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无标记、非侵入式单细胞水平微生物表面超微结构分析。

Label-free and noninvasive analysis of microorganism surface epistructures at the single-cell level.

机构信息

Department of Engineering and System Science, National Tsing Hua University, Hsinchu, Taiwan; Université Paris Saclay, ENS Paris Saclay, CNRS Institut d'Alembert, SATIE, Gif sur Yvette, France.

Department of Mechanical Engineering, National Taiwan University, Taipei, Taiwan.

出版信息

Biophys J. 2023 May 16;122(10):1794-1806. doi: 10.1016/j.bpj.2023.04.012. Epub 2023 Apr 11.

Abstract

Cell surface properties of microorganisms provide abundant information for their physiological status and fate choice. However, current methods for analyzing cell surface properties require labeling or fixation, which can alter the cell activity. This study establishes a label-free, rapid, noninvasive, and quantitative analysis of cell surface properties, including the presence and the dimension of epistructure, down to the single-cell level and at the nanometer scale. Simultaneously, electrorotation provides dielectric properties of intracellular contents. With the combined information, the growth phase of microalgae cells can be identified. The measurement is based on electrorotation of single cells, and an electrorotation model accounting for the surface properties is developed to properly interpret experimental data. The epistructure length measured by electrorotation is validated by scanning electron microscopy. The measurement accuracy is satisfactory in particular in the case of microscale epistructures in the exponential phase and nanoscale epistructures in the stationary phase. However, the measurement accuracy for nanoscale epistructures on cells in the exponential phase is offset by the effect of a thick double layer. Lastly, a diversity in epistructure length distinguishes exponential phase from stationary phase.

摘要

微生物的表面特性为其生理状态和命运选择提供了丰富的信息。然而,目前分析细胞表面特性的方法需要进行标记或固定,这可能会改变细胞的活性。本研究建立了一种无需标记、快速、非侵入性且定量的单细胞和纳米尺度的细胞表面特性分析方法,包括微结构的存在和尺寸。同时,电动旋转提供了细胞内物质的介电特性。结合这些信息,可以识别微藻细胞的生长阶段。该测量基于单细胞的电动旋转,开发了一个考虑表面特性的电动旋转模型,以正确解释实验数据。通过扫描电子显微镜验证了电动旋转测量的微结构长度。在指数期的微结构和静止期的纳米结构的情况下,测量精度令人满意。然而,在指数期的纳米结构的情况下,由于双层的影响,测量精度会受到影响。最后,微结构长度的多样性可以区分指数期和静止期。

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