Volumetric Functional Ultrasound Imaging in Macaques

Linking circuit level activity to large scale functional organization requires imaging methods combining high spatial resolution, broad coverage, and single trial sensitivity. We present volumetric functional ultrasound imaging (3D-fUS) in behaving macaques, enabling imaging of ~1 cm³ cortical volumes at high spatiotemporal resolution (100 × 150 × 150 μm³ voxels, 1.67 Hz). Visually evoked responses were reliably detected at the level of single trials and single voxels, substantially reducing experimental time. To enable model-based analyses analogous to functional magnetic resonance imaging (fMRI), we estimated a fUS hemodynamic response function (fUS-HRF) that was consistent across subjects, cortical areas, and visual stimuli and was well approximated by a gamma function. Compared with fMRI HRFs, the fUS-HRF exhibited faster dynamics, enabling shorter and more closely spaced stimulus presentations. Together, these results establish 3D-fUS as a fast, volumetric, and circuit relevant imaging modality for efficient investigation of distributed cortical dynamics in primates.