Background: Due to the progress in neonatology, in particular, in the past three decades, the mortality rate among patients of intensive care units has decreased. However, this is connected not only with newborns needing to stay longer in the unit, but also with the exposure of newborns to many painful procedures and stresses. Lack of or insufficient pain prevention has a negative impact on the sensory or locomotor development of newborns. Despite the presence of guidelines based on scientific evidence, the use of pharmacological and non-pharmacological pain-management methods in newborns is still insufficient. Aim: The aim of the study was to: identify the knowledge nurses/midwives have of recommended non-pharmacological and/or pharmacological methods, in particular, in relation to medical intervention procedures; assess the interventions for pain relief applied by midwives/nurses most often in their clinical practice; examine the role of age, general work experience, education level and years of work of medical professionals on a neonatal ward, as well as the referral level of a unit, versus the application of pharmacological and non-pharmacological methods. Methods: A descriptive and quantitative study conducted in 2019 among Polish nurses/midwives working at neonatal intensive care units. Results: The analysis of the material reflected the deficit of knowledge and the insufficient daily use of recommended pain-relief measures among the respondents. Conclusions: The interpretation of data indicates that despite the clear and easily available recommendations of scientific societies concerning the mode of conduct in particular medical procedures, medical personnel do not apply those recommendations in their everyday practice. It is necessary to plan and implement education strategies for nurses/midwives on standard pain-management interventions during painful medical procedures.