Abstract: Objective：To explore the relationship between proton magnetic resonance spectroscopy (1H-MRS) and glioma cell density, CD34, and MIB-1 markers. Methods：Eighteen patients with glioma were examined by multi-voxel 1H-MRS and conventional magnetic resonance imaging before operation. Tissue biopsy was performed under neuronavigation according to 1H-MRS and conventional MRI. All specimens were stained with hematoxylin-eosin (H-E) staining and immunohistochemistry. Pearson correlation analysis was performed between 1H-MRS parameters including Cho/NAA, NAA/Cr, and Cho/Cr ratios and glioma cell density, CD34, and MIB-1. Results：From 18 patients, 89 pathological specimens were obtained via puncture. Cho/NAA ratio was positively correlated with cell density, CD34, and MIB-1, while NAA/Cr ratio was negatively correlated with cell density, CD34, and MIB-1. Cho/Cr ratio was positively correlated with cell density but not with CD34 or MIB-1. Conclusions：1H-MRS is highly correlated with glioma cell density, CD34, and MIB-1, which might provide reference for clinical diagnosis, treatment, and prognosis evaluation of glioma.