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Journal Article

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

MPS-Authors

Gavrilov,  Alina
Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society;

Mihlan,  Michael
Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society;

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Lämmermann,  Tim
Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society;

External Resource

https://www.nature.com/articles/s41467-022-29091-0
(Publisher version)

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Fulltext (public)

Jünger et al. 2022.pdf
(Publisher version), 7MB

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Citation

Jünger, F., Ruh, D., Strobel, D., Michiels, R., Huber, D., Brandel, A., et al. (2022). 100 Hz ROCS microscopy correlated with fluorescence reveals cellular dynamics on different spatiotemporal scales. Nature Communications, 13: 1758. doi:10.1038/s41467-022-29091-0.


Cite as: https://hdl.handle.net/21.11116/0000-000A-31C9-C
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.