Lipid droplets (LDs) are dynamic cytoplasmic organelles that function primarily in lipid storage, energy metabolism, membrane formation, and cell signaling. LDs store triacylglycerols (TAGs) and cholesterol esters. LDs are characterized by a non-polar lipid core and a phospholipid monolayer studded with proteins. In response to environmental conditions, these proteins facilitate the recruitment of accessory proteins and enzymes to the surface of the LD to metabolize TAGs for energy or to allow additional lipid deposits. The excessive accumulation of lipids without metabolism is related to non-alcoholic fatty liver disease (NAFLD). NAFLD affects about 30 million Americans, causing swelling of the liver cells and often resulting in cirrhosis of the liver. The mechanism behind the development of this disease is poorly understood. Because excess lipid storage is also related to other health conditions such as obesity and diabetes, the study of how and when LDs change in size is an important area of research. Currently, the primary techniques being used to measure the amount of lipid content within LDs are fluorescence microscopy and triglyceride assays. These methods are time consuming, subjective, and have many variables. The goal of our project is to investigate a flow cytometry-based method to quantify LDs in multiple cell lines and compare the success and improvement over both traditional measurement techniques. We have conducted preliminary studies and observed a dose-dependent increase in lipid staining by flow cytometry when hepatocytes, kidney cells, and macrophages are treated with increasing amounts of unsaturated fat. We also plan to discriminate between varying amounts of lipids within a mixed cell population. In the future, this technique can be applied to clinicians looking at human samples which may have multiple cell types present.<br/>

Independent measurements of lipids in mixed cell populations

253rd National Meeting (2017)

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