Background: The middle genicular artery (MGA) is responsible for the blood supply to the cruciate ligaments and synovial tissue. Traumatic sports injuries and surgical procedures (open and arthroscopic) can cause vascular damage. Little attention has been devoted to establish safe parameters for the MGA. Purpose: To investigate the anatomy of the MGA and its relation to the posterior structures of the knees, mainly the posterior capsule and femoral condyles, and to establish safe parameters to avoid harming the MGA. Study Design: Descriptive laboratory study. Methods: Dissection of the MGA was performed in 16 fresh, unpaired adult human cadaveric knees with no macroscopic degenerative or traumatic changes and no previous surgeries. The specimens were meticulously evaluated with emphasis on preservation of the MGA. The distances from the MGA to the medial and lateral femoral condyles were measured. The Mann-Whitney test was used for statistical analysis. Results: In all specimens, the MGA emerged from the anterior aspect of the popliteal artery, distal to the superior genicular arteries, and had a short distal trajectory toward the posterior capsule where it entered proximal to the oblique popliteal ligament. The artery lay in the midportion between the condyles. The distance between the posterior aspect of the tibia and the point of entry of the MGA into the posterior joint capsule was 23.8 +/- 7.3 mm (range, 14.72-35.68 mm). There was no correlation between an individual's height and the distance of the entrance point of the MGA into the posterior joint capsule to the posterior superior corner of the tibia. Conclusion: The middle genicular artery lies in the midportion between the medial and lateral femoral condyles. Clinical Relevance: This knowledge is important for the preservation of the blood supply during posterior knee surgical procedures and to settle a secure distance between the posterior aspect of the tibia and the MGA input. This could decrease and prevent iatrogenic vascular injury risk to the MGA.