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Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor

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  • Jae Woo Park

    (Division of WCU (World Class University) Multiscale Mechanical Design, School of Mechanical Engineering, Seoul National University, Seoul 151-744, Korea
    These authors contributed equally to this work.)

  • Sang Cheol Na

    (Division of WCU (World Class University) Multiscale Mechanical Design, School of Mechanical Engineering, Seoul National University, Seoul 151-744, Korea
    These authors contributed equally to this work.)

  • Youngjun Lee

    (Department of Chemistry, Seoul National University, Seoul 151-744, Korea)

  • Sanghee Lee

    (Department of Chemistry, Seoul National University, Seoul 151-744, Korea)

  • Seung Bum Park

    (Department of Chemistry, Seoul National University, Seoul 151-744, Korea)

  • Noo Li Jeon

    (Division of WCU (World Class University) Multiscale Mechanical Design, School of Mechanical Engineering, Seoul National University, Seoul 151-744, Korea
    School of Mechanical and Aerospace Engineering, Seoul National University, Seoul 151-744, Korea)

Abstract

Alternative energy resources have become an important issue due to the limited stocks of petroleum-based fuel. Microalgae, a source of renewable biodiesel, use solar light to convert CO 2 into lipid droplets (LDs). Quantification of LDs in microalgae is required for developing and optimizing algal bioprocess engineering. However, conventional quantification methods are both time and labor-intensive and difficult to apply in high-throughput screening systems. LDs in plant and mammalian cells can be visualized by staining with various fluorescence probes such as the Nile Red, BODIPY, and Seoul-Fluor (SF) series. This report describes the optimization of LD staining in Chlamydomonas reinhardtii with SF probes via systematic variations of dye concentration, staining time, temperature, and pH. A protocol for quantitative measurement of accumulation kinetics of LDs in C. reinhardtii was developed using a spectrofluorimeter and the accuracy of LD size measurement was confirmed by transmission electron microscopy (TEM). Our results indicate that our spectrofluorimeter-based measurement approach can monitor kinetics of intracellular LDs (in control and nitrogen-source-starved Chlamydomonas reinhardtii ) accumulation that has not been possible in the case of conventional imaging-based methods. Our results presented here confirmed that an SF44 can be a powerful tool for in situ monitoring and tracking of intracellular LDs formation.

Suggested Citation

  • Jae Woo Park & Sang Cheol Na & Youngjun Lee & Sanghee Lee & Seung Bum Park & Noo Li Jeon, 2013. "Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor," Energies, MDPI, vol. 6(11), pages 1-14, October.
    • Handle: RePEc:gam:jeners:v:6:y:2013:i:11:p:5703-5716:d:30062
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    References listed on IDEAS

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    1. Zhegang Huang & Hyojin Lee & Eunji Lee & Seong-Kyun Kang & Jwa-Min Nam & Myongsoo Lee, 2011. "Responsive nematic gels from the self-assembly of aqueous nanofibres," Nature Communications, Nature, vol. 2(1), pages 1-5, September.
    2. Demirbas, Ayhan, 2007. "Importance of biodiesel as transportation fuel," Energy Policy, Elsevier, vol. 35(9), pages 4661-4670, September.
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