Brillouin scattering microscopy is the only potential tool to realize microscopic mapping of mechanical property in multicellular system (i.e. colony, tissue) with sub-cell resolution. We built a laser-scanning Brillouin microscope system for our biological study on spatial heterogeneity of stiffness in multicellular system. High-numerical-aperture (NA) objective lens (NA ≥ 0.7) and a dual-VIPA based spectrometer were employed to achieve high spatial resolution and high sensitivity, respectively. Addition of a spatial filter at a Fourier plane of the EMCCD detector surface effectively rejected strongly reflected excitation light without loss of Brillouin scattering signal, which accordingly allowed us to observe cells just above glass substrate surface. We performed three-dimensional imaging of Brillouin scattering to see three-dimensional stiffness distribution within a multicellular system. It was found that cells in relatively central region or in close vicinity of glass substrate have higher stiffness, which agrees to biological prediction. We also found presence of anomaly cells with much higher elasticity than surrounding cells. The stiffness imaging was applied to various kinds of multicellular systems including ES cell colonies upon differentiation and artificial tissue grown from iPS cells. The results certified the effectiveness of Brillouin scattering microscope in mechanobiology, developmental biology and regenerative medicine. Several practical issues for biomedical application will also be discussed.
Taro Ichimura and Tomonobu M. Watanabe, "Brillouin scattering imaging of stiffness heterogeneity in multicellular systems (Conference Presentation)," Proc. SPIE 10496, Optical Elastography and Tissue Biomechanics V, 104960R (Presented at SPIE BiOS: January 28, 2018; Published: 15 March 2018); https://doi.org/10.1117/12.2288381.5752218882001.
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