Abstract: Targeted near-infrared fluorescence (NIRF) probes have garnered widespread attention because of their unique advantages in early tumor diagnosis, intraoperative navigation, and personalized therapy. NIRF offers strong tissue penetration, low background interference, and minimal phototoxicity, which make it an ideal tool for in vivo molecular imaging. By conjugating NIRF dyes with high-affinity targeting groups, these probes can specifically recognize tumor-associated biomarkers (such as folate receptor, prostate-specific membrane antigen, human epidermal growth factor receptor 2, epidermal growth factor receptor, carcinoembryonic antigen, etc.), which enhanced fluorescence signal intensity and contrast in tumor regions. With the continuous optimization of fluorescent dyes and upgrading of targeting groups, the applications of targeted NIRF probes in various malignant tumors are expanding, showing great potential in tumor intraoperative imaging, diagnostic imaging, and therapy. This review summarizes recent advances in targeted NIRF probes, with a focus on structural improvements, preclinical studies, and early clinical applications.