Background: The aim of this study was to evaluate the association between prostate cancer (PCa) vascularity detected by superb microvascular imaging (SMI) and Gleason score in biopsy specimens. Material/Methods: A total of 119 patients with suspected PCa before biopsy underwent gray-scale ultrasound (US), color Doppler ultrasound (CDUS), and SMI imaging between June 2018 and March 2019. Vascularity quantity was assessed by SMI and compared with that of CDUS. The vessel parameter was also compared with the Gleason score. The sensitivity of PCa was compared between transrectal ultrasound guided systematic biopsy (SB) and SMI-guided targeted biopsy (SMI-guided TB). Results: Pathology confirmed 74 of 119 patients had PCa. The microvascular quantity of PCa patients was significantly higher than that of non-malignant patients. SMI detected blood vessels in 97.3% (72/74) in the malignant group, while CDUS identified blood flow signals in 90.5% (67/74) of the PCa group. SMI visualized enriched microvascular in PCa of Gleason 8 (54.5%) and Gleason 9 (92.3%). There was a positive correlation between microvascular quantity detected by SMI and Gleason score, with a correlation coefficient of 0.373 (P<0.001). SMI-guided TB cores were significantly more likely than SB cores to detect PCa (OR=12.83, P<0.001). Conclusions: SMI could be promising as a useful imaging technique in the detection and characterization of PCa. There was a positive correlation between microvascular quantity detected by SMI and Gleason score.