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The technique allows new insights into microvascular anatomy and physiology and the development of disease-related microvascular abnormalities. However, the microvascular anatomy is intricate and challenging to depict with the currently available imaging techniques, and validation of the microvascular structures of deeper-lying organs obtained with SRUS remains difficult. Our study aimed to directly compare the vascular anatomy in two in vivo 2D SRUS images of a Sprague\u2013Dawley rat kidney with ex vivo <jats:italic>\u03bc<\/jats:italic>CT of the same kidney. Co-registering the SRUS images to the <jats:italic>\u03bc<\/jats:italic>CT volume revealed visually very similar vascular features of vessels ranging from\u2009~\u2009100 to 1300\u00a0\u03bcm in diameter and illustrated a high level of vessel branching complexity captured in the 2D SRUS images. Additionally, it was shown that it is difficult to use <jats:italic>\u03bc<\/jats:italic>CT data of a whole rat kidney specimen to validate the super-resolution capability of our ultrasound scans, i.e<jats:italic>.<\/jats:italic>, validating the actual microvasculature of the rat kidney. Lastly, by comparing the two imaging modalities, fundamental challenges for 2D SRUS were demonstrated, including the complexity of projecting a 3D vessel network into 2D. These challenges should be considered when interpreting clinical or preclinical SRUS data in future studies.<\/jats:p>","DOI":"10.1038\/s41598-021-03726-6","type":"journal-article","created":{"date-parts":[[2021,12,21]],"date-time":"2021-12-21T11:03:53Z","timestamp":1640084633000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["Evaluation of 2D super-resolution ultrasound imaging of the rat renal vasculature using ex vivo micro-computed tomography"],"prefix":"10.1038","volume":"11","author":[{"given":"Sofie Bech","family":"Andersen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iman","family":"Taghavi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hans Martin","family":"Kjer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stinne Byrholdt","family":"S\u00f8gaard","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carsten","family":"Gundlach","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vedrana Andersen","family":"Dahl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael Bachmann","family":"Nielsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anders Bjorholm","family":"Dahl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00f8rgen Arendt","family":"Jensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Charlotte Mehlin","family":"S\u00f8rensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,12,21]]},"reference":[{"key":"3726_CR1","doi-asserted-by":"crossref","first-page":"499","DOI":"10.1038\/nature16066","volume":"527","author":"C Errico","year":"2015","unstructured":"Errico, C. et al. 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