Early reports on the identification of fluorescent probes for receptors date back to mid-1970s. Fluorescent probes were initially used to visualize molecular targets in an analogous way to the use of fluorescent antibodies but with the same resolution as isotopically labelled ligands. In parallel to the rapid development of techniques, such as fluorescence correlation spectroscopy, multi-photon excitation fluorescence microscopy, fluorescence polarization and in vivo fluorescence imaging, fluorescent probes are becoming multifaceted tools in life science. The present review will focus on how the design of fluorescent ligands for receptors has evolved to meet the needs of most recent fluorescence applications.