The Emergence of AFM Applications to Cell Biology: How new technologies are facilitating investigation of human cells in health and disease at the nanoscale.
The Emergence of AFM Applications to Cell Biology: How new technologies are facilitating investigation of human cells in health and disease at the nanoscale.
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发表时间：
2011
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通讯作者：
Ruiguo Yang;N. Xi;C. Fung;K. Seiffert-Sinha;K. Lai;A. Sinha
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作者：
Ruiguo Yang;N. Xi;C. Fung;K. Seiffert-Sinha;K. Lai;A. Sinha
基于原子力显微镜（AFM）的纳米机器人技术已用于半导体纳米器件制造近十年。利用该技术无与伦比的精确定位能力，如今在生物环境中越来越多地进行高分辨率成像和力学性能表征。AFM还为在纳米尺度上处理和操纵生物材料提供了前景。它相对于其他方法具有独特优势，允许在能维持生理条件的液相中进行实验。利用这些特性，我们的团队通过将AFM针尖与细胞成分的相互作用力控制在纳牛或亚纳牛范围内，观察到了活细胞的细胞膜和细胞骨架结构。通过对人角质形成细胞在特异性抗体处理前后的杨氏模量值进行统计分析，观察到了细胞刚度的变化。此外，我们将AFM悬臂用作机械臂进行推、拉和切割操作，对细胞相关结构进行纳米尺度的操纵。在细胞上进行了AFM引导的纳米切割，即纳米手术，通过分辨率低于100纳米的切割来切断连接相邻角质形成细胞的中间丝。最后，我们使用功能化的AFM针尖探测细胞表面受体以获得结合力测量值。该技术构成了单分子力谱（SMFS）的基础。除了增进我们对细胞生物学中动态信号事件的基本理解外，基于AFM的生物医学研究的这些进展有望促进与疾病诊断进展和治疗相关的生物标志物的寻找。
Atomic Force Microscopy (AFM) based nanorobotics has been used for building nano devices in semiconductors for almost a decade. Leveraging the unparallel precision localization capabilities of this technology, high resolution imaging and mechanical property characterization is now increasingly being performed in biological settings. AFM also offers the prospect for handling and manipulating biological materials at nanometer scale. It has unique advantages over other methods, permitting experiments in the liquid phase where physiological conditions can be maintained. Taking advantage of these properties, our group has visualized membrane and cytoskeletal structures of live cells by controlling the interaction force of the AFM tip with cellular components at the nN or sub-nN range. Cell stiffness changes were observed by statistically analyzing the Young's modulus values of human keratinocytes before and after specific antibody treatment. Furthermore, we used the AFM cantilever as a robotic arm for mechanical pushing, pulling and cutting to perform nanoscale manipulations of cell-associated structures. AFM guided nano-dissection, or nanosurgery was enacted on the cell in order to sever intermediate filaments connecting neighboring keratinocytes via sub 100 nm resolution cuts. Finally, we have used a functionalized AFM tip to probe cell surface receptors to obtain binding force measurements. This technique formed the basis for Single Molecule Force Spectroscopy (SMFS). In addition to enhancing our basic understanding of dynamic signaling events in cell biology, these advancements in AFM based biomedical investigations can be expected to facilitate the search for biomarkers related to disease diagnosis progress and treatment.