Decoding paradoxical BOLD responses to transcranial ultrasound stimulation with concurrent optoacoustic magnetic resonance imaging

Focused transcranial ultrasound stimulation (TUS) can affect neural activity with high spatial precision, advancing noninvasive neuromodulation toward targeted treatment of brain disorders. Direct monitoring of TUS responses is crucial for ensuring optimal outcomes. Blood-oxygenation-level–dependent (BOLD) functional magnetic resonance imaging has primarily been used for studying TUS effects in both human and nonhuman primate brains. However, the physiology and mechanisms underlying BOLD remain largely unknown due to its highly convoluted nature. To address these limitations, we developed a hybrid system for concurrent optoacoustic and magnetic resonance imaging of TUS (OMRITUS) to comprehensively characterize the hemodynamic changes in murine brains. Our findings reveal paradoxical negative BOLD signals in the activated cortical regions, coupled with increased total hemoglobin levels simultaneously monitored with optoacoustic tomography. Multispectral optoacoustic readings further demonstrated a stronger increase in deoxygenated versus oxygenated hemoglobin, suggesting a potential molecular basis for the negative BOLD responses. OMRITUS enables the study of complex TUS-hemodynamic interactions, paving the way for precise neuromodulatory interventions.