γ-Glutamyl Hydroxymethyl Rhodamine Green (γ-GHRG) fluorescence is a novel fluorescent agent used to enhance tissue visualization during surgical resections, especially in oncological procedures. This systematic review aims to evaluate the efficacy of γ-GHRG fluorescence in improving tumor margin detection, reducing recurrence rates, and enhancing surgical precision. A comprehensive search was conducted across PubMed, Embase, and Cochrane databases up to February 2024, following PRISMA guidelines. A total of 23 studies investigating the use of γ-GHRG fluorescence in surgical resection were included. Data on tumor visualization, surgical margin detection, and postoperative outcomes were analyzed. The included studies demonstrated that γ-GHRG fluorescence significantly improved tumor visualization in a variety of cancers, including ovarian cancer (26.1%), breast cancer (8.7%), lung cancer (8.7%), colorectal cancer (8.7%), kidney cancer (8.7%), head and neck cancer (8.7%), esophageal and gastric cancer (8.7%), prostate cancer (8.7%), brain tumors (gliomas) (4.3%), and liver cancer (4.3%). The probe's high specificity for γ-glutamyl transpeptidase (GGT), which is overexpressed in cancer cells, enabled real-time visualization of tumor margins, allowing for more precise resections. Studies also reported shorter surgical times and lower recurrence rates, particularly in high-grade tumors such as gliomas (4.3%). Although the results are promising, issues related to false positives, tissue specificity, and long-term safety were noted. γ-GHRG fluorescence shows significant potential in enhancing surgical outcomes by providing real-time guidance during tumor resections. Its high specificity for GGT and rapid fluorescence activation make it a valuable tool in cancer surgery. However, further clinical studies are required to address challenges related to sensitivity, specificity, and long-term safety, as well as to explore its application across different cancer types and surgical settings.