Plodinec Marija, Lim Roderick Y H
Biozentrum and The Swiss Nanoscience Institute, University of Basel, Room 304B, Klingelbergstrasse 50/70, 4056, Basel, Switzerland,
Methods Mol Biol. 2015;1293:231-46. doi: 10.1007/978-1-4939-2519-3_14.
The mechanical properties of living cells and tissues are important for a variety of functional processes in vivo, including cell adhesion, migration, proliferation and differentiation. Changes in mechano-cellular phenotype, for instance, are associated with cancer progression. Atomic force microscopy (AFM) is an enabling technique that topographically maps and quantifies the mechanical properties of complex biological matter in physiological aqueous environments at the nanometer length scale. Recently we applied AFM to spatially resolve the distribution of nanomechanical stiffness across human breast cancer biopsies in comparison to healthy tissue and benign tumors. This led to the finding that AFM provides quantitative mechano-markers that may have translational significance for the clinical diagnosis of cancer. Here, we provide a comprehensive description of sample preparation methodology, instrumentation, data acquisition and analysis that allows for the quantitative nanomechanical profiling of unadulterated tissue at submicron spatial resolution and nano-Newton (nN) force sensitivity in physiological conditions.
活细胞和组织的力学特性对于体内多种功能过程至关重要，包括细胞黏附、迁移、增殖和分化。例如，机械细胞表型的变化与癌症进展相关。原子力显微镜（AFM）是一种能够在生理水环境中以纳米长度尺度对复杂生物物质的机械特性进行形貌映射和量化的技术。最近，我们应用AFM在空间上解析了人类乳腺癌活检组织与健康组织和良性肿瘤相比的纳米力学刚度分布。这一研究发现AFM能够提供定量的力学标志物，可能对癌症的临床诊断具有转化意义。在此，我们全面描述了样本制备方法、仪器设备、数据采集和分析，这些方法能够在生理条件下以亚微米空间分辨率和纳牛顿（nN）力灵敏度对未掺杂组织进行定量纳米力学分析。
