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原位原子力显微镜检测原位暴露细胞核的硬度。

In situ AFM detection of the stiffness of the in situ exposed cell nucleus.

机构信息

Jiangxi Key Laboratory for Microscale Interdisciplinary Study, Institute for Advanced Study, Nanchang University, Nanchang, Jiangxi 330031, PR China.

Jiangxi Key Laboratory for Microscale Interdisciplinary Study, Institute for Advanced Study, Nanchang University, Nanchang, Jiangxi 330031, PR China; College of Life Sciences, Nanchang University, Nanchang, Jiangxi 330031, PR China.

出版信息

Biochim Biophys Acta Mol Cell Res. 2021 Apr;1868(5):118985. doi: 10.1016/j.bbamcr.2021.118985. Epub 2021 Feb 15.

Abstract

Biomechanical properties of the cell nucleus play critical roles in cell behaviors and functions. As one important biomechanical property, the stiffness (or Young's modulus) of the cell nucleus has been widely investigated by different techniques including atomic force microscopy (AFM). In most of previous studies, the stiffness of the nuclear region of an intact cell or the stiffness of the isolated nucleus was detected. In this study, we developed a strategy for in situ detecting the stiffness of the cell nucleus via AFM. The extranuclear components of adherent cells (endothelial cells) were in situ removed by Triton X-100 treatment and the bare, adherent nuclei were exposed for in situ AFM force measurement. We found that the nuclear regions of intact cells (5.59 ± 1.55 kPa) had a relatively higher average Young's modulus than the nonnuclear regions (1.47 ± 0.77 kPa) and that the in situ exposed nuclei (22.06 ± 7.29 kPa) were much stiffer than the nuclear regions of intact cells. This strategy is very simple and effective for detecting the stiffness of the cell nucleus and potentially is promising for a wide application.

摘要

细胞核的生物力学特性在细胞行为和功能中起着关键作用。作为一个重要的生物力学特性,细胞核的硬度(或杨氏模量)已被多种技术广泛研究,包括原子力显微镜(AFM)。在大多数先前的研究中,检测了完整细胞的核区的硬度或分离核的硬度。在这项研究中,我们开发了一种通过 AFM 原位检测细胞核硬度的策略。通过 Triton X-100 处理原位去除贴壁细胞(内皮细胞)的核外成分,并暴露裸贴壁核进行原位 AFM 力测量。我们发现完整细胞的核区(5.59±1.55kPa)的平均杨氏模量相对高于非核区(1.47±0.77kPa),并且原位暴露的核(22.06±7.29kPa)比完整细胞的核区硬得多。这种策略非常简单有效,可用于检测细胞核的硬度,具有广泛的应用前景。

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