Nile Red, an Alternative Fluorescence Method for Quantification of Neutral Lipids in Microalgae

According to biodiesel from microalgae is an attractive fuel for several reasons such as renewable, biodegradable and environmental friendly. Thus, this study, green microalgae Scenedesmus acutus PPNK1 isolated from natural water, was selected based on high growth rates, easy cultivation and high lipid content. The Nile red fluorescence method has been successfully applied to the determination of lipids in S. acutus PPNK1. The combination of the method to the lipid composition in algal cells showed the yellow fluorescence under fluorescent microscope. Interestingly, maximum cell numbers and biomass concentration were obtained at 5.44´107 cells/mL and 1.60 g/L when it was cultivated in BG-11 medium while in case of BG-11 with nitrogen deprivation (N 0.25 g/L), accumulated lipid content in cells (44.67%) was achieved that was higher than that found in case of BG-11 medium at about 2 times (22.63%).





References:
[1] A. L. Ahmad, N. H. Mat Yasin, C. J. C. Derek,, and J. K. Lim. 2011.
Microalgae as a sustainable energy source for biodiesel production: A
review. Renewable and Sustainable Energy Reviews. 15,584- 593.
[2] T. M. Mata, A. A. Martins and N. S. Caetano 2010. Microalgae for
biodiesel production and other applications. Renewable and Sustainable
Energy Reviews. 14, 217-232.
[3] C. Y. Chen, K. L. Yeh, R. Aisyah, D. J. Lee, and J. S. Chang 2011.
Cultivation, photobioreactor design and harvesting of microalgae for
biodiesel production: A critical review Bioresource Technology. 102,
71-81.
[4] W. Chen, M. Sommerfeld, M., and Q. Hu. 2011. Microwave-assisted
Nile red method for in vivo quantification of neutral lipids in microalgae.
Bioresource Technology. 102, 135-141.
[5] S. H. Ho, W. M. Chen and J. S. Chang. 2010. Scenedesmus obliquus
CNW-N as a potential candidate for CO2 mitigation and biodiesel
production. Bioresource Technology. 101, 8725-8730.
[6] Q. Lin. and J. Lin. 2011. Effects of nitrogen source and concentration on
biomass and oil production of a Scenedesmus rubescens like microalga.
Bioresource Technology. 102, 1615-1621.
[7] J. Msanne, D. Xu, A. R. Konda, J. A. Casas-Mollano, T. Awada, E. B.
Cahoon and H. Cerutt. 2012. Metabolic and gene expression changes
triggered by nitrogen deprivation in the photoautotrophically grown
microalgae Chlamydomonas reinhardtii and Coccomyxa sp. C-169.
Phytochemistry. 75, 50-59.
[8] S. White, A. Anandraj, and F. Bux. 2011. PAM fluorometry as a tool to
assess microalgal nutrient stress and monitor cellular neutral lipids.
Bioresource Technology. 102, 1675-1682.
[9] D. Yan, Y. LU, Y. F. Chen, and Q. Wu. 2011. Waste molasses alone
displaces glucose-based medium for microalgal fermentation towards
cost-saving biodiesel production. Bioresource Technology.102, 6487-
6493.
[10] T. Govender, L. Ramanna, I. Rawat and F. Bux. 2012. BODIPY
staining, an alternative to the Nile Red fluorescence method for the
evaluation of intracellular lipids in microalgae. Bioresource Technology.
114, 507-511.
[11] E. Bertozzini, L. Galluzzi, A. Penna, and M. Magnani, 2011.
Application of the standard addition method for the absolute
quantification of neutral lipids in microalgae using Nile red. Journal of
Microbiological Methods. 87, 17-23.
[12] G. H. Huang, G. Chen and F. Chen. 2009. Rapid screening method for
lipid production in alga based on Nile red fluorescence. Biomass and
Bioenergy. 33, 1386-1392.
[13] Y. Y. Pan, S. T. Wang, L. T. Chuang, Y. W. Chang, and C. N. Nathan
Chen, 2011. Isolation of thermo-tolerant and high lipid content green
microalgae: Oil accumulation is predominantly controlled by
photosystem efficiency during stress treatments in Desmodesmus.
Bioresource Technology. 102, 10510-10517.
[14] F. Han, J. Huang, Y. Li, W. Wang, J. Wang, J. Fan, and G. Shen. 2012.
Enhancement of microalgal biomass and lipid productivities by a model
of photoautotrophic culture with heterotrophic cells as seed. Bioresource
Technology.118, 431-437.
[15] A. Converti, A. A. Casazza, E. Y. Ortiz, P. Perego, and M. D. Borghi.
2009. Effect of temperature and nitrogen concentration on the growth
and lipid content of Nannochloropsis oculata and Chlorella vulgaris for
biodiesel production. Chemical Engineering and Processing, 48, 1146-
1151.