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Brillouin light scattering anisotropy microscopy for imaging the viscoelastic anisotropy in living cells

Nature Photonics volume 18, pages 276–285 (2024)Cite this article

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Abstract

Maintaining and modulating mechanical anisotropy is essential for biological processes. However, how this is achieved at the microscopic scale in living soft matter is not always clear. Although Brillouin light scattering (BLS) spectroscopy can probe the mechanical properties of materials, spatiotemporal mapping of mechanical anisotropies in living matter with BLS microscopy has been complicated by the need for sequential measurements with tilted excitation and detection angles. Here we introduce Brillouin light scattering anisotropy microscopy (BLAM) for mapping high-frequency viscoelastic anisotropy inside living cells. BLAM employs a radial virtually imaged phased array that enables the collection of angle-resolved dispersion in a single shot, thus enabling us to probe phonon modes in living matter along different directions simultaneously. We demonstrate a precision of 10 MHz in the determination of the Brillouin frequency shift, at a spatial resolution of 2 µm. Following proof-of-principle experiments on muscle myofibres, we apply BLAM to the study of two fundamental biological processes. In plant cell walls, we observe a switch from anisotropic to isotropic wall properties that may lead to asymmetric growth. In mammalian cell nuclei, we uncover a spatiotemporally oscillating elastic anisotropy correlated to chromatin condensation. Our results highlight the role that high-frequency mechanics can play in the regulation of diverse fundamental processes in biological systems. We expect BLAM to find diverse applications in biomedical imaging and material characterization.

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Fig. 1: Brillouin light scattering anisotropy microscopy (BLAM).
Fig. 2: BLS anisotropy of A. thaliana hypocotyl cell walls is perturbed in pme5o mutants.
Fig. 3: Cell nuclei have a spatially heterogeneous and locally oscillating BLS anisotropy.
Fig. 4: Scaling of the BLS anisotropy in cell nuclei reveals long-range correlations.

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Data availability

Data for this study are available in the main text or Supplementary Information, and where not the case, available at https://doi.org/10.5281/zenodo.10465950.

Code availability

Code used for the analysis of data that has not already been published elsewhere is available at https://doi.org/10.5281/zenodo.10465950.

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Acknowledgements

We acknowledge support from the Vienna Biocenter Core Facilities (Plant Science Facility and Advanced Microscopy Facility), and thank A. Dammermann for critical reading of the manuscript. H.K., M.U. and K.E. acknowledge funding from EU Interreg grants nos. V-A AT-CZ, RIAT-CZ and ATCZ40, the City of Vienna and the Austrian Ministry of Science (Vision 2020) and CEITEC Nano/CzechNanoLab Research Infrastructure funded by MEYS CR (LM2023051). J.M.P., W.J.W. and K.E. acknowledge funding from the Medical University of Vienna. D.C. and J.M.P. acknowledge funding from the Austrian Academy of Science. H.K. acknowledges funding from a Marie Skłodowska-Curie Action Individual Fellowship (H2020-MSCA-IF-2020, 101032071) and an Adolf-Martens fellowship from BAM. A.P. acknowledges funding from the project ANR-17-CE13-0007 ANR ‘GoodVibrations’. J.M.P. acknowledges funding from the T. von Zastrow Foundation, the Canada 150 Research Chairs Program F18-01336 and the German Federal Ministry of Education and Research (BMBF) under the project ‘Microbial Stargazing—Erforschung von Resilienzmechanismen von Mikroben und Menschen’ (ref. 01KX2324). K.E. acknowledges funding from the Austrian Science Fund—FWF (P34783).

Author information

Author notes

  1. These authors contributed equally: Hamid Keshmiri, Domagoj Cikes.

Authors and Affiliations

  1. Advanced Microscopy, Vienna Biocenter Core Facilities, Vienna Biocenter, Vienna, Austria

    Hamid Keshmiri & Kareem Elsayad

  2. Federal Institute for Materials Research and Testing, Berlin, Germany

    Hamid Keshmiri

  3. Institute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna Biocenter, Vienna, Austria

    Domagoj Cikes & Josef Penninger

  4. Institute for Physiology and Pathophysiology, Johann Kepler University, Linz, Austria

    Domagoj Cikes

  5. Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic

    Marketa Samalova

  6. Center for Molecular Biology, University of Vienna, Vienna, Austria

    Lukas Schindler & Ivan Yudushkin

  7. Max Perutz Labs, Medical University of Vienna, Vienna, Austria

    Lisa-Marie Appel & Dea Slade

  8. CEITEC BUT, Brno University of Technology, Brno, Czech Republic

    Michal Urbanek

  9. Division of Anatomy, Center for Anatomy & Cell Biology, Medical University of Vienna, Vienna, Austria

    Wolfgang J. Weninger & Kareem Elsayad

  10. Medical Imaging Cluster (MIC), Medical University of Vienna, Vienna, Austria

    Wolfgang J. Weninger & Kareem Elsayad

  11. Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), Versailles, France

    Alexis Peaucelle

  12. Helmholtz Centre for Infection Research, Braunschweig, Germany

    Josef Penninger

  13. Department of Laboratory Medicine, Medical University of Vienna, Vienna, Austria

    Josef Penninger

  14. Department of Medical Genetics, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada

    Josef Penninger

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  1. Hamid Keshmiri
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Contributions

Conceptualization and project administration were provided by K.E., methodology by H.K., D.C., M.S., L.S., L.-M.A., M.U., I.Y., D.S., W.J.W., A.P., J.P. and K.E., investigation and visualization by H.K., D.C., M.S., L.S., L.-M.A. and K.E., funding acquisition by M.U. and K.E. and supervision by M.U., I.Y., D.S., J.P. and K.E. The original draft was written by K.E. and I.Y., with review and editing carried out by all authors.

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Correspondence to Kareem Elsayad.

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Keshmiri, H., Cikes, D., Samalova, M. et al. Brillouin light scattering anisotropy microscopy for imaging the viscoelastic anisotropy in living cells. Nat. Photon. 18, 276–285 (2024). https://doi.org/10.1038/s41566-023-01368-w

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