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High-sensitivity and high-specificity biomechanical imaging by stimulated Brillouin scattering microscopy

Nature Methods volume 17, pages 913–916 (2020)Cite this article

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Abstract

Label-free, non-contact imaging with mechanical contrast and optical sectioning is a substantial challenge in microscopy. Spontaneous Brillouin scattering microscopy meets this challenge, but encounters a trade-off between acquisition speed and the specificity for biomechanical constituents with overlapping Brillouin bands. Stimulated Brillouin scattering microscopy overcomes this trade-off and enables the cross-sectional imaging of live Caenorhabditis elegans at the organ and subcellular levels, with both elasticity and viscosity contrasts at high specificity and with practical recording times.

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Fig. 1: Principle and method of SBS microscopy.
Fig. 2: Mechanical SBS imaging of live C. elegans nematodes at the organ and subcellular levels.

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Data availability

All imaging and spectral data that support the findings of this study and which are not available from public repositories owing to university constraints are available from the corresponding authors upon reasonable request and with the permission of Ben-Gurion University of the Negev.

Code availability

The SBS image reconstruction codes that support the findings of this study are available from the corresponding authors upon reasonable request and with the permission of Ben-Gurion University of the Negev under the BSD license. The code used to analyze the spectral data is provided under the BSD license as Supplementary Software along with representative sample data. The LabVIEW program for controlling the microscope, which is hardware-dependent, is available from the corresponding author upon request.

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Acknowledgements

A.B. acknowledges the support of the Israel Science Foundation (grant no. 1173/17). I.R. acknowledges the support of the Azrieli Foundation.

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Authors and Affiliations

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

    Itay Remer, Roni Shaashoua & Alberto Bilenca

  2. Agilent Research Laboratories, Petach Tikva, Israel

    Itay Remer

  3. Department of Life Sciences, Ben-Gurion University of the Negev, Be’er Sheva, Israel

    Netta Shemesh & Anat Ben-Zvi

  4. Ilse Katz Institute for Nanoscale Science and Technology, Ben-Gurion University of the Negev, Be’er-Sheva, Israel

    Alberto Bilenca

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  1. Itay Remer
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  2. Roni Shaashoua
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  4. Anat Ben-Zvi
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  5. Alberto Bilenca
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Contributions

I.R. and A.B. initiated and supervised the project; N.S and A.B.-Z. developed the C. elegans protocol. I.R., R.S. and N.S. conducted the experiments; I.R. and A.B. developed the SBS microscope and analyzed the data; I.R. and A.B. wrote the manuscript and all the coauthors contributed to the manuscript.

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Correspondence to Itay Remer or Alberto Bilenca.

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Competing interests

I.R. is an employee of Agilent Technologies. All other authors declare no competing financial interests.

Additional information

Peer review information Nina Vogt was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Supplementary Information

Supplementary Figures 1–7, Tables 1 and 2, Notes 1–4 and References

Reporting Summary

Supplementary Video 1

Cross-sectional SBS imaging of the head of the same nematode presented in Fig. 1e and under the same conditions. At each image pixel, the measured SBG spectrum was fitted to a double-Lorentzian line shape to retrieve the SBG spectral parameters of the mechanical constituent of the nematode. Seventeen z-stacks were acquired with 5-μm intervals and 0.25-NA lenses. At depths closer to the agar pads, the SBG spectrum measured in the tissue that surrounds the pharynx includes a dominant SBG component of the agar (Supplementary Note 4). This limits the accuracy with which the SBG spectral parameters of this tissue was retrieved and results in variations with depth of these parameters. Images contain 100 × 200 pixels, resulting in total image and video recording times of 400 s and 113 min, respectively. Scale bar, 20 μm.

Supplementary Software

Matlab code that analyzes spectral data along with representative Supplementary Data.

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Remer, I., Shaashoua, R., Shemesh, N. et al. High-sensitivity and high-specificity biomechanical imaging by stimulated Brillouin scattering microscopy. Nat Methods 17, 913–916 (2020). https://doi.org/10.1038/s41592-020-0882-0

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