The spatial pattern of activation in response to multiple whisker stimulation was studied using high-resolution laser Doppler (LD) imaging in urethane-anesthetized rats. LD flux change representing cerebral blood flow (CBF) responses were analyzed from a single trial or after averaging a number of similar trials. CBF change in a single trial was observed predominantly over pixels having low baseline flux values (microvessels), which included diffuse circular patterns of activation 400-800 microm in diameter similar to the histological dimensions of individual barrels established in the layer IV of the rat somatosensory cortex. The overall activation pattern varied considerably between each trial (only about 9-10% overlap); however, the diffuse circular pattern of activation was reproducible in every single trial within and across all rats. With trial averaging, no significant increase was observed in the outer boundary of activation, but the number of activated pixels increased within the diffuse circular patterns of activation. Emergence of further active pixels primarily within the diffuse circular regions of activity with trial averaging indicates distinct CBF responses in the septal and barrel regions, with a lesser LD signal to noise ratio in the barrel core.