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利用胶体探针原子力显微镜对活细胞进行纳米力学和形貌成像。

Nanomechanical and topographical imaging of living cells by atomic force microscopy with colloidal probes.

作者信息

Puricelli Luca, Galluzzi Massimiliano, Schulte Carsten, Podestà Alessandro, Milani Paolo

机构信息

CIMaINa and Department of Physics, Università degli Studi di Milano, Via Celoria 16, 20133 Milano, Italy.

出版信息

Rev Sci Instrum. 2015 Mar;86(3):033705. doi: 10.1063/1.4915896.

Abstract

Atomic Force Microscopy (AFM) has a great potential as a tool to characterize mechanical and morphological properties of living cells; these properties have been shown to correlate with cells' fate and patho-physiological state in view of the development of novel early-diagnostic strategies. Although several reports have described experimental and technical approaches for the characterization of cellular elasticity by means of AFM, a robust and commonly accepted methodology is still lacking. Here, we show that micrometric spherical probes (also known as colloidal probes) are well suited for performing a combined topographic and mechanical analysis of living cells, with spatial resolution suitable for a complete and accurate mapping of cell morphological and elastic properties, and superior reliability and accuracy in the mechanical measurements with respect to conventional and widely used sharp AFM tips. We address a number of issues concerning the nanomechanical analysis, including the applicability of contact mechanical models and the impact of a constrained contact geometry on the measured Young's modulus (the finite-thickness effect). We have tested our protocol by imaging living PC12 and MDA-MB-231 cells, in order to demonstrate the importance of the correction of the finite-thickness effect and the change in Young's modulus induced by the action of a cytoskeleton-targeting drug.

摘要

原子力显微镜(AFM)作为一种表征活细胞力学和形态学特性的工具具有巨大潜力;鉴于新型早期诊断策略的发展,这些特性已被证明与细胞命运和病理生理状态相关。尽管有几篇报道描述了通过AFM表征细胞弹性的实验和技术方法,但仍然缺乏一种稳健且被普遍接受的方法。在这里,我们表明微米级球形探针(也称为胶体探针)非常适合对活细胞进行形貌和力学联合分析,其空间分辨率适合对细胞形态和弹性特性进行完整而准确的映射,并且在力学测量方面相对于传统且广泛使用的尖锐AFM探针具有更高的可靠性和准确性。我们解决了一些与纳米力学分析相关的问题,包括接触力学模型的适用性以及受限接触几何形状对测量的杨氏模量(有限厚度效应)的影响。我们通过对活的PC12和MDA - MB - 231细胞进行成像来测试我们的方案,以证明校正有限厚度效应的重要性以及细胞骨架靶向药物作用引起的杨氏模量变化。

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