Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy

Vytautas Navikas, Samuel M. Leitao, Kristin S. Grussmayer, Adrien Descloux, Barney Drake, Klaus Yserentant, Philipp Werther, Dirk-Peter Herten, Richard Wombacher, Aleksandra Radenovic () and Georg E. Fantner ()
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Vytautas Navikas: Swiss Federal InstSIitute of Technology Lausanne (EPFL)
Samuel M. Leitao: Swiss Federal Institute of Technology Lausanne (EPFL)
Kristin S. Grussmayer: Swiss Federal InstSIitute of Technology Lausanne (EPFL)
Adrien Descloux: Swiss Federal InstSIitute of Technology Lausanne (EPFL)
Barney Drake: Swiss Federal Institute of Technology Lausanne (EPFL)
Klaus Yserentant: University of Birmingham
Philipp Werther: Heidelberg University
Dirk-Peter Herten: University of Birmingham
Richard Wombacher: Heidelberg University
Aleksandra Radenovic: Swiss Federal InstSIitute of Technology Lausanne (EPFL)
Georg E. Fantner: Swiss Federal Institute of Technology Lausanne (EPFL)

Nature Communications, 2021, vol. 12, issue 1, 1-9

Abstract: Abstract High-resolution live-cell imaging is necessary to study complex biological phenomena. Modern fluorescence microscopy methods are increasingly combined with complementary, label-free techniques to put the fluorescence information into the cellular context. The most common high-resolution imaging approaches used in combination with fluorescence imaging are electron microscopy and atomic-force microscopy (AFM), originally developed for solid-state material characterization. AFM routinely resolves atomic steps, however on soft biological samples, the forces between the tip and the sample deform the fragile membrane, thereby distorting the otherwise high axial resolution of the technique. Here we present scanning ion-conductance microscopy (SICM) as an alternative approach for topographical imaging of soft biological samples, preserving high axial resolution on cells. SICM is complemented with live-cell compatible super-resolution optical fluctuation imaging (SOFI). To demonstrate the capabilities of our method we show correlative 3D cellular maps with SOFI implementation in both 2D and 3D with self-blinking dyes for two-color high-order SOFI imaging. Finally, we employ correlative SICM/SOFI microscopy for visualizing actin dynamics in live COS-7 cells with subdiffraction-resolution.

Date: 2021
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DOI: 10.1038/s41467-021-24901-3

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