Background: Multispectral optical imaging has the capability of resoling hemoglobin, lipid,and water. Volumetric multispectral optoacoustic tomography (MSOT) is a hybrid imaging technique that provides a unique combination of functional and molecular contrast with real-time handheld imaging.Purpose: To investigate whether volumetric MSOT can provide real-time assessment of the anatomic and functional status of the human carotid artery bifurcation nonoinvasively.Materials and Methods: Imaging of healthy volunteers (n = 16) was performed with a custom-designed handheld volumetric MSOT scanner capable. of high-spatial-resolution (approximately 200 mu m) and real-time (10 volumes/sec) three-dimensional imaging, while further providing spectrosccopic capacity through fast tuning of the excitation light wavelength. For comparison and anatomic cross-validation, volunteers were also scanned with clinical B-mode US.Results: Volumetric MSOT achieved real-time imaging and characterization of the entire carotid bifurcation area across three dimensions simultaneously captured in a single volumetric image frame. Analysis of the acquired data further showed that a higher contrast-to-noise ratio can be achieved for wavelengths corresponding to a high optical absorption of oxygenated hemoglobin.Conclusion: The human carotid artery was visualized by using handheld volumetric multispectral optoacoustic tomography. This imaging approach is less prone to motion artifacts than are the conventional clinical imaging methods, holding promise for providing additional image-based biomarkers for noninvasive label-free assesment of carotid artery disease. (C) RSNA, 2019