Materials C nbsp vulgaris culture was purchased from Carolina Biological
C. vulgaris culture was purchased from Carolina Biological Supply Company (Burlington, NC). Chemicals and reagents used in the research were obtained from Fischer Scientific (Pittsburgh, PA) and Sigma–Aldrich Co. (St. Louis, Mo).
2.2.1. Microalgae growth medium
A growth medium designated as ATCC® Medium 5 (Manassas, VA) was used. The medium was prepared by dissolving 1.0 g yeast extract, 1.0 g beef extract, 0.004 g FeSO4.7H2O, and 10 g of dextrose in 3 L DI water. The medium was autoclaved at 121 °C for 20 min, and the Miltefosine was adjusted to 7.5 prior to autoclaving for a final pH of 7.2.
2.2.2. Growing and harvesting microalgae for experiments
Optical density, measured at 600 nm, was used as an estimate of cell mass during the course of culture growth, Fig. S2 in supplementary material. At the end of the exponential growth phase, the culture was harvested and centrifuged – for 20 min at 13,000 g, which is equivalent to 9000 rpm – using Thermo Scientific Sorvall RC 6 + centrifuge (Waltham, MA. The process is not expected to damage cells because the centrifugal force is lower than that which would have an impact on cell viability .