AbstractLipid droplets (LDs) play a key role as the hub for lipid metabolism to maintain cellular metabolic homeostasis. Understanding the functions and changes of LDs in different pathological conditions is crucial for identifying new markers for diagnosis and discovering new targets for treatment. In recent years, coherent Raman scattering (CRS) microscopy has been popularized for the imaging and quantification of LDs in live cells. Compared with spontaneous Raman scattering microscopy, CRS microscopy offers a much higher imaging speed while maintaining similar chemical information. Due to the high lipid density, LDs usually have strong CRS signals and therefore are the most widely studied organelle in the CRS field. In this review, we discuss recent achievements using CRS to study the quantity, distribution, composition, and dynamics of LDs in various systems.