BackgroundPrevious intervention studies among night workers mainly focused on single interventions and found inconclusive evidence for effectiveness. A comprehensive intervention approach that includes individual and environmental components has been argued as important. Gaining insight into contributing factors for the implementation of interventions for night workers and effectiveness is important to distinguish between theory and programme failure. ObjectivesTo evaluate the effects and implementation of the PerfectFit@Night intervention to improve sleep, fatigue and recovery of night workers in healthcare, using the RE-AIM framework, which assesses reach, effectiveness, adoption, implementation and maintenance of interventions. DesignA prospective pre–post study design, with two measurements before and three and six months after the intervention. SettingTwelve different departments of a university hospital in the Netherlands. ParticipantsHealthcare workers working night shifts (n = 210). MethodsPerfectFit@Night consisted of environmental (provision of a powernap bed and healthy food, and workshop healthy rostering) and individual elements (e-learning and sleep coaching) and was implemented for three months in a phased manner. Questionnaires, logbooks and interview data were used. Effects of the intervention on sleep, fatigue and recovery were evaluated with mixed-effects models, and implementation factors of reach, adoption, implementation and maintenance were evaluated. ResultsNight shift-related insomnia (−11 %-points, 95 % CI: −19 %, −4 % at three months), need for recovery (β: −2.45, 95 % CI: −4.86, −0.03 at six months) and fatigue (OR: 0.46, 95 % CI: 0.25, 0.86 at six months) decreased significantly after the intervention. No changes were found for subjective sleep quality and sleep duration. Barriers and facilitators for implementation were identified for each intervention element at individual (e.g., dietary preferences), organisational (e.g., responsibilities at work) and workplace levels (e.g., location of power nap bed), and for the intervention itself (e.g., useful information in e-learning). Although satisfaction was high and continuation was preferred, embedding of the intervention in the daily routine was limited. Facilitators for future implementation include a positive attitude towards the intervention, clear guidelines regarding intervention elements, appointment of night workers as ambassadors, and suitable conditions in terms of work demands and for the intervention elements. ConclusionsThe multi-faceted PerfectFit@Night intervention reduced insomnia, fatigue and need for recovery in night workers in healthcare. The most important facilitators to improve the implementation of PerfectFit@Night exist at the organisational level (e.g., positive attitude within the culture and suitable work demands). Combining effect and implementation evaluation is crucial to identify barriers and facilitators that hamper or enhance intervention effects. Trial registrationThe study was registered in the Netherlands Trial Register on 17 January 2021 (trial number NL9224).