High-speed atomic force microscopy imaging of live mammalian cells

Shibata, Mikihiro; Watanabe, Hiroki; Uchihashi, Takayuki; Ando, Toshio; Yasuda, Ryohei

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High-speed atomic force microscopy imaging of live mammalian cells

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Shibata, Mikihiro
Max Planck Florida Institute for Neuroscience, Max Planck Society;

Yasuda, Ryohei
Max Planck Florida Institute for Neuroscience, Max Planck Society;

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Citation

Shibata, M., Watanabe, H., Uchihashi, T., Ando, T., & Yasuda, R. (2017). High-speed atomic force microscopy imaging of live mammalian cells. Biophysics and Physicobiology, 127-135. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5590786/.

Cite as: https://hdl.handle.net/21.11116/0000-0003-D520-8

Abstract

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.