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

Journal article (2021)

Authors

Vytautas Navikas Swiss Federal Institute of Technology
Samuel M. Leitao Swiss Federal Institute of Technology
Georg E. Fantner Swiss Federal Institute of Technology
K.S. Grußmayer Swiss Federal Institute of Technology
Adrien Descloux Swiss Federal Institute of Technology
Barney Drake Swiss Federal Institute of Technology
Klaus Yserentant University of Birmingham
Philipp Werther University of Heidelberg
Dirk Peter Herten University of Birmingham
Richard Wombacher University of Heidelberg, Max Planck Institute for Medical Research
Aleksandra Radenovic Swiss Federal Institute of Technology
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Published Date

2021

Language

English

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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.