Production and high throughput quantification of fucoxanthin and lipids in Tisochrysis lutea using single-cell fluorescence

Abstract

This work aimed to investigate the accumulation of fucoxanthin and lipids in Tisochrysis lutea during growth (N+) and nitrogen-starvation (N−) and to correlate these products with single-cell emissions using fluorescence-activated cell sorting (FACS). Fucoxanthin content decreased 52.94% from N+ to N− in batch cultivation; increased 40.53% as dilution rate changed from 0.16 to 0.55 d−1 in continuous cultivation. Total lipids (N−) were constant (~250 mg/g), but the abundance of neutral lipids increased from 4.87% to 40.63%. Nile red can stain both polar and neutral lipids. However, in vivo, this differentiation is limited due to an overlapping of signals between 600 and 660 nm, caused by neutral lipids concentrations above 3.48% (W/W). Chlorophyll autofluorescence (720 nm) was reported for the first time as a proxy for fucoxanthin (R2 = 0.90) and polar lipids (R2 = 0.98). FACS can be used in high throughput quantification of pigments and lipids and to select and sort cells with high-fucoxanthin/lipids.