Objective To investigate the dissection of antecubital perforator flap and the design of donor site and its clinical application. Methods The origin, trending, branches, outer diameter, distribution and anastomosis of blood vessels and nerves were studied by perfusing red, yellow and blue emulsion into 10 cadaver specimens. The area of the antecubital perforating flap was measured after injecting ink to one fresh cadaver specimen. Then, antecubital perforator flap was designed according to the source of blood supply of antecubital perforating flap and its relationship with the blood supply of forearm fascia. The soft tissue defect of elbow skin was repaired by proximal pedicle of antecubital perforating flap, and the palmar and dorsal defect in middle and lower segment of the forearm were repaired by distal pedicle of antecubitai perforating flap in seven patients. Results All the flaps were survived, with wound healing by first intention. All patients were followed up for 3-15 months, with fine appearance and good texture and color of the flaps. Conclusions Antecubital perforator flap has the advantages of constant anatomical location, perfect blood supply, simple and safe application during operation, and it is excellent in repairing the skin defect of the elbow and middle or lower segment of the forearm.