Fluorescent probes have been used widely in bioimaging, including biological substance detection, cell imaging, in vivo biochemical reaction process tracking, and disease biomarker monitoring, and have gradually occupied an indispensable position. Compared with traditional biological imaging technologies, such as positron emission tomography (PET) and nuclear magnetic resonance imaging (MRI), the attractive advantages of fluorescent probes, such as real-time imaging, in-depth visualization, and less damage to biological samples, have made them increasingly popular. Among them, ultraviolet–visible (UV–vis) fluorescent probes still occupy the mainstream in the field of fluorescent probes due to the advantages of available structure, simple synthesis, strong versatility, and wide application. In recent years, fluorescent probes have become an indispensable tool for bioimaging and have greatly promoted the development of diagnostics. In this review, we focus on the structure, design strategies, advantages, representative probes and latest discoveries in application fields of UV–visible fluorescent probes developed in the past 3–5 years based on several fluorophores. We look forward to future development trends of fluorescent probes from the perspective of bioimaging and diagnostics. This comprehensive review may facilitate the development of more powerful fluorescent sensors for broad and exciting applications in the future.