Imaging flow cytometry (IFC) is an emerging technology that combines the high-throughput analysis capabilities of traditional flow cytometry with high-resolution and high-specificity imaging of microscopes. Traditional flow cytometry is mainly used for rapid quantitative analysis of molecules at the single cell or subcellular level. The results are usually presented in the form of scatter plots, but lack information about cell morphology, structure, and subcellular signal distribution, which limits its application in cell structure research and intracellular molecular distribution analysis. IFC can image cells and has an extended depth of field function, allowing light from different focal planes to be focused on the detection plane at the same time. Therefore, through multiple detection channels, IFC can perform multi-parameter quantitative analysis of cell images, which not only retains the characteristics of traditional flow cytometry but also realizes high-resolution imaging and visualization of single cells. This article reviews the basic principles and latest progress of IFC from the perspective of two key technologies: optical imaging and image-guided cell sorting. It focuses on how this technology combines the high-throughput advantages of traditional flow cytometry with the high-resolution capabilities of microscopic imaging to overcome the limitations of traditional flow cytometry in cell morphology analysis and molecular distribution research, providing a reference for subsequent research.