Background: Glioma is the most common primary brain neoplasm. Surgical excision is the primary management modality for high-grade glioma. The aim of surgery is maximum safe resection for improving survival. Objective: The aim of the current study was to evaluate the value of intra-operative sodium fluorescein in detecting boundaries and maximizing surgical resection of glioma. Patients and methods: The study included 40 patients with pre-operative MRI showing contrast-enhanced lesions suspected to be high-grade glioma. Patients were operated on after IV injection of sodium fluorescein (5-8mg/kg) after induction of anesthesia. With the help of FL560 filters integrated into the Leica Provido microscope, all fluorescent areas could be resected. Volumetric assessment of the resected tumor tissue was calculated from pre-operative and early post-operative MRI with contrast. Results: Gross total resection was achieved in most patients (95%), while in 5% of patients we just did near-total resection. The extent of resection ranged from 96 to 99% with a mean of 98.08%. The sensitivity and specificity of fluorescein for the high-grade glioma (contrast-enhanced areas) is 95% and 100% respectively. Conclusion: The integration of intra-operative IV sodium fluorescein provided a safe, easy, and effective utility for differentiating tumorous from normal tissue thus maximizing the extent of resection of high-grade gliomas.