Background: The role of neonatal pain on the developing nervous system is not completely understood, but evidence suggests that sensory pathways are influenced by an infants pain experience. Research has shown that an infants previous pain experiences lead to an increased, and likely abnormal, response to subsequent painful stimuli. We are working to improve neonatal pain detection through automated devices that continuously monitor an infant. The current study outlines some of the initial steps we have taken to evaluate Near Infrared Spectroscopy (NIRS) as a technology to detect neonatal pain. Our findings may provide neonatal intensive care unit (NICU) practitioners with the data necessary to monitor and perhaps better manage an abnormal pain response. Methods: A prospective pilot study was conducted to evaluate nociceptive evoked cortical activity in preterm infants. NIRS data were recorded for approximately 10 minutes prior to an acute painful procedure and for approximately 10 minutes after the procedure. Individual data collection events were performed at a weekly maximum frequency. Eligible infants included those admitted to the Tampa General Hospital (TGH) NICU with a birth gestational age of less than 37 weeks. Results: A total of 15 infants were enrolled and 25 individual studies were completed. Analysis demonstrated a statistically significant difference between the median of the pre- and post-painful procedure data sets in each infants first NIRS collection (p value = 0.01). Conclusions: Initial analysis shows NIRS may be useful in detecting acute pain. An acute painful procedure is typically followed by a negative deflection in NIRS readings.