Fluorescent measurement of microalgal neutral lipids

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

“…In previous studies, Nile red has been successfully used as a fluorescence probe for the detection of both neutral and polar lipids in algae from several classes (Cooksey et al, 1987;Lee et al, 1998;Elsey et al, 2007). However, this method failed to adequately detect neutral lipid content in several green algae (Fig.…”

“…A Nile red fluorescence method which has been previously successfully applied to a number of microalgae (Cooksey et al, 1987;Lee et al, 1998 andElsey et al, 2007) was evaluated for nine species of microalgae representing various classes, including C. lauderi, E. huxleyi, R. salina, Nannochloropsis sp., P. pinguis, C. cohnii, C. vulgaris, C. zofingiensis, and Pseudochlorococcum sp. After staining with Nile red, strong fluorescence signals were detected in all strains except for the three green algae strains (Fig.…”

“…Nile red, a lipid-soluble fluorescent dye, has been frequently employed to evaluate the lipid content of animal cells and microorganisms, such as mammalian cells (Genicot et al, 2005), bacteria (Izard and Limberger, 2003), yeasts (Evans et al, 1985;Kimura et al, 2004), zooplankton (Kamisaka et al, 1999), and microalgae (McGinnis et al, 1997;Eltgroth et al, 2005;Elsey et al, 2007). However, most of the studies have provided only a qualitative or semi-quantitative analysis of the lipids.…”

“…Of the algal species and strains analyzed by the Nile red method, most have belonged to the Bacillariophyceae (Cooksey et al, 1987;McGinnis et al, 1997), Xanthophyceae (Eltgroth et al, 2005), Chrysophyceae (de la Jara et al, 2003) Phaeophyceae (Brzezinski et al, 1993) with only a few species from the Chlorophyceae (Lee et al, 1998;Elsey et al, 2007). On the other hand, over the past several decades, numerous algal species and strains belonging to the Chlorophyceae have been reported to contain, based on the conventional solvent extraction and gravimetric determination analysis, high levels (25-50% of dry weight) of neutral lipid, suggesting that this class of algae may represents a large pool of organisms from which desirable candidates could be obtained for lipid/oil feedstock production (Hu et al, 2006(Hu et al, , 2008.…”

Utilization of biodiesel-derived glycerol or xylose for increased growth and lipid production by indigenous microalgae

“…Chen et al 2009;Elsey et al 2007;Huang et al 2009;Kou et al 2013;Lee 1998 work we adapted the method previously described by . After 17 days of incubation, 50µL of the algal culture sample was stained with Nile Red at 0.5µg/mL final concentration, using dimethylsulfoxide (DMSO) at 25% as carrier For the lipid profile analysis, triacylglycerols (TAGs) were extracted and transesterified in a single step, following methods already described (Cao et al 2013).…”