Removal of the interplane septa and configuration of a typical multislice PET scanner to accept all possible coincidence lines of response leads to a fivefold increase in sensitivity. This can be of value in regional CBF studies using bolus 15O-labeled water injections, allowing the injected dose to be reduced by a factor of 4, while maintaining the same number of noise equivalent counts. Thus, for a given cumulative dose limit, four times as many studies can be performed in a single subject. Data from the three-dimensional Hoffman brain phantom, closely matched to count rates seen in human studies, show that for an identical cumulative dose, the noise in subtraction (stimulus minus baseline) images can be reduced by a factor of 2 using three-dimensional data acquisition, with appropriate fractionation of the dose. This improvement is dependent on axial position due to the sensitivity characteristics of three-dimensional scans; however, there is a significant gain in the signal-to-noise ratio (S/N) in all image planes. Studies performed in a human subject demonstrate how the factor of 2 gain in S/N leads to improved detectability of activation sites in PET subtraction images.