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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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.
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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A.B. acknowledges the support of the Israel Science Foundation (grant no. 1173/17). I.R. acknowledges the support of the Azrieli Foundation.
I.R. is an employee of Agilent Technologies. All other authors declare no competing financial interests.
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 Figures 1–7, Tables 1 and 2, Notes 1–4 and References
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.
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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