AbstractBackgroundNursing checklists have been shown to improve communication, reduce the occurrence of adverse events, and promote safe, quality care in different care settings. However, to date, there is no validated patient care safety checklist for nurses caring for infants in Neonatal Intensive Care Units (NICU).AimTo describe development and content validation of the “Safe Nursing Care Checklist for Infants Hospitalized in the Neonatal Intensive Care Unit”.Study DesignOnline Survey.MethodsBased upon an integrative literature review, we developed a checklist focused on safe nursing care for infants in the NICU. Nursing experts participated in three rounds of a content validation process where they rated the items online. An agreement level ≥0.90 was required for inclusion in the checklist. Forty‐ three expert nurses with experience working in the NICU and who were certified in neonatal nursing or had a master's or doctoratal degree in child health provided content validation of the patient care checklist.ResultsThe final checklist contained 45 items with content validation index scores greater than 90%. The instrument was structured into six dimensions including patient identification, effective communication, medication safety, infection prevention, fall prevention, and pressure injuries/skin injuries prevention.ConclusionContent validity was established for the “Safe Nursing Care Checklist for Infants Hospitalized in the Neonatal Intensive Care Unit” which can identify strengths and weaknesses in safe nursing care for infants in the NICU as well as direct educational interventions to promote nursing care based on scientific evidence.Relevance to Clinical PracticeThis checklist can potentially be used by bedside nurses to promote provision of safe care to infants in the NICU and to guide corrective strategies and encourage evidence‐based decision‐making. Validation in the clinical setting is needed.