ABSTRACT It is known that working in the shift system, especially the night shift, affects physical, mental, and social well-being. We investigated the changes in the inner retinal layers and choroidal layer of the eyes of nurses working night and day shifts using optical coherence tomography (OCT). We also explored the effect of night shift work on metacognition and the relationships between these variables. A total of 79 nurses participated in the study, of whom 40 worked night shifts. The researcher gave the participants sociodemographic information and the Metacognition Questionnaire-30 (MCQ-30) form. Retinal nerve fiber layer (RNFL) thickness, ganglion cell layer (GCL) thickness, inner nuclear layer (INL) thickness, inner plexiform layer (IPL) thickness, central macular thickness (CMT), and subfoveal choroidal thickness (SFCT) were measured with OCT. It was found that the level of metacognitive activity associated with cognitive confidence was higher (p = 0.044) for nurses who worked night shifts and that the level of metacognitive activity associated with cognitive awareness was lower (p = 0.015) for nurses who worked night shifts. RNFL-nasal superior (NS) thickness was lower in night shift workers than the day shift group (p = 0.017). Our study revealed significant relationships between metacognition and the OCT findings among night and day shift workers. Our study revealed that RNFL measurements and metacognitive activity may differ and there may be a relationship between these parameters in nurses who work shifts. Further research is needed to investigate the long-term effects of night shift work on retinal health.