Objective To investigate the advantage of neuronavigation assisted microsurgical resection of gliomas in eloquent areas of brain. Methods The clinical data of 99 patients with gliomas in eloquent areas of brain and underwent microsurgery in the First Affiliated Hospital of Sun Yat-sen University from January 2012 to December 2017 were analyzed retrospectively. All patients were divided into two groups: neuronavigator-guided microsurgery group (neuronavigation group,46 cases) and non-neuronavigator-guided microsurgery group (non-neuronavigation group,53 cases). The neuronavigation group received neuronavigation assisted microsurgery, while the non-neuronavigation group received general microsurgery. MRI examination, Karnofsky performance scale (KPS) score and symptomatic improvement rate were used to analyze the extent of resection and postoperative function. Results The total resection rate of neuronavigation group was 91.3% (42/46). Comparing with 73.6% (39/53) total resection rate in non-neuronavigation group,the difference of total resection rate was statistically significant (Z=-2.343,P＜0.05). The postoperative symptoms improved both in neuronavigation group and non-neuronavigation group,and the KPS score at discharge were higher than that before operation (P＜0.05). No aggravation of symptoms and signs or new complications occurred in all patients except one case of postoperative muscle strength decline in non-neuronavigation group after the surgery. According to tumor size,the total resection rate of neuronavigation group and non-neuronavigation group among patients whose maximum size of tumor≥3cm were 89.5%(34/38) and 72.5%(37/51) respectively. The difference of total resection rate was statistically significant between these two groups (Z=-2.040,P＜0.05). Conclusion Neuronavigation assisted microsurgical resection of gliomas in brain eloquent areas can improve the rate of total resection and postoperative quality of life without increase in postoperative complications.