DFHBI is widely used in bioimaging and molecular biology as a fluorescent activator for RNA aptamers such as Spinach and Broccoli. When bound to these aptamers, DFHBI emits strong green fluorescence, enabling real-time visualization of RNA in live cells without the need for traditional fluorescent proteins. This makes it a valuable tool for tracking gene expression, monitoring RNA localization, and studying RNA dynamics in living systems. Its cell permeability and relatively low background fluorescence enhance its utility in cellular assays. Additionally, DFHBI is employed in the development of biosensors for detecting metabolites, ions, and enzymatic activity by coupling fluorescence activation to specific molecular interactions.