STUDY OBJECTIVES:Childhood sleep disorders are consistently shown to affect behavior and cognition, but first-night effects on these measures are generally unknown. We sought to examine how sleep in the home versus the laboratory differed among healthy toddlers and how such differences relate to standardized scores on assessments the morning following polysomnography. DESIGN, SETTING, AND PARTICIPANTS:Twenty healthy 14-month-olds wore actigraphs during nighttime sleep at home for 5 nights preceding and during standard overnight laboratory polysomnography. The Bayley Scales of Infant Development (BSID-II) were administered once the morning after polysomnography. MEASUREMENTS AND RESULTS:All subjects had normal polysomnography. Sleep-start times at home and during polysomnography did not differ, whereas, during polysomnography, subjects awoke earlier (p = .008, d = .58), their total sleep time (p <. 001, d = 1.1) and sleep efficiency (p = .004, d = .57) were reduced, and they had shorter sleep-bout lengths (p = .004, d = .03), less immobility (p = .003, d = .62), and greater average activity during sleep (p <. 001, d = .98). Standardized assessments were not affected by differences between home and polysomnography night sleep, but children with greater emotional regulation difficulty had a lower percentage of immobility (r = -0.67, p = .001) and increased sleep fragmentation (r = -0.60, p = .005) during polysomnography. CONCLUSIONS:Although sleep-onset times were preserved, sleep in the laboratory was disrupted, compared with at home. These differences did not affect standardized scores, but the magnitude of the difference was associated with worse emotional regulation. The effects of sleep disturbance during polysomnography, or the influence of poor emotional regulation on sleep in the laboratory, should be considered in studies of young children.