Confocal interferometric scattering microscopy reveals 3D nanoscopic structure and dynamics in live cells

Michelle Küppers, David Albrecht, Anna D. Kashkanova, Jennifer Lühr and Vahid Sandoghdar ()
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Michelle Küppers: Max Planck Institute for the Science of Light
David Albrecht: Max Planck Institute for the Science of Light
Anna D. Kashkanova: Max Planck Institute for the Science of Light
Jennifer Lühr: Max Planck Institute for the Science of Light
Vahid Sandoghdar: Max Planck Institute for the Science of Light

Nature Communications, 2023, vol. 14, issue 1, 1-12

Abstract: Abstract Bright-field light microscopy and related phase-sensitive techniques play an important role in life sciences because they provide facile and label-free insights into biological specimens. However, lack of three-dimensional imaging and low sensitivity to nanoscopic features hamper their application in many high-end quantitative studies. Here, we demonstrate that interferometric scattering (iSCAT) microscopy operated in the confocal mode provides unique label-free solutions for live-cell studies. We reveal the nanometric topography of the nuclear envelope, quantify the dynamics of the endoplasmic reticulum, detect single microtubules, and map nanoscopic diffusion of clathrin-coated pits undergoing endocytosis. Furthermore, we introduce the combination of confocal and wide-field iSCAT modalities for simultaneous imaging of cellular structures and high-speed tracking of nanoscopic entities such as single SARS-CoV-2 virions. We benchmark our findings against simultaneously acquired fluorescence images. Confocal iSCAT can be readily implemented as an additional contrast mechanism in existing laser scanning microscopes. The method is ideally suited for live studies on primary cells that face labeling challenges and for very long measurements beyond photobleaching times.

Date: 2023
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DOI: 10.1038/s41467-023-37497-7

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