In this study, we developed and tested a method for the fluorescence-guided resection of glioblastoma tumors in mouse models. Unlike previous models, our approach mimics clinical conditions by targeting tumors located deeper in the brain and by using the FDA-approved imaging agent 5-aminolevulinic acid. By improving the way surgery is incorporated into preclinical models, our method provides a valuable tool for testing new therapies and better understanding tumor behavior after surgery. This work could help bridge the gap between experimental research and clinical outcomes for glioblastoma patients.