Neuronal activity elicits metabolic and vascular responses, during which oxygen is first consumed and then supplied to the tissue via an increase in cerebral blood flow. Understanding the spatial and temporal dynamics of blood and tissue oxygen (Symbol) responses following neuronal activity is crucial for understanding the physiological basis of functional neuroimaging signals. However, our knowledge is limited because previousSymbolmeasurements have been made at low temporal resolution (>100 ms). Here we recordedSymbolat high temporal resolution (1 ms), simultaneously with co-localized field potentials, at several cortical depths from the whisker region of the somatosensory cortex in anaesthetized rats and mice. Stimulation of the whiskers produced rapid, laminar-specific changes inSymbol. PositiveSymbolresponses (i.e. increases) were observed in the superficial layers within 50 ms of stimulus onset, faster than previously reported. NegativeSymbolresponses (i.e. decreases) were observed in the deeper layers, with maximal amplitude in layer IV, within 40 ms of stimulus onset. The amplitude of the negative, but not the positive,Symbolresponse correlated with local field potential amplitude. Disruption of neurovascular coupling, via nitric oxide synthase inhibition, abolished positiveSymbolresponses to whisker stimulation in the superficial layers and increased negativeSymbolresponses in all layers. Our data show thatSymbolresponses occur rapidly following neuronal activity and are laminar dependent.