Cellular Microbiology and Physics of Infection Group-Lille Centre for Infection and Immunity, CNRS UMR8204, INSERM U1019, Lille University Hospital Centre, University of Lille, Institut Pasteur de Lille, Lille, France.
Bruker Nano Surfaces Division, Santa Barbara, California, United States of America.
PLoS One. 2019 Mar 15;14(3):e0213853. doi: 10.1371/journal.pone.0213853. eCollection 2019.
In the last 20 years, atomic force microscopy (AFM) has emerged as a ubiquitous technique in biological research, allowing the analysis of biological samples under near-physiological conditions from single molecules to living cells. Despite its growing use, the low process throughput remains a major drawback. Here, we propose a solution validated on a device allowing a fully automated, multi-sample analysis. Our approach is mainly designed to study samples in fluid and biological cells. As a proof of concept, we demonstrate its feasibility applied to detect and scan both fixed and living bacteria before completion of data processing. The effect of two distinct treatments (i.e. gentamicin and heating) is then evidenced on physical parameters of fixed Yersinia pseudotuberculosis bacteria. The multi-sample analysis presented allows an increase in the number of scanned samples while limiting the user's input. Importantly, cantilever cleaning and control steps are performed regularly-as part of the automated process-to ensure consistent scanning quality. We discuss how such an approach is paving the way to AFM developments in medical and clinical fields, in which statistical significance of results is a prerequisite.
在过去的 20 年中,原子力显微镜(AFM)已成为生物学研究中无处不在的技术,允许在接近生理条件下分析从单个分子到活细胞的生物样品。尽管它的应用越来越广泛,但低处理吞吐量仍然是一个主要的缺点。在这里,我们提出了一种在允许全自动多样本分析的设备上验证的解决方案。我们的方法主要设计用于研究流体和生物细胞中的样品。作为概念验证,我们证明了其在完成数据处理之前应用于检测和扫描固定和活细菌的可行性。然后,对固定假结核耶尔森菌的物理参数证明了两种不同处理(即庆大霉素和加热)的效果。提出的多样本分析允许增加扫描的样本数量,同时限制用户的输入。重要的是,作为自动化过程的一部分,定期进行悬臂清洁和控制步骤,以确保一致的扫描质量。我们讨论了这种方法如何为医学和临床领域的 AFM 发展铺平道路,在这些领域中,结果的统计学意义是先决条件。