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. 2020 Apr 7;10:6007. doi: 10.1038/s41598-020-62898-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Chicken embryo CAM imaging with microscopic imaging validation. (a) An ex ovo chicken embryo imaged with a high-frequency transducer through an acoustic window in the side of container. (b) Co-registered optical imaging of the ultrasound field-of-view. (c) Ultrasound MB data was used to generate ULM super-resolution images. (d) Conventional ULM image reconstruction without microbubble separation. Lines 1 and 2 denote two manually selected vessel cross-sections of interest. (e) ULM intensity image with microbubble separation method. (f) ULM velocity map generated with microbubble separation. The arrows highlight adjacent vessels with opposing flow directions. (g) Magnified section of CAM with arrows pointing out microvasculature that was not detected by conventional ULM. (h) Intensity and velocity profiles of CAM vessels from manually selected line segments.