Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy
Quantifying the forces guiding microbial cell adhesion using single-cell force spectroscopy
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DOI：
10.1038/nprot.2014.066
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发表时间：
2014-05-01
期刊：
影响因子：
14.8
通讯作者：
Dufrene, Yves F.
中科院分区：
文献类型：
--
作者：
Beaussart, Audrey;El-Kirat-Chatel, Sofiane;Dufrene, Yves F.
在过去的几十年里，已经发展了几种方法（如电子显微镜、流室实验、表面化学分析、表面电荷和表面疏水性测量）来研究控制微生物细胞与其他细胞和各种其他底物粘附的机制。然而，没有一种传统的方法能够在单细胞水平上观察粘附力。近年来，原子力显微镜（AFM）已被用于测量驱动微生物在单细胞基础上的粘附力。该方法被称为单细胞力谱（SCSFS），包括将单个活细胞固定在AFM悬臂上，并测量细胞探针与固体衬底或另一个细胞之间的相互作用力。在这里，我们提出了SCFS方案，我们已经开发了用于量化医学上重要微生物的细胞粘附力。虽然我们主要关注益生菌植物乳杆菌，但我们也表明我们的程序适用于病原体，如表皮葡萄球菌和白色念珠菌。对于训练有素的显微镜师，整个程序可以在1周内掌握。
During the past decades, several methods (e. g., electron microscopy, flow chamber experiments, surface chemical analysis, surface charge and surface hydrophobicity measurements) have been developed to investigate the mechanisms controlling the adhesion of microbial cells to other cells and to various other substrates. However, none of the traditional approaches are capable of looking at adhesion forces at the single-cell level. In recent years, atomic force microscopy (AFM) has been instrumental in measuring the forces driving microbial adhesion on a single-cell basis. The method, known as single-cell force spectroscopy (SCSFS), consists of immobilizing a single living cell on an AFM cantilever and measuring the interaction forces between the cellular probe and a solid substrate or another cell. Here we present SCFS protocols that we have developed for quantifying the cell adhesion forces of medically important microbes. Although we focus mainly on the probiotic bacterium Lactobacillus plantarum, we also show that our procedures are applicable to pathogens, such as the bacterium Staphylococcus epidermidis and the yeast Candida albicans. For well-trained microscopists, the entire protocol can be mastered in 1 week.