High-speed atomic force microscopy imaging of live mammalian cells

DOI Web Site 被引用文献3件 参考文献10件
  • Shibata Mikihiro
    High-speed AFM for Biological Application Unit, Institute for Frontier Science Initiative, Kanazawa University Bio-AFM Frontier Research Center, Kanazawa University
  • Watanabe Hiroki
    Research Institute of Biomolecule Metrology Co., Ltd.
  • Uchihashi Takayuki
    Department of Physics and Structural Biology Research Center, Graduate School of Science, Nagoya University
  • Ando Toshio
    Bio-AFM Frontier Research Center, Kanazawa University
  • Yasuda Ryohei
    Max Planck Florida Institute for Neuroscience

抄録

<p>Direct imaging of morphological dynamics of live mammalian cells with nanometer resolution under physiological conditions is highly expected, but yet challenging. High-speed atomic force microscopy (HS-AFM) is a unique technique for capturing biomolecules at work under near physiological conditions. However, application of HS-AFM for imaging of live mammalian cells was hard to be accomplished because of collision between a huge mammalian cell and a cantilever during AFM scanning. Here, we review our recent improvements of HS-AFM for imaging of activities of live mammalian cells without significant damage to the cell. The improvement of an extremely long (~3 μm) AFM tip attached to a cantilever enables us to reduce severe damage to soft mammalian cells. In addition, a combination of HS-AFM with simple fluorescence microscopy allows us to quickly locate the cell in the AFM scanning area. After these improvements, we demonstrate that developed HS-AFM for live mammalian cells is possible to image morphogenesis of filopodia, membrane ruffles, pits open-close formations, and endocytosis in COS-7, HeLa cells as well as hippocampal neurons.</p>

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