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Label-free biomechanical imaging approaches include a variety of techniques, such as atomic-force microscopy and optical coherence elastography , that probe biomechanics at high spatial resolution in complementary regimes, but require application of an external force on the sample. Second-harmonic generation microscopy circumvents this necessity, yet its applicability is limited to a small number of structural proteins. An emerging technique, spontaneous Brillouin scattering microscopy4–15, enables label-free and non-contact biomechanical imaging in three dimensions by measuring the so-called Brillouin shift (ΩB) and linewidth (ΓB), which correspond to the frequency shift and linewidth of light that is backscattered inelastically from gigahertz-frequency longitudinal acoustic phonons that are characteristic of the mechanical components of the material.