Background: Two understudied risk factors that have been linked to emotional difficulties in adolescence are chronotype and sleep deprivation. This study extended past research by using an experimental design to investigate the role of sleep deprivation and chronotype on emotion in adolescents. It was hypothesized that sleep deprivation and an evening chronotype would be associated with decreased positive affect (PA), increased negative affect (NA), and lower positivity ratios. Methods: Forty-seven healthy adolescents (aged 10-15 for girls, 11-16 for boys) participated in a sleep deprivation and a rested condition. A subsample of 24 adolescents was selected on the basis of extreme morningness or eveningness scores (based on outer quartiles of scores on the Children's Morningness-Eveningness Preferences Scale). PA and NA were measured using the Positive and Negative Affect Schedule for Children, and positivity ratios were calculated by dividing PA by NA. Results: Participants reported less positive affect and lower positivity ratios when sleep deprived, relative to when rested. Evening chronotypes reported less positive affect and lower positivity ratios than morning chronotypes in both rested and sleep deprivation conditions. Conclusions: These findings extend previous research by suggesting that adolescents are adversely impacted by sleep deprivation, and that an evening chronotype might serve as a useful marker of emotional vulnerability. Early intervention and prevention strategies can focus on improving sleep and on using chronotherapy principles to reduce eveningness.