Low toxicity and fluorescent nanomaterials have many advantages in biological imaging. Herein, a novel and facile aqueous‐phase approach to prepare biocompatible and fluorescent Ag₂S nanoclusters (NCs) is designed and investigated. The resultant Ag₂S NCs show tunable luminescence from the visible red (624 nm) to the near infrared (NIR; 724 nm) corresponding to the increasing size of the NCs. The key for preparing tunable fluorescent Ag₂S NCs is the proper choice of capping reagent, glutathione (GSH), and the novel sulfur‐hydrazine hydrate complex as the S²⁻ source. As a naturally occurring and readily available tripeptide, GSH functions as an important scaffold to prevent NCs from growing large nanoparticles. Additionally, GSH is a small biomolecule with several functional groups, including carboxyl and amino groups, which suggests the resultant Ag₂S NCs are well‐dispersed in aqueous solution. These advantages make the as‐prepared Ag₂S NCs potentially applicable to biological labeling as well. For example, the resultant Ag₂S NCs are used as a probe for MC3T3‐EI cellular imaging.