ABSTRACT Compared to fluorescence imaging, chemiluminescence imaging does not need external excitation light, and hence presents high imaging depth and signal‐to‐noise ratio without autofluorescence and phototoxicity, making it a promising tool for biological detection and analysis. However, the target‐specific activatable near‐infrared emission chemiluminescent probes still need to be developed for the precise diagnosis of diseases. In this paper, we synthesized four direct near‐infrared emission Schaap's chemiluminophores (AINCL, AIFCL, ABTCL, and APYCL) by incorporating different electronic acceptors, respectively, and studied the effect of the acceptors on the optical properties of the chemiluminophores. To achieve the specific detection of hydrogen sulfide (H 2 S)‐related diseases, we used H 2 S‐cleavable 2,4‐dinitrophenylsulfonate to cage the phenol groups in the chemiluminophores. It was demonstrated that the endogenous H 2 S in inflammations and tumors could activate effectively the chemiluminescence with high specificity, which provided the precise location of nidus in chemiluminescence imaging and allowed us to perform surgical resection.