Microenvironmental parameters, including hypoxia, pH, polarity, viscosity and temperature, play pivotal roles in controlling the biological, physical or chemical behaviors of local molecules. Abnormal changes in these parameters would cause cellular malfunction or become a hallmark of the occurrence of severe diseases. Recently, a number of phosphorescent Ir(iii) complexes have been designed to respond to such parameters due to their attractive properties such as high photostability, long emission lifetimes, and environment-sensitive emission profiles. This review aims to provide a summary of the progress achieved in developing iridium-based probes responding to microenvironmental parameters in biological systems in recent years for diagnosis and treatment of diseases such as cancer and diabetes.