Evaluation of Lipid Droplet Size and Fusion in Bovine Hepatic Cells

The present protocol describes how to use oil red O to dye lipid droplets (LDs), calculate the size and number of LDs in a fatty acid-induced fatty hepatocyte model, and use BODIPY 493/503 to observe the process of small LDs fusing into large LDs by live cell imaging.

Our research focus on the lipid droplet number and the size induced by linoleic acid in liver cells. Additionally, we aim to provide insights into the mechanism of the fatty liver. Recent advancements in this field of research, suggests that choline, CCT alpha, established induced autophagy, can regulate lipid droplet size in bovine hepatic cells.

Moreover, living cell workstation observations have shown that phospholipids have a considerable effect on lipid droplet fusion. Currently, gene aiding, fluorescence tweezer, liquid chromatography, omics analysis, single chromatography, flow cytometry, and other technologies are being used to group the vast research in this field. The experimental method has some limitations.

For example, the phenomenon of limited fields can only be observed for lipid droplet fields and under a certain flight of waste. Oil Red O staining is more convenient, less expensive, and more widely used to measure the apparent size of lipid droplets than lipid droplet fluorescence staining. It is simple to operate and requires less equipment.

Furthermore, we directly observe this phenomenal of lipid fusion through the living cell station. This study provide a simple and repeatable method to observe lipid droplet size and fusion, which can be widely applied to lipid droplet analysis in cellular lipid metabolism research. We will continue to conduct intensive research on lipid droplets, focusing on the mechanism of lipid droplet fusion and the structure changes of fatty liver in dairy cows.