The development of simple, efficient, and biocompatible organic luminescent molecules is of great significance to the clinical transformation of biomaterials. In recent years, purely organic thermally activated delayed fluorescence (TADF) materials with an extremely small single-triplet energy gap (𝚫E ST ) have been considered as the most promising new-generation electroluminescence emitters, which is an enormous breakthrough in organic optoelectronics. By merits of the unique photophysical properties, high structure flexibility, and reduced health risks, such metal-free TADF luminophores have attracted tremendous attention in biomedical fields, including conventional fluorescence imaging, time-resolved imaging and sensing, and photodynamic therapy. However, there is currently no systematic summary of the TADF materials for biomedical applications, which is presented in this review. Besides a brief introduction of the major developments of TADF material, the typical TADF mechanisms and fundamental principles on design strategies of TADF molecules and nanomaterials are subsequently described. Importantly, a specific emphasis is placed on the discussion of TADF materials for various biomedical applications. Finally, the authors make a forecast of the remaining challenges and future developments. This review provides insightful perspectives and clear prospects towards the rapid development of TADF materials in biomedicine, which will be highly valuable to exploit new luminescent materials.