Purpose:While MLC tracking has been successfully used for motion compensation of moving targets, current real-time target localization methods rely on correlation models with x-ray imaging or implanted electromagnetic transponders rather than direct target visualization. In contrast, ultrasound imaging yields volumetric data in real-time (4D) without ionizing radiation. We report the first results of online 4D ultrasound-guided MLC tracking in a phantom. Methods:A real-time tracking framework was installed on a 4D ultrasound station (Vivid7 dimension, GE) and used to detect a 2mm spherical lead marker inside a water tank. The volumetric frame rate was 21.3Hz (47ms). The marker was rigidly attached to a motion stage programmed to reproduce nine tumor trajectories (five prostate, four lung). The 3D marker position from ultrasound was used for real-time MLC aperture adaption. The tracking system latency was measured and compensated by prediction for lung trajectories. To measure geometric accuracy, anterior and lateral conformal fields with 10cm circular aperture were delivered for each trajectory. The tracking error was measured as the difference between marker position and MLC aperture in continuous portal imaging. For dosimetric evaluation, 358° VMAT fields were delivered to a biplanar diode array dosimeter using the same trajectories. Dose measurements with and without MLC tracking were compared to a static reference dose using a 33 mm γ-test. Results:The tracking system latency was 170ms. The mean root-mean-square tracking error was 1.01mm (0.75mm prostate, 1.33mm lung). Tracking reduced the mean γ-failure rate from 13.9% to 4.6% for prostate and from 21.8% to 0.6% for lung with high-modulation VMAT plans and from 5% (prostate) and 18% (lung) to 0% with low modulation. Conclusion:Real-time ultrasound tracking was successfully integrated with MLC tracking for the first time and showed similar accuracy and latency as other methods while holding the potential to measure target motion non-invasively. SI was supported by the Graduate School for Computing in Medicine and Life Science, German Excellence Initiative [grant DFG GSC 235/1].