Objective To evaluate the clinical value of high-resolution T2-weighted magnetic resonance imaging (HR-T₂WI), readout-segmented echo-planar imaging (RS-EPI) and HR-T₂WI plus RS-EPI in differentiating non-muscle invasive bladder cancer (NMIBC) and muscle invasive bladder cancer (MIBC). Methods A prospective study was conducted on 95 patients with bladder cancer identified by cystoscope biopsy, who underwent surgery treatment in our hospital from Jun. 2016 to Dec. 2017. All 95 patients had clear pathological staging, and received HR-T₂WI and RS-EPI examination. NMIBC and MIBC were differentiated using HR-T₂WI, RS-EPI and HR-T₂WI plus RS-EPI, and the diagnosis results were compared with the pathological results using Kappa test. The receiver operating characteristic (ROC) curve was drawn and the area under curve (AUC) of ROC was calculated to evaluate the sensitivity, specificity, accuracy of HR-T₂WI, RS-EPI and HR-T₂WI plus RSEPI. DeLong test was used to compare the AUC of three methods. Results The diagnostic results of HR-T₂WI, RS-EPI and HRT₂WI plus RS-EPI had good agreement with the pathological results (Kappa=0.770, 0.787 and 0.936; all P<0.01). The AUC, sensitivity, specificity and accuracy of HR-T₂WI, RS-EPI and HR-T₂WI plus RS-EPI were 0.899, 95.5% (42/44), 82.4% (42/51) and 88.4% (84/95), 0.891, 84.1% (37/44), 94.1% (48/51) and 89.5% (85/95), and 0.966, 93.2% (41/44), 100.0% (51/51) and 96.8% (92/95), respectively. The AUC of HR-T₂WI+RS-EPI was higher than that of HR-T₂WI and RS-EPI (Z=-2.627 8 and -2.720 5, P=0.008 6 and 0.006 5). Conclusion HR-T₂WI plus RS-EPI can be used as a preoperative non-invasive examination method to differentiate NMIBC and MIBC.