Transfer functions linking neural calcium to single voxel functional ultrasound signal

Functional ultrasound imaging (fUS) is an emerging technique that detects changes of cerebral blood volume triggered by brain activation. Here, we investigate the extent to which fUS faithfully reports local neuronal activation by combining fUS and two-photon microscopy (2PM) in a co-registered single voxel brain volume. Using a machine-learning approach, we compute and validate transfer functions between dendritic calcium signals of specific neurons and vascular signals measured at both microscopic (2PM) and mesoscopic (fUS) levels. We find that transfer functions are robust across a wide range of stimulation paradigms and animals, and reveal a second vascular component of neurovascular coupling upon very strong stimulation. We propose that transfer functions can be considered as reliable quantitative reporters to follow neurovascular coupling dynamics. Neurovascular coupling refers to changes in cerebral blood flow in response to neuronal stimulation, but to what extent this change can report neuronal activation is not known. Here the authors develop transfer functions between neural calcium signals and functional ultrasound changes in blood volume in co-registered single voxel brain volumes.