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Brillouin microscopy

Nature Reviews Methods Primers volume 4, Article number: 8 (2024) Cite this article

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Abstract

The field of Brillouin microscopy and imaging was established approximately 20 years ago, thanks to the development of non-scanning high-resolution optical spectrometers. Since then, the field has experienced rapid expansion, incorporating technologies from telecommunications, astrophotonics, multiplexed microscopy, quantum optics and machine learning. Consequently, these advancements have led to much-needed improvements in imaging speed, spectral resolution and sensitivity. The progress in Brillouin microscopy is driven by a strong demand for label-free and contact-free methods to characterize the mechanical properties of biomaterials at the cellular and subcellular scales. Understanding the local biomechanics of cells and tissues has become crucial in predicting cellular fate and tissue pathogenesis. This Primer aims to provide a comprehensive overview of the methods and applications of Brillouin microscopy. It includes key demonstrations of Brillouin microscopy and imaging that can serve as a reference for the existing research community and new adopters of this technology. The article concludes with an outlook, presenting the authors’ vision for future developments in this vibrant field. The Primer also highlights specific examples where Brillouin microscopy can have a transformative impact on biology and biomedicine.

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Fig. 1: Principles of Brillouin light scattering microscopy.
Fig. 2: Length scales of the Brillouin light scattering process.
Fig. 3: Characteristic parameters of Brillouin microscopy experiments.
Fig. 4: Brillouin imaging technology.
Fig. 5: Representative data obtained using Brillouin technology.
Fig. 6: Ballistic and multiple Brillouin scattering processes.
Fig. 7: A typical workflow depicting analysis of Brillouin microscopy data with modern machine learning approaches.
Fig. 8: Application of Brillouin microscopy to ocular disease diagnostics and treatment.

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Acknowledgements

The authors thank R. Schlüßler, S. Möllmert, K. Kim, T. Beck, J. Czarske, K. Elsayad, R. Prevedel, G. Antonacci, P. Török, F. Palombo, M. Mattarelli and T. Dehoux for many stimulating discussions. I.K. is supported by the Australian Research Council Centre of Excellence in Optical Microcombs for Breakthrough Science (CE230100006) and the Australian Research Council Centre of Excellence in Quantum Biotechnology (CE230100021). J.Z. is supported by the Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health (NIH) (K25HD097288, R21HD112663). G.S is supported by the National Science Foundation (DBI-1942003) and NIH (R21CA258008, R01EY028666, R01EY030063). A.B. acknowledges the support of the Israel Science Foundation (grant no. 2576/21). S.C. is supported by European Union — NextGenerationEU under the Italian Ministry of University and Research (MUR) National Innovation Ecosystem grant ECS00000041 — VITALITY — CUP B43C22000470005 and PRIN 2022 PNRR P2022RH4HH.

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Authors and Affiliations

  1. School of Mathematical and Physical Sciences, University of Technology Sydney, Sydney, New South Wales, Australia

    Irina Kabakova

  2. Department of Biomedical Engineering, Wayne State University, Detroit, MI, USA

    Jitao Zhang

  3. Department of Metabolism, Digestion & Reproduction, Imperial College London, London, UK

    Yuchen Xiang

  4. Istituto Officina dei Materiali–National Research Council (IOM-CNR)–Research Unit in Perugia, c/o Department of Physics and Geology, University of Perugia, Perugia, Italy

    Silvia Caponi

  5. Biomedical Engineering Department, Ben-Gurion University of the Negev, Be’er-Sheva, Israel

    Alberto Bilenca

  6. Max Planck Institute for the Science of Light, Erlangen, Germany

    Jochen Guck

  7. Max-Planck-Zentrum für Physik und Medizin, Erlangen, Germany

    Jochen Guck

  8. Fischell Department of Bioengineering, University of Maryland, College Park, MD, USA

    Giuliano Scarcelli

  9. Maryland Biophysics Program, University of Maryland, College Park, MD, USA

    Giuliano Scarcelli

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  1. Irina Kabakova
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Contributions

Introduction (I.K. and S.C.); Experimentation (all authors); Results (all authors); Applications (I.K., J.Z., J.G. and G.S.); Reproducibility and data deposition (J.Z., S.C. and I.K.); Limitations and optimizations (J.G., G.S. and S.C.); Outlook (J.G.); Overview of the Primer (all authors).

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Correspondence to Irina Kabakova.

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Peer review information

Nature Reviews Methods Primers thanks Giuseppe Antonacci, Robert Prevedel, Seok Hyun Yun and Vladislav V. Yakovlev for their contribution to the peer review of this work.

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Related links

Bio-Brillouin Society: https://www.biobrillouin.org/

Supplementary information

Glossary

Abbe diffraction limit

This determines the size of the spot to which the light can be focused based on the law of diffraction. It is approximately proportional to half of the light wavelength, setting the lower limit to the resolution of a classical optical system.

Beat signal

The wave pattern formed via interference of two waves with slightly different frequencies.

Bragg angle

The angle between the incident light beam and the diffracting grating plane.

Brillouin frequency shift

(BFS). The frequency shift that a light wave experiences undergoing Brillouin scattering process.

Elasticity tensor

A fourth-rank tensor describing the stress–strain relation in a linear elastic material. Most materials can be described as linear elastic under small deformation approximation.

Etalon

A device consisting of two reflecting glass plates, employed for measuring the light spectrum.

Longitudinal modulus

The ratio of axial stress to axial strain under confined uniaxial deformation (the object allowed to deform along one axis only, with possible changes in volume).

Matrix pencil method

A computational technique based on diagonal matrices, particularly useful to estimate parameters of complex exponential signals buried in noise.

Mechanotransduction

The conversion of an external mechanical signal into intracellular biochemical signals.

Micromechanical properties

The elastic and viscous properties of materials at the microscale.

Rayleigh band

Refers to spectral signals at and near the frequency of the laser, including Rayleigh light scattering, quasi-elastic scattering and stray laser light.

Shear modulus

The ratio of shear stress to shear strain, serving as the measure of shear stiffness.

Voxel

A measurement of volume in a structure to be imaged.

Young’s modulus

A measure of the ratio of applied stress to axial strain under unconfined deformation (the object is free to deform in all directions while its volume is conserved).

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Kabakova, I., Zhang, J., Xiang, Y. et al. Brillouin microscopy. Nat Rev Methods Primers 4, 8 (2024). https://doi.org/10.1038/s43586-023-00286-z

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