In the NICU, systematic exposure to sound-pressure above the recommended level can affect both neonates and staff. This study aimed to evaluate the sound pressure levels in three Portuguese NICUs and the noise perceptions of staff. The measurements were performed with a sound-level meter, considering the location of the main sources of noise and the layout of the units. A questionnaire was applied to assess noise perceptions of professionals. Among the staff, 41.1% classified the environment (regarding noise) as "slightly uncomfortable"; 48.4% considered it as "acceptable." The majority (55.5%) considered "equipment" the most annoying source of noise. The results showed that noise levels were excessive in all the evaluated areas of the NICUs, exceeding international guidelines, with levels ranging between 48.7 dBA to 71.7 dBA. Overall, there is a need for more research to verify the effectiveness of some actions and strategies to reduce the effect of noise in the NICU.