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Abstract With the rapid development of nanotechnology, new types of fluorescent nanomaterials (FNMs) have been springing up in the past two decades. The nanometer scale endows FNMs with unique optical properties which play a critical role in their applications in bioimaging and fluorescence-dependent detections. However, since low selectivity as well as low photoluminescence efficiency of fluorescent nanomaterials hinders their applications in imaging and detection to some extent, scientists are still in search of synthesizing new FNMs with better properties. In this review, a variety of fluorescent nanoparticles are summarized including semiconductor quantum dots, carbon dots, carbon nanoparticles, carbon nanotubes, graphene-based nanomaterials, noble metal nanoparticles, silica nanoparticles, phosphors and organic frameworks. We highlight the recent advances of the latest developments in the synthesis of FNMs and their applications in the biomedical field in recent years. Furthermore, the main theories, methods, and limitations of the synthesis and applications of FNMs have been reviewed and discussed. In addition, challenges in synthesis and biomedical applications are systematically summarized as well. The future directions and perspectives of FNMs in clinical applications are also presented.