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. 2020 Sep 1;10:14408. doi: 10.1038/s41598-020-71347-6

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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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Example of applying U-Net models to determine drug biodistribution in production run. (a–c), a FOV on proximal section of colon tissue obtained from mouse dosed with 3 mg/Kg αCDH-A647 for 24 h. (a1) Full color image of the FOV acquired by HCS (red: αCDH-A647; green anti-EpCAM; blue: DAPI). (a2) Nuclei identified by a U-Net model (yellow cross). (a3) Epithelial region segmented by another U-Net model (grey color zone). (b1) Identification of nucleated epithelial region in the FOV—yellow: nuclear areas approximated from detected nuclei; blue: segmented epithelial regions; red: selected nucleated epithelial regions by overlapping yellow and blue. (b2) Image of αCDH-A647 (red) with nuclei (blue) in the FOV. (c) Histogram of αCDH-A647 biodistribution in the FOV was obtained by overlapping αCDH-A647 in (b2) with nucleated epithelial regions in (b1). (d) Time courses of αCDH-A647 occupancy in nucleated epithelial cells of the whole colon proximal and small intestine duodenum tissue sections.