An improved high-throughput Nile red fluorescence assay for estimating intracellular lipids in a variety of yeast species

Journal of Microbiological Methods

Volumn 91, Issue 2, 2012, Pages 321-328

  Sitepu, I R ^a,b   Ignatia, L ^a   Franz, A K ^a   Wong, D M ^a   Faulina, S A ^b   Tsui, M ^a   Kanti, A ^c   Boundy Mills, K ^a  

^a UNIVERSITY OF CALIFORNIA   (United States)

^b Forestry Research and Development Agency   (Indonesia)

^c INDONESIAN INSTITUTE OF SCIENCES   (Indonesia)

Author keywords

Biodiesel; Black microplate; Kinetic reading; Neutral lipids; Nile red; Oleaginous yeast

Indexed keywords

DIMETHYL SULFOXIDE; FLUORESCENT DYE; NILE RED; UNCLASSIFIED DRUG;

ARTICLE; BIOMASS; CELL MEMBRANE PERMEABILITY; CONTROLLED STUDY; CRYPTOCOCCUS HUMICOLA; CRYPTOCOCCUS RAMIREZGOMEZIANUS; CRYPTOCOCCUS VICTORIAE; FLUORESCENCE ANALYSIS; FUNGUS IDENTIFICATION; GRAVIMETRY; HIGH THROUGHPUT SCREENING; LIPID ANALYSIS; NONHUMAN; PHOTOSYNTHESIS; PICHIA MANSHURICA; PRIORITY JOURNAL; PROCESS OPTIMIZATION; PROTHECA ZOPFII; RHODOSPORIDIUM DIOBOVATUM; RHODOSPORIDIUM FLUVIALE; RHODOSPORIDIUM PALUDIGENUM; RHODOSPORIDIUM TORULOIDES; RHODOTORULA BABJEVAE; RHODOTORULA DAIRENSIS; RHODOTORULA GRAMINIS; RHODOTORULA MUCILAGINOSA; SACCHAROMYCES CEREVISIAE; SCHEFFERSOMYCES STIPITIS; YARROWIA LIPOLYTICA; YEAST;

FLUORESCENCE; HIGH-THROUGHPUT SCREENING ASSAYS; LIPIDS; OXAZINES; STAINING AND LABELING; YEASTS;

ALGAE; FUNGI;

EID: 84867236904     PISSN: 01677012     EISSN: 18728359     Source Type: Journal    
DOI: 10.1016/j.mimet.2012.09.001     Document Type: Article

References (23)

1. Ageitos J.J., Vallejo J.A., Veiga-Crespo P., Villa T.G. Oily yeasts as oleaginous cell factories. Appl. Microbiol. Biotechnol. 2011, 90:1219-1227.
2. Bligh E.G., Dyer W.J. A rapid method of total lipid extraction and purification. Biochem. Physiol. 1959, 37:911-917.
3. Botham P.P., Ratledge C. A biochemical explanation for lipid accumulation in Candida 107 and other oleaginous micro-organisms. J. Gen. Microbiol. 1979, 114:361-375.
4. Chen W., Zhang C., Song L., Sommerfeld M., Hu Q. A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae. J. Microbiol. Methods 2009, 77:41-47.
5. Cheng Y.S., Zheng Y., VanderGheynst J.S. Rapid quantitative analysis of lipids using colorimetric method in a microplate format. Lipids 2011, 46:95-103.
6. Cole T.A. Use of Nile red as a rapid measure of lipid contents in ciliates. Eur. J. Protistol. 1990, 25:361-368.
7. Elle I.C., Olsen L.C.B., Pultz D., Rødkær S.V., Færgeman N.J. Something worth dyeing for: molecular tools for the dissection of lipid metabolism in Caenorhabditis elegans. FEBS Lett. 2010, 584:2183-2193.
8. Evans C.T., Ratledge C., Gilbert S.C. A rapid screening method for lipid-accumulating yeast using a replica-printing technique. J. Microbiol. Methods 1985, 4:203-210.
9. Folch J., Lees M., Sloane-Stanley G.H. A simple method for the isolation and purification of the total lipids from animal tissues. J. Biol. Chem. 1956, 226:497-509.
10. Fowler S.D., Brown W.J., Warfel J., Greenspan P. Use of nile red for the rapid in situ quantitation of lipids on thin-layer chromatograms. J. Lipid Res. 1987, 28:1225-1232.
11. Govender T., Ramanna L., Rawat I., Bux F. BODIPY staining, an alternative to the Nile Red fluorescence method for the evaluation of intracellular lipids in microalgae. Bioresour. Technol. 2012, 114:507-511.
12. Greenspan P., Fowler S.D. Spectrofluorometric studies of the lipid probe, nile red. J. Lipid Res. 1985, 26:781-789.
13. Greenspan P., Mayer E.P., Fowler S.D. Nile red: a selective fluorescent stain for intracellular lipid droplets. J. Cell Biol. 1985, 100:955-977.
14. Hu Q., Sommerfeld M., Jarvis E., Ghirardi M., Posewitz M., Seibert M., Darzins A. Microalgal triacylglycerols as feedstocks for biofuel production: perspectives and advances. Plant J. 2008, 54:621-639.
15. Huang G.H., Chen G., Chen F. Rapid screening method for lipid production in alga based on Nile red fluorescence. Biomass Bioenergy 2009, 33:1386-1392.
16. Kimura K., Yamaoka M., Kamisaka Y. Rapid estimation of lipids in oleaginous fungi and yeasts using Nile red fluorescence. J. Microbiol. Methods 2004, 56:331-338.
17. Listenberger L.L., Brown D.A. Fluorescent detection of lipid droplets and associated proteins. Curr. Protoc. Cell Biol. 2007, 24:24.2.1-24.2.11.
18. Ratledge C. Microorganisms for Lipids. Acta Biotechnol. 1991, 5:429-438.
19. Sackett D.L., Wolff J. Nile red as a polarity-sensitive fluorescent probe of hydrophobic protein surfaces. Anal. Biochem. 1987, 167:228-234.
20. Safranyos R.G.A., Caveney S. Rates of diffusion of fluorescent molecules via cell-to-cell membrane channels in a developing tissue. J. Cell Biol. 1985, 100:736-747.
21. Shahidi F. Extraction and measurement of total lipids. Curr. Protoc. Food Anal. Chem. 2001, D1:D1.1.1-D1.1.11.
22. Suutari M., Priha P., Laakso S. Temperature shifts in regulation of lipids accumulated by Lipomyces starkeyi. J. Am. Oil Chem. Soc. 1993, 70:891-894.
23. U.S. Environmental Protection Agency Alternative fuels: Biodiesel Report #EPA420-F-06-044 (2006) Available at:, (accessed May 2012). http://www.epa.gov/smartway.