Objective To provide the anatomical basis for posterior femoral neurocutaneous vascular flap pedicled with direct popliteal artery perforator. Methods A total of 30 embalmed lower limbs of adult cadavers perfused with red latex were dissected and measured to observe the course and distribution of posterior femoral cutaneous nerve (PFCN), and the anastomoses between direct popliteal artery perforator and nutrient vessels of PFCN. Mimic operation was performed on 1 side of fresh specimen. Results PFCN started from the midpoint of the inferior gluteus maximus edge, and went down along the middle line of posterior thigh region, and the final trunk of PFCN accompanied with small saphenous vein down to the middle line of lower leg. The diameters of PFCN was (3.0 ± 0.6) mm at the inferior gluteus maximus edge, and was (2.0 ± 0.7) mm at the superior fossa poplitea. The nutrient vessels of PFCN were multi-segmental and polyphyletic. The direct popliteal artery perforator which started from popliteal artery directly was constant pierced into deep fascia about 7-11 cm above the knee joint, and its original diameter was (0.8 ± 0.2) mm. The direct popliteal artery perforator had 1-2 accompanying veins, and this perforator artery was the main nutrient vessel of the inferior segment of PFCN. The direct popliteal artery perforator gave off 5-8 small vessels which anastomosed with the 1st-3rd perforator of deep femoral artery, the obturator artery perforator, and the lateral femoral circumflex artery perforators. Then these nutrient vessels formed vascular plexus along PFCN in the middle line of posterior region of thigh. Mimic operation showed that the posterior femoral neurocutaneous vascular flap pedicled with direct poplitea artery perforator could be formed successfully. Conclusion The posterior femoral neurocutaneous vascular flap pedicled with direct popliteal artery perforator has constant blood supply and can be easily formed to repair defects around knee joint.