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. 2022 Jan 12;12:619. doi: 10.1038/s41598-021-04712-8

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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ULM image clustering and distributional analysis. (A) Super-resolution vessel maps were clustered based on microbubble accumulation density using K-means clustering with 6 clusters. (B) Splitting the clustered map into separate high-valued and low-valued images allowed for the generation of large vessel and small vessel representations of the ULM data, respectively. (C) The mean intervessel distance across the entire coronel hemisphere for the large vessel map could be calculated, along with the microvascular density for the small vessel map, for all mice in each age group. No significant difference was found between the age groups for either of these metrics. (D) Representative histograms of the blood velocities across the entire coronel hemisphere from an example young and aged mouse, demonstrating a shift in the distribution shape. Across all mice in this study, we found a significantly increased right skewness (1.16 vs. 0.98, p = 0.002) in the distribution of blood velocities of the whole brain in the aged cohort versus the young cohort. ULM images were rendered using MATLAB (R2019a, https://www.mathworks.com/).