[Objective] To investigate the clinical application of microscope-assisted subinguinal hierarchical dissection in the treatment of varicocele (VC), so as to provide reference for clinicians. [Methods] The clinical data of 113 VC patients admitted to our hospital during Jul.2021 and Jun.2022 were retrospectively analyzed.All patients were treated with microscope-assisted hierarchical anatomical spermatic vein ligation under the external ring, with layered isolation.The number of intraoperative ligations of the internal spermatic veins and external spermatic vein, preserved arteries as well as the number of lymphatic vessels were recorded. [Results] Among the 13 patients with a mean age of (28.7±6.0) years, 98 had left-side lesion and 15 had bilateral lesion.The degree of varicocele: degree Ⅱ in 44 cases and degree Ⅲ in 69 cases.Intraoperative ligation of the internal spermatic veins: (9.18±3.21) on the left side and (9.02±3.39) on the right side.Intraoperative ligation of the external spermatic veins: (1.47±0.93) on the left side and (1.41±1.10) on the right side.Preservation of the internal spermatic arteries: (2.03±1.07) on the left side, (1.97±0.99) on the right side.Preservation of the external spermatic arteries: (1.42±0.50) on the left side and (1.40±0.50) on the right side.Preservation of lymphatics: (3.87±2.07) on the left and (3.89±1.99) on the right. [Conclusion] Hierarchical dissection ensures the integrity of the vas deferens and its vascular system, and tubular isolation aids in intraoperative reference identification, avoid omission of tiny veins and injury to lymphatic vessels, which helps to improve the surgical skills of beginners or less experienced surgeons.