BACKGROUND: i-STAT (i-STAT Corporation, Princeton NJ, USA), a hand-held point-of-care testing (POCT) analyzer with rapidity and minimal sample requirement, has the potential to bring about a significant impact on the management of neonates. However, there should be an overall deliberation of the routine use of i-STAT in the neonatal intensive care unit (NICU) as to whether it is technically reliable and cost-effective. The aim of this study was to assess the clinical aspects of the implementation of i-STAT in the NICU. METHODS: We surveyed physicians and nurses to measure the present status of POCT. We ana-lyzed 84 tests performed in the central laboratory, 88 tests by blood gas analyzer in NICU, and 95 tests by i-STAT for NICU patients. We investigated the indications, turnaround time (TAT), cycle time, and impact on patient care in each case during both pre- and post-i-STAT periods. Costs and user acceptability were also examined. RESULTS: Survey responders wanted rapid results but did not accept the responsibility for the quality of POCT. Turnaround time of i-STAT was shorter than that of the central laboratory, but did not make an impact on cycle time. The cost of i-STAT is 2.2 times higher than central laboratory cost, but the users were satisfied with i-STAT mainly because of its small sample volume and speed. Central laboratory testing volume decreased by 14.3% after the introduction of i-STAT. CONCLUSIONS: i-STAT may be acceptable in the NICU setting. However, the behavioral patterns of physicians need to be changed and a selective use of i-STAT is warranted to maximize its cost-effectiveness. Future studies on the clinical outcome are required to substantiate the potential role of i-STAT.