Dissecting lipid accumulation of microalgae Nannochloropsis oculata using fluorescent image analysis

Abstract

Cell suspension of Nannochloropsis oculata was cultured in a modified f/2 medium to study the changes of lipid content during phases of growth. The growth of cell suspension was determined by measuring the cell density and diameter under light microscope. To observe and evaluate the accumulation of lipid droplets in microalgae cells, lipid droplets were stained with Nile Red fluorescent dye then examined under fluorescence microscope and the obtained images were analyzed using Fiji ImageJ, an image processing program. The cell density increased quickly at the first 6 days of culture while cell diameter reached the highest value at the 8th day and 20th day of culture. The presence of lipid droplets in the cells could be observed from the 20th day of culture. The size of lipid droplets was gradually increased after 60 days. Treatment of depleted nitrogen for 4 days resulted an increase in the accumulation of lipid. The intracellular lipid accumulation during phases of growth of the cell suspension under nitrogen-depleted conditions was also discussed.