Abstract: Objective To explore the effects of modified localization method on the radial artery puncture in neonatal intensive care unit. Methods A total of 232 neonates who were admitted into neonatal intensive care unit from January 2017 to November 2018 for arterial blood collection were selected. They were randomly divided into two groups (n=116): a research group and a control group. A modified localization method was used in the research group for radial artery puncture, while the traditional localization method was used in the control group. The successful rate of first puncture, the successful and failure rates of multiple puncture, the time required for successful puncture and the incidence of complications after puncture were monitored. Results Compared the control group, the research group presented remarkable increases in the successful rate of first puncture, and decreases in the time required for successful puncture, the incidence of complications and failure rate of puncture (P<0.05). Conclusions The modified localization method has a high successful rate of puncture and requires short time, with few complications for radial artery puncture in neonatal intensive care unit, which is of importance for clinical nursing care.