The purpose of this work was to investigate the use of an experimental CMOS (Complementary Metal-OxideSemiconductor) APS (Active Pixel Sensor) for tracking of moving fiducial markers during radiotherapy. The APS has an active area of 5.4 x 5.4 cm and maximum full frame read-out rate of 20 frame s −1 , with the option to read out a Region-Of-Interest (ROI) at an increased rate. It was coupled to a 4 mm thick ZnWO4 scintillator which provided a quantum efficiency (QE) of 8% for a 6 MV x-ray treatment beam. The APS was compared with a standard iViewGT flat panel amorphous Silicon (a-Si) Electronic Portal Imaging Device (EPID), with a QE of 0.34% and a frame-rate of 2.5 frame s −1 . To investigate the ability of the two systems to image markers, four gold cylinders of length 8 mm and diameter 0.8, 1.2, 1.6 and 2 mm were placed on a motion platform. Images of the stationary markers were acquired using the APS at a frame-rate of 20 frame s −1 , and a dose-rate of 143 MU min −1 to avoid saturation. EPID images were acquired at the maximum frame-rate of 2.5 frame s −1 , and a reduced dose-rate of 19 MU min −1 to provide a similar dose per frame to the APS. Signal-to-Noise Ratio (SN R) of the background signal and Contrast-to-Noise Ratio (CN R) of the marker signal relative to the background were evaluated for both imagers at doses of 0.125 to 2 MU. Image quality and marker visibility was found to be greater in the APS with SN R ∼5 times greater than in the EPID and CN R up to an order of magnitude greater for all four markers. To investigate the ability to image and track moving markers the motion platform was moved to simulate a breathing cycle with period 6 s, amplitude 20 mm and maximum speed 13.2 mm s −1 . At the minimum integration time of 50 ms a tracking algorithm applied to the APS data found all four markers with a success rate of ≥92% and positional error ≤90 µm. At an integration time of 400 ms the smallest marker became difficult to detect when moving. The detection of moving markers using the a-Si EPID was difficult even at the maximum dose-rate of 592 MU min −1 due to the lower QE and longer integration time of 400 ms. This work demonstrates that a fast read-out, high QE APS may be useful in the tracking of moving fiducial markers during radiotherapy. Further study is required to investigate the tracking of markers moving in 3-D in a treatment beam attenuated by moving patient anatomy. This will require a larger sensor with ROI read-out to maintain speed and a manageable data-rate.