procedure
The experiment was designed to focus on the area of lipid droplets induced in HeLa cells treated with lauric acid. The positive control was oleic acid, which has been previously tested on HeLa cells and is proven to increase lipid droplet area. Culture plates containing about 10,000 HeLa cells were exposed to 400 micromolar of either oleic or lauric acid and left to incubate for 24 hours. To prepare the acids, they were diluted in 50% ethanol to a final concentration of 21 micromolar. BSA was dissolved in sterile filtered water to 3.5 micromolar. On the day of the experiment, BSA and the acid were mixed in a 6 to 1 ratio. In series and following the manufacturers' instructions, lipid droplets were stained green (LipidSpot 488 Lipid Droplet Stain, Biotium #70065), membranes were stained red (CF594 WGA, Biotium # 29023-1), and DAPI stain (NucBlue Live Cell Stain, Invitrogen #R37605) was used to stain the nuclei blue. Each plate was then analyzed for lipid droplet area, and data was recorded using ImageJ software. The effects of the lauric and oleic acid were compared to a negative control plate containing only ethanol. Data gathered from observations consisted of whether lipid droplet area was successfully increased. Lipid droplet area was quantified by measuring the lipid droplets of three cells from each image taken of the three groups. Data was collected and formulated into a bar graph.