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

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 CO2 into lipid droplets (LDs). Quantification of LDs in microalgae is required for developing and optimizing algal bi...

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Main Authors: Noo Li Jeon, Sanghee Lee, Seung Bum Park, Youngjun Lee, Sang Cheol Na, Jae Woo Park
Format: Article
Language:English
Published: MDPI AG 2013-10-01
Series:Energies
Subjects:
Online Access:http://www.mdpi.com/1996-1073/6/11/5703
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author Noo Li Jeon
Sanghee Lee
Seung Bum Park
Youngjun Lee
Sang Cheol Na
Jae Woo Park
author_facet Noo Li Jeon
Sanghee Lee
Seung Bum Park
Youngjun Lee
Sang Cheol Na
Jae Woo Park
author_sort Noo Li Jeon
collection DOAJ
description 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 CO2 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.
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spelling doaj.art-c8278a4ec81b4bcea85856202d7017f12022-12-22T04:21:00ZengMDPI AGEnergies1996-10732013-10-016115703571610.3390/en6115703Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-FluorNoo Li JeonSanghee LeeSeung Bum ParkYoungjun LeeSang Cheol NaJae Woo ParkAlternative 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 CO2 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.http://www.mdpi.com/1996-1073/6/11/5703microalgaeChlamydomonas reinhardtiiSF44lipid dropletsbiodiesel
spellingShingle Noo Li Jeon
Sanghee Lee
Seung Bum Park
Youngjun Lee
Sang Cheol Na
Jae Woo Park
Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
Energies
microalgae
Chlamydomonas reinhardtii
SF44
lipid droplets
biodiesel
title Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
title_full Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
title_fullStr Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
title_full_unstemmed Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
title_short Measurement of Lipid Droplet Accumulation Kinetics in Chlamydomonas reinhardtii Using Seoul-Fluor
title_sort measurement of lipid droplet accumulation kinetics in chlamydomonas reinhardtii using seoul fluor
topic microalgae
Chlamydomonas reinhardtii
SF44
lipid droplets
biodiesel
url http://www.mdpi.com/1996-1073/6/11/5703
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