The afterglow imaging technology avoids the interference of biological tissue autofluorescence and has ultra-high imaging signal-to-noise ratio， showing great application prospects in the field of biomedical imaging. The generation and decomposition of high-energy dioxetane intermediates enable the storage and release of light energy. As a result， afterglow materials that can be oxidized by singlet oxygen to form dioxetane intermediates exhibit prolonged luminescence. The dioxetane intermediates-based afterglow luminescent materials have good biocompatibility and structural diversity， and can extend the afterglow emission range and enhance afterglow intensity through structural modification， achieving more accurate afterglow imaging. This review summarizes the luminescence mechanism and construction strategies of dioxetane-based afterglow materials， introduces the design strategies of multi-component and single-molecule afterglow materials， and discusses in detail the reported afterglow substrates and their luminescence mechanisms. Moreover， the design strategies of afterglow nanoprobes and their latest developments in disease diagnosis， biosensing， and imaging were discussed and classified. Finally， the challenges and future development prospects of this type of material in clinical translation were analyzed.

afterglow luminescence;dioxetane;afterglow material;disease diagnosis;biological imaging