Abstract: Targeted near-infrared fluorescence (NIRF) probes have garnered widespread attention due to their unique advantages in early tumor diagnosis, intraoperative navigation, and personalized therapy. NIR fluorescence offers strong tissue penetration, low background interference, and minimal phototoxicity, making 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 FR, PSMA, HER2, EGFR, CEA, etc.), enhancing fluorescence signal intensity and contrast in tumor regions. With the continuous optimization of fluorescent dyes and upgrading of targeting groups, targeted NIRF probes are expanding their applications in various malignant tumors, showing great potential in tumor intraoperative imaging, diagnostic imaging, and therapy. This review summarizes recent advances in targeted NIRF probes, focusing on structural improvements, preclinical studies, and early clinical applications.