Objective To retrospectively assess the efficacy of 3D arterial spin labeling (3DASL) based on fast spin echo (FSE) technique in the preoperative grading of glioma. Methods Thirty-five patients who were suspected with glioma clinically underwent conventional MR imaging, 3D-ASL, and dynamic susceptibility contrast (DSC) imaging. The DSC and 3D ASL -derived index relative maximal tumor blood flow (rTBFmax) were generated from the tumors. Student t test was used to compare rTBFmax between 3D-ASL and DSC. Wilcoxon rank sum test was used to compare 3D-ASL and DSC-MRI indices for differences between low- and high- grade gliomas. Results The histological diagnosis of excised tumor from the 35 patients included in this study demonstrated that 18 patients had high-grade gliomas (11 had WHO grade IV, 7had WHO III), and 13 had low-grade gliomas ( Had WHO II). On Student t test, there was no significant difference between 3D-ASL and DSC -derived rTBFmax (P>0.05). There were significant differences between high- and low- grade glioma rTBFmax (P<0.01) for the two methods of 3D-ASL and DSC in high- and low- grade gliomas were respectively (2.34±1.21), (0.69±0.36), (2.55±1.06) and (0.72±0.52). Conclusion 3D-ASL can be utilized as a routine technique for evaluating the hemodynamics of gliomas. The combination of 3DASL and conventional MR imaging may improve the accuracy of preoperative grading in glioma.