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Background: Infection prevention (IP) behaviors such as hand hygiene (HH) and mobile device disinfection are important to reduce the risk of infection transmission from both family members and hospital staff to critically ill neonates. Purpose: To inform the design of educational interventions to improve both patient family and staff IP behaviors, we engaged separate groups of nurses and family members to understand perceptions about the spread of infection and barriers to implementing effective IP strategies. Methods: This was a qualitative study using focus groups to gather data from neonatal nurses and patient family members. Data were triangulated with hospital-wide survey data and analyzed using inductive content analysis. Results: Twelve nurses and 4 patient family members participated. Themes related to communication about IP between staff and family members emerged: stakeholders expressed discomfort with the timing and nature of just-in-time HH education. These communication challenges contributed to stress levels within the neonatal intensive care unit. This finding was reflected in the hospital-wide survey. Implications for Practice and Research: Steps should be taken to improve communication about IP behaviors between patient family members and frontline staff. Reducing nurse burden of providing just-in-time HH reminders to patient family members through increased IP education may decrease stress and facilitate IP behaviors. This has the potential to decrease infection spread and improve patient outcomes. The development of interventions targeting stakeholder communication is therefore warranted, but additional research is needed to understand the timing and process for delivery of the educational material. Video Abstract available for more insights from the authors (see the Video, Supplemental Digital Content available at http://www.w3.org/1999/xlink). [ABSTRACT FROM AUTHOR]