Long-tip high-speed atomic force microscopy for nanometer-scale imaging in live cells

Scientific Reports
Mikihiro ShibataRyohei Yasuda

Abstract

Visualization of morphological dynamics of live cells with nanometer resolution under physiological conditions is highly desired, but challenging. It has been demonstrated that high-speed atomic force microscopy is a powerful technique for visualizing dynamics of biomolecules under physiological conditions. However, application of high-speed atomic force microscopy for imaging larger objects such as live mammalian cells has been complicated because of the collision between the cantilever and samples. Here, we demonstrate that attaching an extremely long (~3 μm) and thin (~5 nm) tip by amorphous carbon to the cantilever allows us to image the surface structure of live cells with the spatiotemporal resolution of nanometers and seconds. We demonstrate that long-tip high-speed atomic force microscopy is capable of imaging morphogenesis of filopodia, membrane ruffles, pit formation, and endocytosis in COS-7, HeLa cells and hippocampal neurons.

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Citations

Nov 2, 2015·Ultramicroscopy·Takayuki UchihashiToshio Ando
Dec 18, 2015·Scientific Reports·Yu YanLijie Qiao
Jul 15, 2015·Scientific Reports·Adrian P NievergeltGeorg E Fantner
Jul 3, 2015·The Review of Scientific Instruments·Shingo FukudaToshio Ando
May 26, 2016·Biophysical Journal·Andrew ShevchukYuri E Korchev
Aug 24, 2016·Proceedings of the National Academy of Sciences of the United States of America·Padmini RangamaniGeorge Oster
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Jan 28, 2016·Scientific Reports·Naikun GaoDuo Liu
Apr 7, 2017·Nature Nanotechnology·Yves F DufrêneDaniel J Müller
Dec 20, 2017·Biophysics Reviews·Toshio Ando
Oct 3, 2018·International Journal of Molecular Sciences·Adrian P NievergeltGeorg E Fantner
Apr 16, 2020·Nanoscale·Pablo D GarciaRicardo Garcia
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Feb 26, 2019·Journal of Physics D: Applied Physics·Toshio AndoGiovanni Zifarelli
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Jul 1, 2019·The Review of Scientific Instruments·Lu LiuXiaotang Hu
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Jul 8, 2017·Scientific Reports·Naikun GaoDuo Liu
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Nov 23, 2020·Journal of Molecular Biology·Daniël P Melters, Yamini Dalal
Dec 17, 2019·Analytical Chemistry·Yasufumi TakahashiTakeshi Fukuma

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Methods Mentioned

BETA
AFM
force measurements
optical microscopy
fluorescence microscopy
fluorescence imaging
fluorescence resonance
transfection

Software Mentioned

AFM
HS

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