A hybrid high-speed atomic force–optical microscope for visualizing single membrane proteins on eukaryotic cells

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作者
Adai Colom
Ignacio Casuso
Felix Rico
Simon Scheuring
机构
[1] U1006 INSERM,
[2] Université Aix-Marseille,undefined
[3] Parc Scientifique et Technologique de Luminy,undefined
[4] 163 avenue de Luminy,undefined
[5] 13009 Marseille,undefined
[6] France,undefined
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High-speed atomic force microscopy is a powerful tool for studying structure and dynamics of proteins. So far, however, high-speed atomic force microscopy was restricted to well-controlled molecular systems of purified proteins. Here we integrate an optical microscopy path into high-speed atomic force microscopy, allowing bright field and fluorescence microscopy, without loss of high-speed atomic force microscopy performance. This hybrid high-speed atomic force microscopy/optical microscopy setup allows positioning of the high-speed atomic force microscopy tip with high spatial precision on an optically identified zone of interest on cells. We present movies at 960 ms per frame displaying aquaporin-0 array and single molecule dynamics in the plasma membrane of intact eye lens cells. This hybrid setup allows high-speed atomic force microscopy imaging on cells about 1,000 times faster than conventional atomic force microscopy/optical microscopy setups, and allows first time visualization of unlabelled membrane proteins on a eukaryotic cell under physiological conditions. This development advances high-speed atomic force microscopy from molecular to cell biology to analyse cellular processes at the membrane such as signalling, infection, transport and diffusion.
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