1Centre of Microbial and Plant Genetics, KU Leuven-University of Leuven, Leuven, 3001 Belgium.
VIB-KU Leuven Center of Microbiology, Leuven, 3001 Belgium.
Commun Biol. 2019 Jul 23;2:269. doi: 10.1038/s42003-019-0480-9. eCollection 2019.
Image-based high-throughput screening strategies for quantifying morphological phenotypes have proven widely successful. Here we describe a combined experimental and multivariate image analysis approach for systematic large-scale phenotyping of morphological dynamics in bacteria. Using off-the-shelf components and software, we established a workflow for high-throughput time-resolved microscopy. We then screened the single-gene deletion collection of for antibiotic-induced morphological changes. Using single-cell quantitative descriptors and supervised classification methods, we measured how different cell morphologies developed over time for all strains in response to the β-lactam antibiotic cefsulodin. 191 strains exhibit significant variations under antibiotic treatment. Phenotypic clustering provided insights into processes that alter the antibiotic response. Mutants with stable bulges show delayed lysis, contributing to antibiotic tolerance. Lipopolysaccharides play a crucial role in bulge stability. This study demonstrates how multiparametric phenotyping by high-throughput time-resolved imaging and computer-aided cell classification can be used for comprehensively studying dynamic morphological transitions in bacteria.
基于图像的高通量筛选策略在定量形态表型方面已被证明非常成功。在这里，我们描述了一种结合实验和多元图像分析的方法，用于对细菌形态动力学进行系统的大规模表型分析。我们使用现成的组件和软件，建立了高通量时分辨微镜的工作流程。然后，我们对 的单基因缺失文库进行了抗生素诱导形态变化的筛选。使用单细胞定量描述符和监督分类方法，我们测量了所有菌株在β-内酰胺类抗生素头孢菌素处理下，随时间推移的不同细胞形态如何发展。191 株在抗生素处理下表现出显著的变化。表型聚类提供了关于改变抗生素反应的过程的见解。具有稳定隆起的突变体显示出延迟裂解，导致抗生素耐药性。脂多糖在隆起稳定性中起着至关重要的作用。这项研究展示了如何通过高通量时分辨成像和计算机辅助细胞分类的多参数表型分析来全面研究细菌中的动态形态转变。
