Nitroxyl (HNO) has been identified as an important signaling molecule in biological systems and plays critical roles in many physiological processes. Fluorescence imaging could provide a robust approach to explore the biological formation of HNO and its physiological functions. Herein, we summarize the organic reaction types for constructing HNO probes and specifically focus on review of the recent advances in the development of the reaction-based HNO probes and their imaging applications in living systems.