A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae

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	A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae
	Chen, Wei 1,2; Zhang, Chengwu 1; Song, Lirong 2; Sommerfeld, Milton 1; Hu, Qiang 1
刊名	JOURNAL OF MICROBIOLOGICAL METHODS
	2009-04-01
卷号	77 期号:1 页码:41-47
关键词	Biofuel Fluorescence Green algae Microalgae Nile red Neutral lipids
ISSN号	0167-7012
通讯作者	Hu, Q, Arizona State Univ, Dept Appl Biol Sci, 7001 E Williams Field Rd, Mesa, AZ 85212 USA
中文摘要	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.
英文摘要	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.
学科主题	Biochemical Research Methods; Microbiology
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemical Research Methods ; Microbiology
研究领域[WOS]	Biochemistry & Molecular Biology ; Microbiology
关键词[WOS]	RAPID SCREENING METHOD ; BIOTECHNOLOGY ; CELLS ; DYE
收录类别	SCI
语种	英语
WOS记录号	WOS:000265314000007
公开日期	2010-10-13
内容类型	期刊论文
源URL	[http://ir.ihb.ac.cn/handle/152342/7770]
专题	水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文
作者单位	1.Arizona State Univ, Dept Appl Biol Sci, Mesa, AZ 85212 USA 2.Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
推荐引用方式 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.