Hypochlorite (ClO-), as a type of reactive oxygen species (ROS), plays a crucial role in the process of oxidative stress and is closely related to many diseases. Thus, developing a method for detecting and imaging of ClO- with high sensitivity and selectivity is of great significance. However, the applications of most luminescent probes are limited to the fact that the excitation and emission wavelengths of them are in the visible light region rather than in the near-infrared (NIR) region. Hence, an NIR iridium(III) complex (Mul-NIRIr) with two-photon excitation is designed for the detecting and imaging of ClO-. In the presence of ClO-, the luminescent intensity and lifetime of Mul-NIRIr are remarkably enhanced. Interestingly, Mul-NIRIr also exhibits excellent electrochemiluminescence (ECL) properties, and the ECL signal is significantly enhanced with the addition of ClO-. What is more, Mul-NIRIr is also suitable for the detection and analysis ClO- by flow cytometry. Therefore, Mul-NIRIr is developed to detect multiple signals and is successfully applied to detect exogenous and endogenous ClO- in living cells with one-photon, two-photon, and phosphorescence lifetime image microscopy (PLIM). In addition, Mul-NIRIr was successfully used for imaging of ClO- in tissues and inflammatory mouse models. All of the above results indicate that Mul-NIRIr is highly effective in detecting ClO- in living systems.