Lipid Droplet Staining in Microalgae

For LD staining in L. incisa cells 10 ml of samples were collected at selected time points of nitrogen deprivation, pelleted by centrifugation at 1000 g for 5 min and fixed with 4% (w/v) paraformaldehyde in 0.1 M PBS, pH 7.4, overnight at 4 °C. Fixed samples were washed three times with the same buffer and stained with 10 μg/ml Bodipy 493/504 (Thermo Fisher Scientific) diluted in 0.1 M PBS buffer, pH 7.4, for 1 h at RT with gentle agitation. Then samples were washed 3 times with the PBS buffer only and re-suspended in Prolong gold anti-fade reagent (Thermo Fisher Scientific). LD staining in yeast cells was performed as described in [40 (link)].