Lipid droplets (LDs) are ubiquitous subcellular organelles for the storage of neutral lipids in cells.LDs exert multiple functions even in highly specialized mature adipocytes, including the maintenance of lipid metabolism equilibrium, the regulation of the distribution and degradation of many structural and signal proteins, and the trafficking orchestration of the cellular membrane system that is critical fora variety of important biological processes.Due to the nature of LD dynamics and the technical difficulties or limitations, the studies on the formation and maturation of LDs advanced slowly to date.Recently, the emerging technologies of real-time live cell imaging and high-content digital image analysis have provided a novel approach for the breakthroughs in this field.