Acoustically probing biological tissues with light or sound, photoacoustic and ultrasound imaging can provide anatomical, functional, and/or molecular information at depths far beyond the optical diffusion limit. However, most photoacoustic and ultrasound imaging systems rely on linear-array transducers with elevational focusing and are limited to two-dimensional imaging with anisotropic resolutions. Here, we present three-dimensional diffractive acoustic tomography (3D-DAT), which uses an off-the-shelf linear-array transducer with single-slit acoustic diffraction. Without jeopardizing its accessibility by general users, 3D-DAT has achieved simultaneous 3D photoacoustic and ultrasound imaging with optimal imaging performance in deep tissues, providing near-isotropic resolutions, high imaging speed, and a large field-of-view, as well as enhanced quantitative accuracy and detection sensitivity. Moreover, powered by the fast focal line volumetric reconstruction, 3D-DAT has achieved 50-fold faster reconstruction times than traditional photoacoustic imaging reconstruction. Using 3D-DAT on small animal models, we mapped the distribution of the biliverdin-binding serpin complex in glassfrogs, tracked gold nanoparticle accumulation in a mouse tumor model, imaged genetically-encoded photoswitchable tumors, and investigated polyfluoroalkyl substances exposure on developing embryos. With its enhanced imaging performance and high accessibility, 3D-DAT may find broad applications in fundamental life sciences and biomedical research. The authors demonstrate integration of a single-slit diffraction method into linear-array transducers for advancing 3D photoacoustic and ultrasound imaging in biological tissues. The method enhances spatial resolution and signal-to-noise ratio while maintaining low-cost and practicality.
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