Biological gasotransmitters (small molecules of gases) play important roles in signal transduction mechanisms and disease treatments. Although a large number of small-molecule donors have been developed, visualizing the release of small molecules remains challenging. Owing to their unique optical properties, fluorophores have been widely applied in cellular imaging and tracking. Researchers have used various fluorophores to develop small-molecule donors with fluorescent activity for visualizing the release of small molecules and their related therapies. These include fluorophores and their derivatives such as boron-dipyrromethene (BODIPY), coumarin, 1,8-naphthalimide, hemicyanine, porphyrin, rhodamine, and fluorescein. In this review, we summarize the design concepts of functional fluorescent small-molecule donors in terms of different types of fluorophores. Then, we discuss how these donors release small molecules, and the imaging modalities and biomedical applications facilitated by their fluorescent properties. With the systematic discussion of these publications, we hope to provide useful references for the development of more practical, advanced fluorescent small-molecule donors in the future.