038/s41598-018-36000-3.
An imaging-integrated microfluidic cell volume sensor was used to evaluate the volumetric growth rate of single cells from a Saccharomyces cerevisiae population exhibiting two phenotypic expression states of the PDR5 gene. This gene grants multidrug resistance by transcribing a membrane transporter capable of pumping out cytotoxic compounds from the cell. Utilizing fluorescent markers, single cells were isolated and trapped, then their growth rates were measured in two on-chip environments: rich media and media dosed with the antibiotic cycloheximide. Approximating growth rates to first-order, we assessed the fitness of individual cells and found that those with low PDR5 expression had higher fitness in rich media whereas cells with high PDR5 expression had higher fitness in the presence of the drug. Moreover, the drug dramatically reduced the fitness of cells with low PDR5 expression but had comparatively minimal impact on the fitness of cells with high PDR5 expression. Our experiments show the utility of this imaging-integrated microfluidic cell volume sensor for high-resolution, single-cell analysis, as well as its potential application for studies that characterize and compare the fitness and morphology of individual cells from heterogeneous populations under different growth conditions.
使用成像集成微流控细胞体积传感器来评估表现出 PDR5 基因两种表型表达状态的酿酒酵母群体中单细胞的体积增长率。该基因通过转录一种能够将细胞毒性化合物从细胞中泵出的膜转运蛋白赋予多药耐药性。利用荧光标记物,分离并捕获单个细胞,然后在两种芯片环境中测量它们的生长速率:富培养基和用抗生素环己酰亚胺处理的培养基。将生长速率近似为一级,我们评估了单个细胞的适应性,发现低 PDR5 表达的细胞在富培养基中适应性更高,而高 PDR5 表达的细胞在存在药物时适应性更高。此外,该药物大大降低了低 PDR5 表达细胞的适应性,但对高 PDR5 表达细胞的适应性影响相对较小。我们的实验表明,这种成像集成微流控细胞体积传感器可用于高分辨率单细胞分析,以及用于研究在不同生长条件下从异质群体中表征和比较单个细胞的适应性和形态的潜在应用。