Spatial Relationship between Flavoprotein Fluorescence and the Hemodynamic Response in the Primary Visual Cortex of Alert Macaque Monkeys

Flavoprotein fluorescence imaging (FFI) is a novel intrinsic optical signal that is steadily gaining ground as a valuable imaging tool in neuroscience research due to its closer relationship with local metabolism relative to the more commonly used hemodynamic signals. We have developed a technique for FFI imaging in the primary visual cortex (V1) of alert monkeys. Due to the nature of neurovascular coupling, hemodynamic signals are known to spread beyond the locus of metabolic activity. To determine whether FFI signals could provide a more focal measure of cortical activity in alert animals, we compared FFI and hemodynamic point spreads (i.e. responses to a minimal visual stimulus) and functional mapping signals over V1 in macaques performing simple fixation tasks. FFI responses were biphasic, with an early and focal fluorescence increase followed by a delayed and spatially broader fluorescence decrease. As expected, the early fluorescence increase, indicating increased local oxidative metabolism, was somewhat narrower than the simultaneously observed hemodynamic response. However, the later FFI decrease was broader than the hemodynamic response and started prior to the cessation of visual stimulation suggesting different mechanisms underlying the two phases of the fluorescence signal. FFI mapping signals were free of vascular artifacts and comparable in amplitude to hemodynamic mapping signals. These results indicate that the FFI response may be a more local and direct indicator of cortical metabolism than the hemodynamic response in alert animals.