Versatile Vasculature Chips for Ultrasound Localization Microscopy

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Abstract Ultrasound localization microscopy (ULM) has revolutionized microvasculature imaging by surpassing the diffraction limit via microbubbles. ULM demonstrates exceptional potential to resolve micrometer-scale vascular structures in both preclinical and clinical studies. However, its performance evaluation remains challenging primarily due to the lack of reference microvascular phantoms featuring micrometer-scale, hierarchical branches, and realistic vascular structures. Inspired by microfluidic chip techniques, we present an organ-on-a-chip protocol for fabricating agarose-based micro-vessel network phantoms with ground truth. The vasculature pattern offers design versatility, enabling on-demand customization. We experimentally demonstrated the feasibility of the vasculature phantom using two adapted patterns. The first was a leaf pattern, which exhibited intrinsic quasi-two-dimensional venation network with hierarchical and branching channels similar to animal vasculature. The second was a kidney pattern, which was based on a two-dimensional projection of real human vasculature obtained from micro computed tomography. The microbubble solution was perfused into the phantoms by capillary force and gravity. The ULM-reconstructed vasculature maps agreed well with the ground truth. ULM achieved a high sensitivity of 0.97 and 0.95, but a low precision of 0.37 and 0.60, for the leaf and kidney phantom, respectively. The results indicated the capability of ULM to reconstruct vessel structures while making many false positive predictions. The proposed protocol holds significant promise for the development and optimization of ultrasound microvascular imaging techniques.

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