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	A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae
	Chen, Wei1,2; Zhang, Chengwu1; Song, Lirong2; Sommerfeld, Milton1; Hu, Qiang1
Source	JOURNAL OF MICROBIOLOGICAL METHODS
	2009-04-01
Volume	77Issue:1Pages:41-47
Keyword	Biofuel Fluorescence Green algae Microalgae Nile red Neutral lipids
ISSN	0167-7012
Corresponding Author	Hu, Q, Arizona State Univ, Dept Appl Biol Sci, 7001 E Williams Field Rd, Mesa, AZ 85212 USA
Abstract	Isolation of high neutral lipid-containing microalgae is key to the commercial success of microalgae-based biofuel production. The Nile red fluorescence method has been successfully applied to the determination of lipids in certain microalgae, but has been unsuccessful in many others, particularly those with thick, rigid cell walls that prevent the penetration of the fluorescence dye. The conventional "one sample at a time" method was also time-consuming. In this study, the solvent dimethyl sulfoxide (DMSO) was introduced to microalgal samples as the stain carrier at an elevated temperature. The cellular neutral lipids were determined and quantified using a 96-well plate on a fluorescence spectrophotometer with an excitation wavelength of 530 nm and an emission wavelength of 575 run. An optimized procedure yielded a high correlation coefficient (R-2 = 0.998) with the lipid standard triolein and repeated measurements of replicates. Application of the improved method to several green algal strains gave very reproducible results with relative standard errors of 8.5%, 3.9% and 8.6%, 4.5% for repeatability and reproducibility at two concentration levels (2.0 mu g/mL and 20 mu g/mL), respectively. Moreover, the detection and quantification limits of the improved Nile red staining method were 0.8 mu g/mL and 2.0 mu g/mL for the neutral lipid standard triolein, respectively. The modified method and a conventional gravimetric determination method provided similar results on replicate samples. The 96-well plate-based Nile red method can be used as a high throughput technique for rapid screening of a broader spectrum of naturally-occurring and genetically-modified algal strains and mutants for high neutral lipid/oil production. (C) 2009 Published by Elsevier B.V.
English Abstract	Isolation of high neutral lipid-containing microalgae is key to the commercial success of microalgae-based biofuel production. The Nile red fluorescence method has been successfully applied to the determination of lipids in certain microalgae, but has been unsuccessful in many others, particularly those with thick, rigid cell walls that prevent the penetration of the fluorescence dye. The conventional "one sample at a time" method was also time-consuming. In this study, the solvent dimethyl sulfoxide (DMSO) was introduced to microalgal samples as the stain carrier at an elevated temperature. The cellular neutral lipids were determined and quantified using a 96-well plate on a fluorescence spectrophotometer with an excitation wavelength of 530 nm and an emission wavelength of 575 run. An optimized procedure yielded a high correlation coefficient (R(2) = 0.998) with the lipid standard triolein and repeated measurements of replicates. Application of the improved method to several green algal strains gave very reproducible results with relative standard errors of 8.5%, 3.9% and 8.6%, 4.5% for repeatability and reproducibility at two concentration levels (2.0 mu g/mL and 20 mu g/mL), respectively. Moreover, the detection and quantification limits of the improved Nile red staining method were 0.8 mu g/mL and 2.0 mu g/mL for the neutral lipid standard triolein, respectively. The modified method and a conventional gravimetric determination method provided similar results on replicate samples. The 96-well plate-based Nile red method can be used as a high throughput technique for rapid screening of a broader spectrum of naturally-occurring and genetically-modified algal strains and mutants for high neutral lipid/oil production. (C) 2009 Published by Elsevier B.V.
Subject	Biochemical Research Methods; Microbiology
WOS Headings	Science & Technology ; Life Sciences & Biomedicine
WOS Subject	Biochemical Research Methods ; Microbiology
WOS Subject Extended	Biochemistry & Molecular Biology ; Microbiology
WOS Keyword Plus	RAPID SCREENING METHOD ; BIOTECHNOLOGY ; CELLS ; DYE
Indexed Type	SCI
Language	英语
WOS ID	WOS:000265314000007
Available Date	2010-10-13
Content Type	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/7770]
Collection	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
Affiliation	1.Arizona State Univ, Dept Appl Biol Sci, Mesa, AZ 85212 USA 2.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
Recommended Citation GB/T 7714	Chen, Wei,Zhang, Chengwu,Song, Lirong,et al. A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae[J]. JOURNAL OF MICROBIOLOGICAL METHODS,2009,77(1):41-47.
APA	Chen, Wei,Zhang, Chengwu,Song, Lirong,Sommerfeld, Milton,&Hu, Qiang.(2009).A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae.JOURNAL OF MICROBIOLOGICAL METHODS,77(1),41-47.
MLA	Chen, Wei,et al."A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae".JOURNAL OF MICROBIOLOGICAL METHODS 77.1(2009):41-47.

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