Objective This study was performed to develop and assess a genetically engineered mouse model for visualizing in vivo fluorescence of glioma cells, mural cells, and blood vessels using two-photon microscopy. Methods PDGFRβ-Cre^{+ / -}:Rosa26-tdTomato^{+ / -}genetically engineered mice underwent skull clearance and were injectedwith GL261-CFP. This was performed to study the dynamic alterations in blood vessels and mural cells during the progression and invasion of glioma using two-photon microscopy. Results PDGFRβ-Cre^{+ / -}:Rosa26-tdTomato ^{+ / -} mice were successfully bred and subjected to hematoxylin-eosin section analysis of functional organ tissues. The mice exhibited no discernible differences from C57BL/ 6 mice in terms of appearance and morphology. Cre recombinase activity was fully induced following tamoxifen treatment on day 7. Subsequent GL261-CFP inoculation demonstrated the dynamic progression of glioma proliferation and invasion, as well as vascular abnormalities and increased mural cell detachment within the tumor. Conclusions Genetically engineered mice expressing fluorescent mural cells were successfully bred. Blood vessels labeled with fluorescein isothiocyanate-dextran and blue fluorescent tumor cells were utilized. Glass discs and fixed rings were employed to replace the skulls of the mice. This allowed for the tracking of morphological and structural changes in blood vessels and vascular supporting cells following the development of brain tumors in vivo over an extended period. This model offers a valuable tool for studying brain diseases through pathological visualization.