Photodynamic therapy (PDT) is a promising organ-preserving treatment for cancer. However, its widespread clinical use has long been hindered by the lack of reliable methods for intraoperative dosimetric monitoring. This review summarizes the results of experimental and clinical studies conducted in collaboration with leading medical centers in Russia on the development and implementation of fluorescence navigation methods for the personalization of PDT. The review examines the characteristics of photosensitizers (PSs) based on 5-aminolevulinic acid (5-ALA) and chlorin e6 (Ce6), causing differences in the mechanisms of accumulation, intracellular localization, and pathways of photoinduced cell death. Along with classical PSs, indocyanine green is used as an auxiliary near-infrared fluorophore. The potential of using a combination of PSs (e.g., Ce6 and 5-ALA-induced protoporphyrin IX) to achieve a synergistic effect is demonstrated. Particular attention is paid to spectroscopic navigation in the near-infrared range, which allows for real-time assessment of fluorophore photobleaching, hemoglobin oxygenation, and the optical properties of tissues, thereby enabling the adaptation of irradiation parameters to the individual characteristics of the tumor. The results of clinical trials of the developed methods in the treatment of cervical neoplasms, cholangiocellular carcinoma, basal cell carcinoma of the skin, and others are presented. The body of data demonstrates that fluorescence navigation serves as the foundation for personalizing PDT, enhancing its efficacy and safety, and expanding the method’s clinical potential.