Cranial cruciate ligament rupture is a common cause of femorotibial joint instability in the dog. Numerous techniques including several tibial osteotomies have been described for stabilization, but there is no current consensus on the best method. The instantaneous center of rotation (ICR) can aid investigations of pathological joint movement, but its use is problematic in the femorotibial joint due to combined rotation and translation during flexion and extension. Using fluoroscopic images from an earlier cadaveric study of canine joint stability, an interpolation method was used to create repeatable rotational steps across joint situations, followed by least squares approximation of the ICR. The ICR in intact joints was located mid-condyle but displaced significantly (P < 0.001) proximally following cranial cruciate ligament transection and medial meniscal release. Individual joints appear to respond differently to destabilization. Triple tibial osteotomy partially restored ICR location during early movement from flexion to extension. Joint instability significantly altered the proportions of rolling and gliding movement at the joint surface (P < 0.02), which triple tibial osteotomy partially improved. While triple tibial osteotomy restores joint stability ex vivo and clinically, normal biomechanics of the joint are not restored. The methods described here may prove useful for comparison of osteotomy techniques for stabilization of the cranial cruciate ligament deficient femorotibial joint in dogs.