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

文献类型:期刊论文

Chen, wei1,2; Zhang, chengwu1; Song, lirong2; Sommerfeld, milton1; Hu, qiang1
SourceJournal of microbiological methods
2009-04-01
Volume77Issue:1Pages:41-47
KeywordBiofuel Fluorescence Green algae Microalgae Nile red Neutral lipids
ISSN0167-7012
Corresponding AuthorHu, q, arizona state univ, dept appl biol sci, 7001 e williams field rd, mesa, az 85212 usa
AbstractIsolation 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 AbstractIsolation 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.
SubjectBiochemical Research Methods; Microbiology
WOS HeadingsScience & technology ; Life sciences & biomedicine
WOS SubjectBiochemical research methods ; Microbiology
WOS Subject ExtendedBiochemistry & molecular biology ; Microbiology
WOS Keyword PlusRapid screening method ; Biotechnology ; Cells ; Dye
Indexed TypeSci
Language英语
WOS IDWos:000265314000007
Available Date2010-10-13
Citation statistics
URIhttp://www.irgrid.ac.cn/handle/1471x/58398
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
Affiliation1.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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