Aims: The transition to parenthood is a challenging learning process for parents. The process can become more stressful, especially for mothers when the newborn baby is admitted to the intensive care unit for some reason. Methods: This study was conducted to determine the relationship between participation in caregiving and maternal stress and maternal role among mothers with babies in the neonatal intensive care unit in Turkey. Data were collected after the mothers had been providing care in the NICU for at least 24 h, with the babies being on average 5.02 ± 9.96 days old at the time of data collection. Data collection tools included the Assessment of Mothers’ Participation in Caregiving Scale to evaluate the level of caregiving involvement, the Semantic Differential Scale–Myself as a Mother to assess maternal role perception, and the NICU Parental Stressor Scale to measure stress levels related to the NICU environment. Results: A total of 193 women whose babies were hospitalized in the neonatal intensive care unit were included in the study. The mean scores of the participants were 7.04 ± 0.64 on the Assessment of Mothers' Participation in Caregiving Scale, 43.33 ± 6.40 on the Semantic Differential Scale, and 3.29 ± 1.07 on the NICU Parental Stressor Scale. It was determined that there was a negative significant relationship between participation in caregiving and the mean scores on the total and sight and sounds and relationship with the infant and maternal role sub-dimensions of the Parental Stressor Scale (p < 0.05). Conclusion: Considering the intensive care conditions, mothers should be allowed to see their babies as much as possible, and they should be supported to participate in the care.