Prostate cancer, the second most common malignancy in men, frequently necessitates biopsy for accurate diagnosis. This study presents a transperineal cone-shaped double-hole prostate biopsy method guided by micro-ultrasound, performed under local anesthesia. This innovative technique offers a minimally invasive approach that maintains high diagnostic accuracy while enhancing patient comfort and reducing adverse events. The cone-shaped double-hole method is designed to minimize tissue trauma and reduce recovery times. The dual-plane ultrasound probe facilitates precise, real-time needle navigation and improves lesion-targeting accuracy. Our retrospective analysis of 526 cases demonstrated that the improved method significantly reduced the procedure time for lesion localization from 30 min to 10 min using dual-plane probe guidance. Furthermore, statistical comparisons revealed a marked reduction in pain scores, and notably, the infection rate has remained at zero to date. Compared to previous techniques that required general or spinal anesthesia, this method minimizes trauma and expedites recovery, with only two small needle punctures visible on the skin post-procedure. The findings underline the efficacy and safety of this simple, minimally invasive prostate biopsy technique under micro-ultrasound guidance, offering an alternative that improves patient outcomes without compromising diagnostic accuracy.