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

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  • Michelle Küppers

    (Max Planck Institute for the Science of Light
    Max-Planck-Zentrum für Physik und Medizin
    Friedrich-Alexander-Universität Erlangen-Nürnberg)

  • David Albrecht

    (Max Planck Institute for the Science of Light
    Max-Planck-Zentrum für Physik und Medizin)

  • Anna D. Kashkanova

    (Max Planck Institute for the Science of Light
    Max-Planck-Zentrum für Physik und Medizin)

  • Jennifer Lühr

    (Max Planck Institute for the Science of Light
    Max-Planck-Zentrum für Physik und Medizin)

  • Vahid Sandoghdar

    (Max Planck Institute for the Science of Light
    Max-Planck-Zentrum für Physik und Medizin
    Friedrich-Alexander-Universität Erlangen-Nürnberg)

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.

Suggested Citation

  • Michelle Küppers & David Albrecht & Anna D. Kashkanova & Jennifer Lühr & Vahid Sandoghdar, 2023. "Confocal interferometric scattering microscopy reveals 3D nanoscopic structure and dynamics in live cells," Nature Communications, Nature, vol. 14(1), pages 1-12, December.
    • Handle: RePEc:nat:natcom:v:14:y:2023:i:1:d:10.1038_s41467-023-37497-7
    DOI: 10.1038/s41467-023-37497-7
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    References listed on IDEAS

    as
    1. Marek Piliarik & Vahid Sandoghdar, 2014. "Direct optical sensing of single unlabelled proteins and super-resolution imaging of their binding sites," Nature Communications, Nature, vol. 5(1), pages 1-8, December.
    2. Tan H. Nguyen & Mikhail E. Kandel & Marcello Rubessa & Matthew B. Wheeler & Gabriel Popescu, 2017. "Gradient light interference microscopy for 3D imaging of unlabeled specimens," Nature Communications, Nature, vol. 8(1), pages 1-9, December.
    3. Charlotte Pain & Verena Kriechbaumer & Maike Kittelmann & Chris Hawes & Mark Fricker, 2019. "Quantitative analysis of plant ER architecture and dynamics," Nature Communications, Nature, vol. 10(1), pages 1-15, December.
    4. Vijay Raj Singh & Yi An Yang & Hanry Yu & Roger D. Kamm & Zahid Yaqoob & Peter T. C. So, 2019. "Studying nucleic envelope and plasma membrane mechanics of eukaryotic cells using confocal reflectance interferometric microscopy," Nature Communications, Nature, vol. 10(1), pages 1-8, December.
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