Single-cell parallel plate mechanics by side-view optical microscopy-assisted atomic force microscopy†

IF 4.6 3区 材料科学 Q2 CHEMISTRY, MULTIDISCIPLINARY
Mingyang Yang, Yanqi Yang, Lianqing Liu and Mi Li
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Abstract

The mechanical forces of individual cells are crucial for physiological and pathological processes, and particularly atomic force microscopy (AFM)-based force spectroscopy has become an important and widely used tool for single-cell mechanical measurements. Here, we present a single-cell parallel plate mechanical measurement method based on side-view optical microscopy-assisted AFM by using a wedged probe, which enables visualization of the uniaxial AFM force spectroscopy process of the cell in real time from the vertical cross-sectional view. With this method, AFM whole-cell compression assays with uniaxial loading forces were performed to quantify the Young's modulus of the entire cell, and the effects of experimental parameters (ramp rate of the AFM probe) and drugs (actin inhibitors) on the measured cell Young's modulus were examined. Additionally, by attaching a living cell to the parallel plate of the wedged probe, AFM-based uniaxial single-cell force spectroscopy (SCFS) assays were performed to measure the adhesion forces of individual cells. The study illustrates a promising approach for single-cell mechanical analysis, which will benefit the application of AFM-based force spectroscopy in the field of mechanobiology to reveal additional insights into the regulatory role of mechanical cues in life activities.

Abstract Image

单细胞平行板力学的侧面光学显微镜辅助原子力显微镜。
单个细胞的机械力对生理和病理过程至关重要,特别是基于原子力显微镜(AFM)的力谱已成为单细胞力学测量的重要和广泛使用的工具。在此,我们提出了一种基于侧面光学显微镜辅助AFM的单细胞平行板力学测量方法,该方法使用楔形探针,可以从垂直截面视图实时可视化细胞的单轴AFM力谱过程。采用该方法,采用单轴加载力进行AFM全细胞压缩实验,量化整个细胞的杨氏模量,并检测实验参数(AFM探针的倾斜速率)和药物(肌动蛋白抑制剂)对测量细胞杨氏模量的影响。此外,通过将活细胞附着在楔形探针的平行板上,进行基于afm的单轴单细胞力谱(SCFS)测定,以测量单个细胞的粘附力。该研究阐明了单细胞力学分析的一种有前途的方法,这将有利于基于afm的力谱在力学生物学领域的应用,以揭示机械线索在生命活动中的调节作用。
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来源期刊
Nanoscale Advances
Nanoscale Advances Multiple-
CiteScore
8.00
自引率
2.10%
发文量
461
审稿时长
9 weeks
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