Three-dimensional visualization and improved quantification with super-resolution ultrasound imaging - validation framework for analysis of microvascular morphology using a chicken embryo model.

The purpose of this study was to improve the morphological analysis of microvascular networks depicted in three-dimensional (3D) super-resolution ultrasound (SR-US) images. This was supported by qualitative and quantitative validation by comparison to matched brightfield microscopy and traditional B-mode ultrasound (US) images. Contrast-enhanced US (CEUS) images were collected using a preclinical US scanner (Vevo 3100, FUJIFILM VisualSonics Inc.) equipped with an MX250 linear array transducer. CEUS imaging was performed after administration of a microbubble (MB) contrast agent into the vitelline network of a developing chicken embryo. Volume data was collected by mechanically scanning the US transducer throughout a tissue volume-of-interest in 90μm step increments. CEUS images were collected at each increment and stored as in-phase/quadrature data (2000 frames at 152 frames per sec). SR-US images were created for each cross-sectional plane using established data processing methods. All SR-US images were then used to reconstruct a final 3D volume for vessel diameter (VD) quantification and for surface rendering. VD quantification from the 3D SR-US data exhibited an average error of 6.1% ± 6.0% when compared with matched brightfield microscopy images, whereas measurements from B-mode US images had an average error of 77.1% ± 68.9%. Volume and surface renderings in 3D space enabled qualitative validation and improved visualization of small vessels below the axial resolution of the US system. Overall, 3D SR-US image reconstructions depicted the microvascular network of the developing chicken embryos. Improved visualization of isolated vessels and quantification of microvascular morphology from SR-US images achieved a considerably greater accuracy compared to B-mode US measurements.