Organic afterglow nanoparticles (OANs) represent a unique class of optical materials capable of sustaining luminescence after excitation cessation. Owing to their exceptional design flexibility, tunable optical properties, and favorable biosafety profiles, OAN-based afterglow imaging has emerged as an advanced modality in tumor diagnosis and therapy. These nanostructures demonstrate significant potential in guiding precision surgical interventions and real-time monitoring of tumor treatment, including photodynamic therapy, photothermal therapy, and immunotherapy. This review systematically analyzes and discusses the luminescence mechanisms of OANs under various excitation sources, with particular emphasis on recent developments in tumor detection and treatment. Additionally, we also discuss the current challenges and future perspectives of using these nanoparticles in this field.