Abstract: Background: In 1989 the World Health Organization and UNICEF introduced the “Ten Steps” for successful breastfeeding. One step suggests that a mother and her newborn baby should remain together day and night during the hospital stay. The purpose of this study was to investigate, first, whether or not mothers in our hospital roomed-in with their babies at night, second, the attitudes of mothers toward night rooming-in and their feelings of closeness to their babies, and third, how mothers perceived hospital staff attitudes toward night rooming-in. Methods: All mothers ( n = 132) of Nordic ancestry and with good knowledge of the Swedish language, who were admitted to the maternity wards during a 2-week period at Karolinska University Hospital, Stockholm, Sweden, answered a questionnaire on demographic background data and their current night rooming-in practices, including an attitude scale. Results: Most study mothers were positive toward night rooming-in, regardless of whether they had roomed in with their babies at night (93% positive) or not (73% positive). Mothers who had not roomed-in with their babies were more likely to perceive that the staff believed their babies should stay in the nursery compared with those mothers who practiced night rooming-in (z = −2.733, p = 0.006). Mothers not rooming-in with their babies scored closeness to their babies as less important than those mothers who roomed-in with their babies (z = −3.780, p = 0.0002); they also were more worried about their own and their babies’ sleep (z = −2.321, p = 0.02) and disturbing noises (z = −3.487, p = 0.0005). Conclusions: Mothers who left their babies in the nursery at night more often perceived that the staff believed their babies should stay in the nursery, rating closeness between mother and infant lower. Hence, negative staff attitudes toward night rooming-in may implicitly suggest to mothers that closeness between mothers and babies is not important.