The measurement of five degrees-of-freedom (5-DOF) error motions, including radial, axial, and tilt motions, is crucial for ultra-precision rotary axes, which are key components of ultra-precision machine tools and instrumentation. In this study, we propose an interference-enhanced micro-vision technique to concurrently derive the 5-DOF error motions from a single-shot two-dimensional image, which was captured by a standard industrial camera equipped with an interference objective lens. By consolidating the essential features into a single optical path, the interference-enhanced micro-vision technique ingeniously merges machine micro-vision and modified white-light interference to detect in-plane and out-of-plane motions. Numerical simulations demonstrated, the basic principle for deriving the 5-DOF error motions, and the magnification of objective lens had inconsistent effects on the measurement accuracy for the radial and tilt motions, i.e. higher magnification led to higher radial accuracy but lower tilt accuracy. As practical application, the error motion detection capability was demonstrated by simultaneously measuring the 5-DOF synchronous and asynchronous error motions for a typical air bearing spindle at rotation speeds of 8.33, 108.33, and 308.33 rpm. The synchronous errors were nearly identical at various spindle speeds. However, because of system dynamics, increased vibrations were observed to be superimposed on the basic tilt error motions as the spindle speeds increased, which were verified by the vibration marks imprinted on the turned surfaces. For the 5-DOF motion measurements, the least-square fitting using large-volume edge and greyscale data of the captured image enabled super-high resolutions, despite using a camera with a relatively large pixel size and low bit depth. These results demonstrate that the proposed interference-enhanced micro-vision technique is a simple and effective tool for measuring spatial error motions in ultra-precision rotary axes.