The study explores the impact of light conditions on driver sleepiness. In a driving simulator experiment, 20 drivers drove both during daytime in an alert condition and then later at night after being awake since early morning. Light conditions were manipulated by driving in a simulated nighttime scenario in a dark room (1 lx) versus driving in simulated daylight in a room lit with light emitting diodes combining blue light with a yellow phosphor giving a two peaked spectrum (212 lx). Both the daylight and the darkness scenarios were driven daytime and nighttime in a 2 × 2 design. Sleepiness was measured during the four 1-hour drives in terms of subjective sleepiness ratings, divided attention ability, driving performance, heart rate variability and blink behaviour. Significant differences were found in all measured sleepiness indicators between the daytime and nighttime drives, and in most indicators for time-on- task. No significant main effects were found between simulated daylight and darkness. A psychomotor vigilance test conducted before and after each drive also showed no significant effects for lighting condition. Further research, preferably using longitudinal studies in more realistic settings on real roads, is needed to determine which behaviours and which cognitive processes that are affected when driving in daylight versus darkness.