Microbes, especially microalgae, have recently been of great interest for developing novel biofuels, drugs, and biomaterials. Imaging-based screening of live cells can provide high selectivity and is attractive for efficient bio-production from microalgae. Although conventional cellular screening techniques use cell labeling, labeling of microbes is still under development and can interfere with their cellular functions. Furthermore, since live microbes move and change their shapes rapidly, a high-speed imaging technique is required to suppress motion artifacts. Stimulated Raman scattering (SRS) microscopy allows for label-free and high-speed spectral imaging, which helps us visualize chemical components inside biological cells and tissues. Here we demonstrate high-speed SRS imaging, with temporal resolution of 0.14 seconds, of intracellular distributions of lipid, polysaccharide, and chlorophyll concentrations in rapidly moving Euglena gracilis, a unicellular phytoflagellate. Furthermore, we show that our method allows us to analyze the amount of chemical components inside each living cell. Our results indicate that SRS imaging may be applied to label-free screening of living microbes based on chemical information.
Yoshifumi Wakisaka, Yuta Suzuki, Kyoya Tokunaga, Misa Hirose, Ryota Domon, Rina Akaho, Mai Kuroshima, Norimichi Tsumura, Tomoyoshi Shimobaba, Osamu Iwata, Kengo Suzuki, Ayaka Nakashima, Keisuke Goda, and Yasuyuki Ozeki, "Label-free chemical imaging of live Euglena gracilis by high-speed SRS spectral microscopy
(Conference Presentation)," Proc. SPIE 9720, High-Speed Biomedical Imaging and Spectroscopy: Toward Big Data Instrumentation and Management, 97200H (Presented at SPIE BiOS: February 14, 2016; Published: 28 June 2016); https://doi.org/10.1117/12.2211991.4848767887001.
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