High-speed atomic force microscopy imaging of live mammalian cells

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Author(s)

    • Shibata Mikihiro
    • High-speed AFM for Biological Application Unit, Institute for Frontier Science Initiative, Kanazawa University|Bio-AFM Frontier Research Center, Kanazawa University
    • 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

Abstract

<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>

Journal

  • Biophysics and Physicobiology

    Biophysics and Physicobiology 14(0), 127-135, 2017

    The Biophysical Society of Japan

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