Purpose: The authors have recently developed a cine-mode T2*/T1-weighted 4D-MRI technique and a sequential-mode T2-weighted 4D-MRI technique for imaging respiratory motion. This study aims at investigating which 4D-MRI image acquisition mode, cine or sequential, provides more accurate measurement of organ motion during respiration. Methods: A 4D digital extended cardiac-torso (XCAT) human phantom with a hypothesized tumor was used to simulate the image acquisition and the 4D-MRI reconstruction. The respiratory motion was controlled by the given breathing signal profiles. The tumor was manipulated to move continuously with the surrounding tissue. The motion trajectories were measured from both sequential-and cine-mode 4D-MRI images. The measured trajectories were compared with the average trajectory calculated from the input profiles, which was used as references. The error in 4D-MRI tumor motion trajectory (E) was determined. In addition, the corresponding respiratory motion amplitudes of all the selected 2D images for 4D reconstruction were recorded. Each of the amplitude was compared with the amplitude of its associated bin on the average breathing curve. The mean differences from the average breathing curve across all slice positions (D) were calculated. A total of 500 simulated respiratory profiles with a wide range of irregularity (Ir) were used to investigate the relationship between D and Ir. Furthermore, statistical analysis of E and D using XCAT controlled by 20 cancer patients' breathing profiles was conducted. Wilcoxon Signed Rank test was conducted to compare two modes. Results: D increased faster for cine-mode (D = 1.17 × Ir + 0.23) than sequential-mode (D = 0.47 × Ir + 0.23) as irregularity increased. For the XCAT study using 20 cancer patients' breathing profiles, the median E values were significantly different: 0.12 and 0.10 cm for cine-and sequential-modes, respectively, with a p-value of 0.02. The median D values were significantly different: 0.47 and 0.24 cm for cine-and sequential-modes, respectively, with a p-value < 0.001. Conclusions: Respiratory motion measurement may be more accurate and less susceptible to breathing irregularity in sequential-mode 4D-MRI than that in cine-mode 4D-MRI. C 2016 American Association of Physicists in Medicine. [http://dx