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Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy

Nature Microbiology volume 1, Article number: 16124 (2016) Cite this article

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Abstract

Understanding metabolism in live microalgae is crucial for efficient biomaterial engineering, but conventional methods fail to evaluate heterogeneous populations of motile microalgae due to the labelling requirements and limited imaging speeds. Here, we demonstrate label-free video-rate metabolite imaging of live Euglena gracilis and statistical analysis of intracellular metabolite distributions under different culture conditions. Our approach provides further insights into understanding microalgal heterogeneity, optimizing culture methods and screening mutant microalgae.

Microalgae-based biomaterials have gained a great deal of attention as food supplements, drugs, biodegradable plastics and biofuels over the past decade1–3. Among the various microalgae used for biomaterial production, Euglena gracilis, a species of unicellular flagellate protists, is attractive by virtue of its ability to produce both paramylon and lipids4. Paramylon is a type of β-1,3-glucan and is a potential drug candidate for several diseases, including HIV infection5 and colon cancer6, while lipids can be used as a biofuel alternative to fossil fuels7. To produce these biomaterials efficiently, chemically specific analysis of cell-to-cell variations in a heterogeneous population of live E. gracilis cells is essential for better understanding their biodiversity, which is more significant under environmental perturbations8. The analysis results can then help us develop and optimize the genetic transformation and culture conditions under which the cells alter their metabolism by modifying their biosynthetic pathways towards the formulation and accumulation of the metabolites.

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Figure 1: Label-free video-rate metabolite imaging of E. gracilis with a stimulated Raman scattering (SRS) microscope.
Figure 2: Statistical analysis of live E. gracilis cells under different culture conditions.

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Acknowledgements

This work was funded mainly by the ImPACT Program of the Council for Science, Technology and Innovation (Cabinet Office, Government of Japan) and partly by Advanced Photon Science Alliance and the Japan Society for the Promotion of Science (JSPS) KAKENHI (grant no. 25702026). Y.S. and K.G. are supported by JSPS and partly by Burroughs Wellcome Fund, respectively.

Author information

Author notes

  1. Yoshifumi Wakisaka and Yuta Suzuki: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, University of Tokyo, Tokyo 113-0033, Japan

    Yoshifumi Wakisaka & Keisuke Goda

  2. Department of Electrical Engineering and Information Systems, University of Tokyo, Tokyo 113-8656, Japan

    Yuta Suzuki & Yasuyuki Ozeki

  3. Research & Development Department, Euglena Co., Ltd, Tokyo 108-0014, Japan

    Osamu Iwata, Ayaka Nakashima & Kengo Suzuki

  4. Institute for Advanced Biosciences, Keio University, Tsuruoka, 997-0052, Yamagata, Japan

    Takuro Ito

  5. Graduate School of Media and Governance, Keio University, Fujisawa, 252-8520, Kanagawa, Japan

    Takuro Ito

  6. Department of Information Processing and Computer Science, Chiba University, Chiba 263-8522, Japan

    Misa Hirose, Ryota Domon, Mai Sugawara & Norimichi Tsumura

  7. Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan

    Hiroshi Watarai

  8. Department of Electrical and Electronics Engineering, Chiba University, Chiba 263-8522, Japan

    Tomoyoshi Shimobaba

  9. Department of Electrical Engineering, University of California, Los Angeles, 90095, California, USA

    Keisuke Goda

  10. Japan Science and Technology Agency, Tokyo 102-8666, Japan

    Keisuke Goda

Authors
  1. Yoshifumi Wakisaka
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  2. Yuta Suzuki
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  11. Tomoyoshi Shimobaba
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  12. Kengo Suzuki
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  13. Keisuke Goda
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  14. Yasuyuki Ozeki
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Contributions

Y.W. and Y.S. performed the experiments. O.I., A.N. and T.I. prepared the samples. Y.W., Y.S., M.H., R.D., M.S., N.T., T.S. and H.W. performed the data analysis. Y.W., Y.S., K.G. and Y.O. wrote the manuscript. K.G. conceived the concept. K.S., K.G. and Y.O. supervised the work.

Corresponding authors

Correspondence to Keisuke Goda or Yasuyuki Ozeki.

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Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary information

Supplementary Discussion, Supplementary References, Supplementary Figures 1–8, Legends for Supplementary Videos 1 and 2. (PDF 1993 kb)

Supplementary Video 1

Comparison in SRS imaging of motile E. gracilis in motion between frame rates of 27 fps (top) and 6.75 fps (bottom). (AVI 3246 kb)

Supplementary Video 1

3D metabolite imaging of E. gracilis. (MOV 2134 kb)

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Wakisaka, Y., Suzuki, Y., Iwata, O. et al. Probing the metabolic heterogeneity of live Euglena gracilis with stimulated Raman scattering microscopy. Nat Microbiol 1, 16124 (2016). https://doi.org/10.1038/nmicrobiol.2016.124

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