Atherosclerosis (As) is one of the important factors leading to high mortality and disability rate of cardiovascular and cerebrovascular diseases. Since its pathogenesis and heterogeneity distribution are still not clear, the “early detection, diagnosis and treatment” of As in vivo remains a great challenge. Fluorescence imaging is widely used in basic and clinical medical fields such as disease pathogenesis research, drug efficacy evaluation, and surgical navigation, because of its unique advantages such as sensitivity, noninvasive, high spatial and temporal resolution and real-time in situ, especially near-infrared fluorescence imaging with lower background interference and deeper tissue penetration, and it provides a new opportunity for the detection and treatment of the development and progression of As. Fluorescence imaging has attracted more and more attention in the research of As. This paper reviews the new progress of fluorescence imaging technology in the study of As.