Although ovine stifle models are commonly used to study osteoarthritis, meniscal pathology and cruciate ligament injuries and repair, there is little information about the anatomy of the joint or techniques for synovial injections. The objectives of this study were to improve anatomical knowledge of the synovial cavities of the ovine knee and to compare intra-articular injection techniques. Synovial cavities of 24 cadaver hind limbs from 12 adult sheep were investigated by intra-articular resin, positive-contrast arthrography, computed tomography (CT) arthrography and gross anatomical dissection. Communication between femoro-patellar, medial femoro-tibial and lateral femoro-tibial compartments occurred in all cases. The knee joint should be considered as one synovial structure with three communicating compartments. Several unreported features were observed, including a communication between the medial femoro-tibial and lateral femoro-tibial compartments and a latero-caudal recess of the lateral femoro-tibial compartment. No intermeniscal ligament was identified. CT was able to define many anatomical features of the stifle, including the anatomy of the tendinous synovial recess on the lateral aspect of the proximal tibia under the combined tendon of the peroneus tertius, extensor longus digitorum and extensor digiti III proprius. An approach for intra-articular injection into this recess (the subtendinous technique) was assessed and compared with the retropatellar and paraligamentous techniques. All three injection procedures were equally successful, but the subtendinous technique appeared to be most appropriate for synoviocentesis and for injections in therapeutic research protocols with less risk of damaging the articular cartilage.