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学科主题: Biochemical Research Methods; Microbiology
题名: A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae
作者: Chen, Wei1, 2; Zhang, Chengwu1; Song, Lirong2; Sommerfeld, Milton1; Hu, Qiang1
通讯作者: Hu, Q, Arizona State Univ, Dept Appl Biol Sci, 7001 E Williams Field Rd, Mesa, AZ 85212 USA
关键词: Biofuel ; Fluorescence ; Green algae ; Microalgae ; Nile red ; Neutral lipids
刊名: JOURNAL OF MICROBIOLOGICAL METHODS
发表日期: 2009-04-01
DOI: 10.1016/j.mimet.2009.01.001
卷: 77, 期:1, 页:41-47
收录类别: SCI
文章类型: Article
部门归属: [Chen, Wei; Zhang, Chengwu; Sommerfeld, Milton; Hu, Qiang] Arizona State Univ, Dept Appl Biol Sci, Mesa, AZ 85212 USA; [Chen, Wei; Song, Lirong] Chinese Acad Sci, Inst Hydrobiol, Wuhan 430072, Hubei, Peoples R China
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
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.
关键词[WOS]: RAPID SCREENING METHOD ; BIOTECHNOLOGY ; CELLS ; DYE
语种: 英语
WOS记录号: WOS:000265314000007
ISSN号: 0167-7012
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/152342/7770
Appears in Collections:中科院水生所知识产出(2009年前)_期刊论文

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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

Recommended Citation:
Chen, Wei; Zhang, Chengwu; Song, Lirong; Sommerfeld, Milton; Hu, Qiang.A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae,JOURNAL OF MICROBIOLOGICAL METHODS,2009,77(1):41-47
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