Sensing technology is a cutting-edge technological approach, which makes it much easier to extract information. Molecular interaction is an indispensable part in the sensing technology process, where the chemical, optical and electrical properties of the system change, and it also plays an important role in the information processing and sample detection. However, there are various problems with most of the current sensing technologies, such as low sensitivity, low selectivity, complicated operation, so they cannot be universally used in applications in various fields. By contrast, the high sensitivity of fluorescence detection technology enables the analytes in species could be detected at very low levels. More importantly, it can provide information on the temporal and spatial distribution of the analyzed species without damaging biological samples, which is of great value for studying the physiological functions of these species. This paper introduces three kinds of molecular interaction-based fluorescence sensing technologies and elaborates their principles, applications in recent years, analyzing their merits and demerits respectively, and finally, concluding predecessors’ research and prospecting the future trends of fluorescence sensing technologies. This paper lays the foundation of molecular interaction-based fluorescence sensing technologies and can provide background knowledge, which will offer help to the future research.