Introduction: Sleep deprivation is a common problem that may have serious consequences. In this study, functional magnetic resonance imaging (fMRI), a technique frequently used to study networks in the brain, was used to investigate the resting state of the sleep deprived brain, in order to discover whether this state affects the intrinsic connectivity and the global signal variability (GSV). Aims: To investigate whether GSV increases with sleep deprivation. Material and Methods: In this cross over study 18 healthy participants, age 20 – 30, underwent in randomized order resting-state fMRI for 20min before and after 24h sleep deprivation. We extracted the global signal, calculated the standard deviation per participant, and analysed it with respect to sleep depraved yes/no, head motion, eyes open/closed and self-evaluation of sleepiness using Karolinska Sleepiness Score (KSS). Results: We found that GSV was higher during sleep deprivation (0.3362 ± 0.0241, p<0.0001) without KSS data. With KSS, sleep deprivation was not significant (0.0619 ± 0.1145, p=0.5889). High KSS rating had a significant effect on GSV (0.1497 ± 0.0409, p=0.0003), as had head motion (1.7974 ± 0.1539, p<0.0001). There was no significant difference between having eyes open or closed (0.0126 ± 0.0578, p=0.8278), and no significant increase for each time period of 20s in the scanner (0.0065 ± 0.0021, p=0.0029). Conclusions: We found that the global signal variation is increased by sleep deprivation and sleepiness. More specific conclusions cannot be made from our data so far.