All perfusion-based imaging modalities depend on the relationship between neuronal and vascular activity. However, the relationship between stimulus and response was never fully characterized. With the use of optical imaging (intrinsic signals and intravascular fluorescent dyes) during repetitive stimulation paradigms, we observed reduced responses with temporally close stimuli. Cortical evoked potentials, however, did not produce the same reduced responsiveness. We therefore termed these intervals of reduced responsiveness "refractory periods." During these refractory periods an ability to respond was retained, but at a near 60% reduction in the initial magnitude. Although increasing the initial stimulus duration lengthened the observed refractory periods, significantly novel or temporally spaced stimuli overcame them. We observed this phenomenon in both rodent and human subjects in somatosensory and auditory cortices. These results have significant implications for understanding the capacities, mechanisms, and distributions of neurovascular coupling and thereby possess relevance to all perfusion-dependent functional imaging techniques.