Abstract
The widely applied Nile red (NR) method allows near real-time monitoring of microalgal neutral lipid accumulation. When added to a culture sample, optimally, the fluorescent dye NR penetrates the microalgal cell wall staining the intracellular neutral lipids, and the measured fluorescence is linearly correlated to the neutral lipid concentration. Here I describe an optimization protocol for determining the optimal staining parameters for each new microalgal species, followed by a basic NR staining protocol to be applied for monitoring of microalgal neutral lipid accumulation.
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References
Dutta AK, Kamada K, Ohta K (1996) Spectroscopic studies of Nile red in organic solvents and polymers. J Photochem Photobiol A Chem 93(1):57–64
Murugan NA, Rinkevicius Z, Ågren H (2011) Modeling solvatochromism of Nile red in water. Int J Quantum Chem 111:1521–1530
Lee SJ, Yoon B-D, Oh H-M (1998) Rapid method for the determination of lipid from green alga Botryococcus braunii. Biotechnol Tech 12(7):553–556
Cooksey KE, Guckert JB, Williams SA, Callis PR (1987) Fluorometric determination of the neutral lipid content of microalgal cells using Nile red. J Microbiol Methods 6:333–345
Elsey D, Jameson D, Raleigh B, Cooney MJ (2007) Fluorescent measurement of microalgal neutral lipids. J Microbiol Methods 68:639–642
Chen W, Zhang C, Song L, Sommerfeld M, Hu Q (2009) A high throughput Nile red method for quantitative measurement of neutral lipids in microalgae. J Microbiol Methods 77:41–47
Greenspan P, Fowler SD (1985) Spectrofluorometric studies of the lipid probe, Nile red. J Lipid Res 26:781–789
de la Jara A, Mendoza H, Martel A, Molina C, Nordströn L, de la Rosa V, DÃaz R (2003) Flow cytometric determination of lipid content in a marine dinoflagellate Crypthecodinium cohnii. J Appl Phycol 15:433–438
Satpati GG, Pal R (2015) Rapid detection of neutral lipid in green microalgae by flow cytometry in combination with Nile red staining—an improved technique. Ann Microbiol 65:937–949
Natunen K, Seppälä J, Koivula RJ, Pellinen J (2017) Monitoring cell-specific neutral lipid accumulation in Phaeodactylum tricornutum (Bacillariophyceae) with Nile red staining – a new method for FlowCAM. J Phycol 53:396–404
Natunen K, Seppälä J, Schwenk D, Rischer H, Spilling K, Tamminen T (2015) Nile red staining of phytoplankton neutral lipids: species-specific fluorescence kinetics in various solvents. J Appl Phycol 27:1161–1168
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917
Chen W, Sommerfeld M, Hu Q (2011) Microwave-assisted Nile red method for in vivo quantification of neutral lipids in microalgae. Bioresour Technol 102:135–141
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Natunen, K. (2017). Monitoring Microalgal Neutral Lipid Accumulation with Nile Red. In: Spilling, K. (eds) Biofuels from Algae. Methods in Molecular Biology, vol 1980. Humana, New York, NY. https://doi.org/10.1007/7651_2017_102
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DOI: https://doi.org/10.1007/7651_2017_102
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9415-1
Online ISBN: 978-1-4939-9416-8
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