The aim of this study was principally to assess the impact of sleep deprivation on interference performance in short Stroop tasks (Color-Word, Emotional, and Specific) and on subjective anxiety. Subjective sleepiness and performance on a psychomotor sustained attention task were also investigated to validate our protocol of sleep deprivation. Twelve healthy young subjects were tested at four-hourly intervals through a 36-h period of wakefulness under a constant routine protocol. Analyses of variance for repeated measurements revealed that self-assessment of sleepiness on a visual analogue scale as well as mean reaction time performance on the sustained attention task, both for the first minute and for 10 min of testing, were worsened by sleep deprivation. Analyses revealed an increase in self-reported anxiety scores on the STAI questionnaire but did not reveal any significant effect after sleep deprivation either on indexes of interference or on accuracy in Stroop tasks. However, analyses showed sensitivity to circadian effect on verbal reaction times in the threat-related (Emotional) and sleep-related (Specific) Stroop tasks. We concluded that 36 h of prolonged wakefulness affect self-reported anxiety and Emotional Stroop task resulting in a cognitive slowing. Moreover, total sleep deprivation does not affect interference control in any of the three short Stroop tasks.