To investigate the effects of different body positions and head elevation angles on regional cerebral oxygen saturation in premature infants using near-infrared spectroscopy (NIRS).This was a prospective study of premature infants hospitalized. The position was changed each feeding cycle (2-3 h), in order: 15° prone, 15° supine, 0° prone, and 0° supine position. Regional cerebral oxygen saturation (rSO2) was measured after each position period using NIRS, over 7 days.Thirty-three premature infants were included in the analysis. Among them, 22 (66.7%) were male, and 11 (33.3%) were female. When placing the premature infants in the prone position, the regional cerebral oxygen saturation (rSO2) values were higher at head elevation 15° compared with 0° (main effect P < 0.001). When placing the premature infants in the supine position, rSO2 was higher at a head elevation of 15° compared with that at 0° (main effect P < 0.001). When placing the premature infants with a head elevation of 15°, rSO2 in the prone position was higher than that of the supine position (main effect P < 0.001). There was no difference between the prone and supine positions when placing the premature infants with the head elevation at 0° (P > 0.05).Higher cerebral rSO2 can be achieved when placing premature infants in the prone or supine position, with a 15° head elevation angle.Using a specific body position and head angle could improve the regional cerebral oxygen saturation of premature infants.