Heterometallic nanomaterials (HMNMs) display superior physicochemical properties and stability to monometallic counterparts, accompanied by wider applications in the fields of catalysis, sensing, imaging, and therapy due to synergistic effects between multi-metals in HMNMs. So far, most reviews have mainly concentrated on introduction of their preparation approaches, morphology control and applications in catalysis, assay of heavy metal ions, and antimicrobial activity. Therefore, it is very important to summarize the latest investigations of activity modulation of HMNMs and their recent applications in sensing, imaging and therapy. Taking the above into consideration, we briefly underline appealing chemical/physical properties of HMNMs chiefly tailored through the sizes, shapes, compositions, structures and surface modification. Then, we particularly emphasize their widespread applications in sensing of targets (e.g. metal ions, small molecules, proteins, nucleic acids, and cancer cells), imaging (frequently involving photoluminescence, fluorescence, Raman, electrochemiluminescence, magnetic resonance, X-ray computed tomography, photoacoustic imaging, etc.), and therapy (e.g. radiotherapy, chemotherapy, photothermal therapy, photodynamic therapy, and chemodynamic therapy). Finally, we present an outlook on their forthcoming directions. This timely review would be of great significance for attracting researchers from different disciplines in developing novel HMNMs.