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; Georg E. Fantner

doi:10.1038/s41467-021-24901-3

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, Georg E. Fantner

Cardiovascular Sciences

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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.

Original language	English
Article number	4565
Pages (from-to)	4565
Journal	Nature Communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-24901-3
Publication status	Published - 27 Jul 2021

Bibliographical note

Funding Information:
V.N. and A.R. acknowledges the support of the Max Planck-EPFL Center for Molecular Nanoscience and Technology and Swiss National Science Foundation through the National Centre of Competence in Research Bio-Inspired Materials. K.Y. and D.P.H. gratefully acknowledge funding by the Centre of Membrane Proteins and Receptors (COMPARE, Universities of Birmingham and Nottingham). G.E.F. acknowledges the support by the European Research Council under grant number ERC-2017-CoG; InCell; Project number 773091. The authors thank Adrian Pascal Nievergelt and Charlène Brillard for their input on technical development of SICM controller. The authors also thank Prof. Jens Anders and his team from the University of Stuttgart for insightful discussions into high-speed transimpedance amplifiers.

Publisher Copyright:
© 2021, The Author(s).

ASJC Scopus subject areas

Chemistry(all)
Biochemistry, Genetics and Molecular Biology(all)
Physics and Astronomy(all)

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navikasv2021correlativeFinal published version, 11.6 MBLicence: Creative Commons: Attribution (CC BY)

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Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy
Navikas, V., Leitao, S. M., Grussmayer, K. S., Descloux, A., Drake, B., Yserentant, K., Werther, P., Herten, D-P., Wombacher, R., Radenovic, A. & Fantner, G. E., 9 Nov 2020.
Research output: Working paper/Preprint › Preprint

Cite this

@article{0c6456015c4f45809767c26897b91077,

title = "Correlative 3D microscopy of single cells using super-resolution and scanning ion-conductance microscopy",

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.",

author = "Vytautas Navikas and Leitao, {Samuel M.} and Grussmayer, {Kristin S.} and Adrien Descloux and Barney Drake and Klaus Yserentant and Philipp Werther and Dirk-Peter Herten and Richard Wombacher and Aleksandra Radenovic and Fantner, {Georg E.}",

note = "Funding Information: V.N. and A.R. acknowledges the support of the Max Planck-EPFL Center for Molecular Nanoscience and Technology and Swiss National Science Foundation through the National Centre of Competence in Research Bio-Inspired Materials. K.Y. and D.P.H. gratefully acknowledge funding by the Centre of Membrane Proteins and Receptors (COMPARE, Universities of Birmingham and Nottingham). G.E.F. acknowledges the support by the European Research Council under grant number ERC-2017-CoG; InCell; Project number 773091. The authors thank Adrian Pascal Nievergelt and Charl{\`e}ne Brillard for their input on technical development of SICM controller. The authors also thank Prof. Jens Anders and his team from the University of Stuttgart for insightful discussions into high-speed transimpedance amplifiers. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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AU - Grussmayer, Kristin S.

AU - Descloux, Adrien

AU - Drake, Barney

AU - Yserentant, Klaus

AU - Werther, Philipp

AU - Herten, Dirk-Peter

AU - Wombacher, Richard

AU - Radenovic, Aleksandra

AU - Fantner, Georg E.

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N2 - 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.

AB - 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.

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JF - Nature Communications

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ER -

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

Abstract

Bibliographical note

ASJC Scopus subject areas

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Research output

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

Cite this