High-speed X-ray video demonstrates significant skin movement errors with standard optical kinematics during rat locomotion

The sophistication of current rodent injury and disease models outpaces that of the most commonly used behavioral assays. The first objective of this study was to measure rat locomotion using high-speed X-ray video to establish an accurate baseline for rat hindlimb kinematics. The second objective was to quantify the kinematics errors due to skin movement artefacts by simultaneously recording and comparing hindlimb kinematics derived from skin markers and from direct visualization of skeletal landmarks. Joint angle calculations from skin-derived kinematics yielded errors as high as 39° in the knee and 31° in the hip around paw contact with respect to the X-ray data. Triangulation of knee position from the ankle and hip skin markers provided closer, albeit still inaccurate, approximations of bone-derived, X-ray kinematics. We found that soft tissue movement errors are the result of multiple factors, the most impressive of which is overall limb posture. Treadmill speed had surprisingly little effect on kinematics errors. These findings illustrate the significance and context of skin movement error in rodent kinematics.