Full-visibility 3D imaging of oxygenation and blood flow by simultaneous multispectral photoacoustic fluctuation imaging (MS-PAFI) and ultrasound Doppler.

We present a method and setup that provide complementary three-dimensional (3D) images of blood oxygenation (via quantitative photoacoustic imaging) and blood flow dynamics (via ultrasound Doppler). The proposed approach is label-free and exploits blood-induced fluctuations, and is implemented on a sparse array with only 256 elements, driven with a commercially available ultrasound electronics. We first implement 3D photoacoustic fluctuation imaging (PAFI) to image chicken embryo, and obtain full-visibility images of the vascular morphology. We obtain simultaneously 3D ultrasound power Doppler with a comparable image quality. We then introduce multispectral photoacoustic fluctuation imaging (MS-PAFI), and demonstrate that it can provide quantitative measurements of the absorbed optical energy density with full visibility and enhanced contrast, as compared to conventional delay-and-sum multispectral photoacoustic imaging. We finally showcase the synergy and complementarity between MS-PAFI, which provides 3D quantitative oxygenation (SO[Formula: see text]) imaging, and 3D ultrasound Doppler, which provides quantitative information on blood flow dynamics. MS-PAFI represents a promising alternative to model-based inversions with the advantage of resolving all the visibility artefacts without prior and regularization, by use of a straightforward processing scheme.