Optimization of a Nile Red method for rapid lipid determination in autotrophic, marine microalgae is species dependent

Abstract

Several studies have been conducted to develop rapid methods for quantification of lipid content in microalgae, as an alternative for time consuming gravimetric methods. Different studies showed that lipid staining with Nile Red in whole cell suspensions and subsequently quantification by the use of a spectrofluorometric device is a promising method, but a profound optimization and validation is rare. It has already been proven that the correlation curve for quantification is species dependent, but it has not yet been investigated whether this is also the case for the optimization of the Nile Red assay protocol. Therefore, two autotrophic, marine microalgae, Nannochloropsis oculata and T-Isochrysis lutea, strongly differing in e.g. cell wall structure, were selected in this study to investigate whether optimization of the Nile Red assay is species dependent. Besides this, it was checked for one of these species, Nannochloropsis, whether the lipid content, determined by the Nile Red assay, could indeed be correlated with the neutral and/or total lipid content determined by gravimetric methods. It was found that optimization of the Nile Red assay was strongly species dependent. Consequently, optimization has to be done for each species before using the assay. For Nannochloropsis, a good correlation was found between total and neutral lipid content obtained by the Nile Red assay and by gravimetric methods.

Keywords: Fluorescence; Lipid content; Lipid staining; Microalgae; Nile Red.