Sleep deprivation has been reported to affect intrinsic brain connectivity, notably reducing connectivity in the default mode network. Studies to date have however shown inconsistent effects, in many cases lacked monitoring of wakefulness, and largely included young participants. We investigated effects of sleep deprivation on intrinsic brain connectivity in young and older participants. Participants aged 20-30 (final n = 30) and 65-75 (final n = 23) years underwent partial sleep deprivation (3 h sleep) in a cross-over design, with two 8-minutes eyes-open resting state functional magnetic resonance imaging (fMRI) runs in each session, monitored by eye-tracking. We assessed intrinsic brain connectivity using independent components analysis (ICA) as well as seed-region analyses of functional connectivity, and also analysed global signal variability, regional homogeneity, and the amplitude of lowfrequency fluctuations. Sleep deprivation caused increased global signal variability. Changes in investigated resting state networks and in regional homogeneity were not statistically significant. Younger participants had higher connectivity in most examined networks, as well as higher regional homogeneity in areas including anterior and posterior cingulate cortex. In conclusion, we found that sleep deprivation caused increased global signal variability, and we speculate that this may be caused by wake-state instability.Sleep problems are a public health issue affecting about one third of the general population, of which about one in three reports serious sleep problems 1, 2 . Impaired or shortened sleep is a risk factor for mortality and for a number of diseases 3 , as well as accidents 4 . These risks appear to be mediated by impaired biological restoration/recovery 3 . Effects of sleep loss include an increasing number of short lapses of attention (microsleeps) 5 , as well as increased levels of EEG alpha and theta activity; slow, rolling eye movements; and increased subjective sleepiness 6 .Sleepiness is not well defined in terms of functional brain activation. Early studies using total sleep deprivation have shown reduced glucose uptake in large areas of the brain, including prefrontal and parietal areas 7 . Intrinsic connectivity, referring to functional connectivity in the resting state, i.e. when participants are not presented with any changing stimuli, has been investigated in several earlier studies. We have identified 21 earlier reports, based on 14 unique datasets . An in-depth review of this earlier literature is beyond the scope of this paper, and for this reason we have recently made available online an overview of these reports 29 . A consistent and robust finding is that sleep deprivation caused reduced connectivity within the default mode network and reduced anticorrelation to the task-positive network 8,10,22 . Other findings include increased regional homogeneity (ReHo) in different brain areas following sleep deprivation 9, 18 , changes in connectivity between the thalamus 12 and amygdala 13,14 and cor...