We investigated temporal MR signal changes in the peripheral lung and proximal pulmonary vessels during the entire cardiac cycle in order to evaluate the characteristics of the diastolic-systolic subtraction method in the lung. In eight healthy volunteers free of lung diseases, changes in the MR signal during one breath-hold were investigated with the multiple ECG-triggered half-Fourier single-shot fast-spin echo (SS-FSE) technique. The signal intensity-time course curve in the lung showed that biphasic signals decreased 20% to 47% at systole and 5% to 33% at mid-diastole, measured against the maximum signals at late diastole. This signal decrease in the peripheral lung was correlated to that in the proximal pulmonary vessels during an entire cardiac cycle (r=0.667 to 1.000). The best visualization of the lung was obtained at late diastole, when the intra-vascular flow in the lung was expected to be stagnant. Compared with the late diastolic SS-FSE images, the late diastolic-systolic subtracted SS-FSE images improved the signal-to-noise ratio in the lung as well as the signal-intensity ratio of the peripheral lung to surrounding tissues. Although the flow-induced signal dephasing in the lung was completely unavoidable and its amount was unpredictable even at late diastole, the diastolic-systolic subtracted SS-FSE images showed the relative differences in flow alteration during the cardiac cycle between the images at diastole and those at systole. The main characteristic of diastolic-systolic subtracted SS-FSE was the enhancement of visibility of cardiac-dependent signal changes in the lung due to the alteration in pulsatile flow.