Hydrogen sulfide (H2S) plays an important role in mammals as a signaling molecule. Recently, abnormal H2S concentration has been associated with several pathophysiological states, such as diabetes mellitus, hypertension, Alzheimer's disease, and Parkinson's disease. As regulating H2S concentration can be a very prominent way of developing new drugs, many researchers have paid great attention to H2S research. To understand the role of H2S in pathophysiology and develop H2S-based therapies, it is necessary to measure the exact concentration of H2S within biological systems. But, H2S is volatile and can be easily oxidized. Besides, the active sites for several biological effects of H2S are inside the cell. Therefore, there is a need for the development of new methods for the accurate and reliable detection of H2S within live cells. This review provides a summary of recent developments in H2S detection methods for live cell analysis.