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Probing cytoskeletal structures by coupling optical superresolution and AFM techniques for a correlative approach

机译:通过结合光学超分辨率和AFM技术来探索细胞骨架结构的相关方法

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摘要

In this article, we describe and show the application of some of the most advanced fluorescence superresolution techniques, STED AFM and STORM AFM microscopy towards imaging of cytoskeletal structures, such as microtubule filaments. Mechanical and structural properties can play a relevant role in the investigation of cytoskeletal structures of interest, such as microtubules, that provide support to the cell structure. In fact, the mechanical properties, such as the local stiffness and the elasticity, can be investigated by AFM force spectroscopy with tens of nanometers resolution. Force curves can be analyzed in order to obtain the local elasticity (and the Young's modulus calculation by fitting the force curves from every pixel of interest), and the combination with STED/STORM microscopy integrates the measurement with high specificity and yields superresolution structural information. This hybrid modality of superresolution-AFM working is a clear example of correlative multimodal microscopy.
机译:在本文中,我们描述并显示了一些最先进的荧光超分辨率技术,STED AFM和STORM AFM显微镜在细胞骨架结构(如微管细丝)成像中的应用。机械和结构性质可以在研究感兴趣的细胞骨架结构(例如微管)中发挥重要作用,这些结构为细胞结构提供了支持。实际上,可以通过数十纳米分辨率的AFM力谱研究机械性能,例如局部刚度和弹性。可以分析力曲线以获得局部弹性(并通过拟合来自每个感兴趣像素的力曲线来计算杨氏模量),并且与STED / STORM显微镜的结合将测量具有高度的特异性集成在一起,并产生超分辨率的结构信息。超分辨率-AFM工作的这种混合形式是相关多峰显微镜的一个明显例子。

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