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100 Hz ROCS microscopy correlated with fluorescence reveals cellular dynamics on different spatiotemporal scales

Author

Listed:
  • Felix Jünger

    (University of Freiburg)

  • Dominic Ruh

    (University of Freiburg)

  • Dominik Strobel

    (University of Freiburg)

  • Rebecca Michiels

    (University of Freiburg)

  • Dominik Huber

    (University of Freiburg)

  • Annette Brandel

    (University of Freiburg
    University of Freiburg)

  • Josef Madl

    (University of Freiburg
    University of Freiburg
    German Collaborative Research Centre SFB1425, University of Freiburg)

  • Alina Gavrilov

    (Max Planck Institute of Immunobiology and Epigenetics)

  • Michael Mihlan

    (Max Planck Institute of Immunobiology and Epigenetics)

  • Caterina Cora Daller

    (University of Freiburg)

  • Eva A. Rog-Zielinska

    (University of Freiburg
    German Collaborative Research Centre SFB1425, University of Freiburg)

  • Winfried Römer

    (University of Freiburg
    University of Freiburg
    University of Freiburg)

  • Tim Lämmermann

    (Max Planck Institute of Immunobiology and Epigenetics)

  • Alexander Rohrbach

    (University of Freiburg
    University of Freiburg
    University of Freiburg
    German Collaborative Research Centre SFB1425, University of Freiburg)

Abstract

Fluorescence techniques dominate the field of live-cell microscopy, but bleaching and motion blur from too long integration times limit dynamic investigations of small objects. High contrast, label-free life-cell imaging of thousands of acquisitions at 160 nm resolution and 100 Hz is possible by Rotating Coherent Scattering (ROCS) microscopy, where intensity speckle patterns from all azimuthal illumination directions are added up within 10 ms. In combination with fluorescence, we demonstrate the performance of improved Total Internal Reflection (TIR)-ROCS with variable illumination including timescale decomposition and activity mapping at five different examples: millisecond reorganization of macrophage actin cortex structures, fast degranulation and pore opening in mast cells, nanotube dynamics between cardiomyocytes and fibroblasts, thermal noise driven binding behavior of virus-sized particles at cells, and, bacterial lectin dynamics at the cortex of lung cells. Using analysis methods we present here, we decipher how motion blur hides cellular structures and how slow structure motions cover decisive fast motions.

Suggested Citation

  • Felix Jünger & Dominic Ruh & Dominik Strobel & Rebecca Michiels & Dominik Huber & Annette Brandel & Josef Madl & Alina Gavrilov & Michael Mihlan & Caterina Cora Daller & Eva A. Rog-Zielinska & Winfrie, 2022. "100 Hz ROCS microscopy correlated with fluorescence reveals cellular dynamics on different spatiotemporal scales," Nature Communications, Nature, vol. 13(1), pages 1-17, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-29091-0
    DOI: 10.1038/s41467-022-29091-0
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    References listed on IDEAS

    as
    1. Mikhail E. Kandel & Chenfei Hu & Ghazal Naseri Kouzehgarani & Eunjung Min & Kathryn Michele Sullivan & Hyunjoon Kong & Jennifer M. Li & Drew N. Robson & Martha U. Gillette & Catherine Best-Popescu & G, 2019. "Epi-illumination gradient light interference microscopy for imaging opaque structures," Nature Communications, Nature, vol. 10(1), pages 1-9, December.
    2. Stylianos Bournazos & Davide Corti & Herbert W. Virgin & Jeffrey V. Ravetch, 2020. "Fc-optimized antibodies elicit CD8 immunity to viral respiratory infection," Nature, Nature, vol. 588(7838), pages 485-490, December.
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