Feng Yaqi, Liu Lianqing, Li Mi
State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang 110016, China.
University of Chinese Academy of Sciences, Beijing 100049, China.
ACS Appl Mater Interfaces. 2025 Aug 6;17(31):45172-45187. doi: 10.1021/acsami.5c09704. Epub 2025 Jul 22.
Mechanical forces are crucial for cellular function and disease, and particularly, atomic force microscopy (AFM)-based force spectroscopy has become a standard and important platform for characterizing the mechanical properties of single cells. Here, we present a study of micropipette-assisted AFM that enables multistate nanomechanical phenotyping of a living cell during its biological processes. Micropipette-assisted AFM offers the additional capability to manipulate single living cells in three dimensions, allowing the utilization of an AFM-based force spectroscopy assay to construct the dynamic nanomechanical phenotypes of single cells at multiple states. With micropipette manipulations, individual living cells could be selectively isolated in situ and subsequently positioned at specific locations on the engineered substrates with controllable properties. Subsequently, the mechanical changes of the same cells in the changed physiological states due to the interactions between cells and their altered microenvironments could be measured by AFM. The effectiveness of the proposed method was verified in a variety of systems, including single-cell responses to ECM biochemical cues, single-cell responses to ECM physical cues, and single-cell mechanics involved in cell-cell interactions within physical confinement, revealing numerous distinctive behaviors and nanomechanical phenotypes of individual cells. The study demonstrates an experimental approach to build the mechanical atlas of single cells undergoing regulated physiological and pathological changes, which offers additional possibilities for dissecting cellular heterogeneity from the biomechanical perspective and will benefit mechanobiology.
机械力对细胞功能和疾病至关重要，特别是基于原子力显微镜（AFM）的力谱技术已成为表征单细胞力学特性的标准且重要的平台。在此，我们展示了一项关于微吸管辅助AFM的研究，该技术能够在活细胞的生物过程中对其进行多状态纳米力学表型分析。微吸管辅助AFM提供了在三维空间中操纵单个活细胞的额外能力，使得基于AFM的力谱分析能够构建单个细胞在多个状态下的动态纳米力学表型。通过微吸管操作，可以在原位选择性地分离单个活细胞，随后将其定位在具有可控特性的工程化基质上的特定位置。随后，可以通过AFM测量同一细胞在由于细胞与其改变的微环境之间的相互作用而导致的生理状态变化时的力学变化。该方法的有效性在多种系统中得到了验证，包括单细胞对细胞外基质生化信号的反应、单细胞对细胞外基质物理信号的反应以及物理限制内细胞间相互作用中涉及的单细胞力学，揭示了单个细胞的许多独特行为和纳米力学表型。这项研究展示了一种构建经历生理和病理变化调节的单细胞力学图谱的实验方法，这为从生物力学角度剖析细胞异质性提供了更多可能性，并将有益于力学生物学。
