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. 2024 Jun 3;16(7):1495–1514. doi: 10.1038/s44321-024-00084-4

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© The Author(s) 2024

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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) Schematic representation of the screening platform, including 7-color 3D imaging and STAPL-3D single organoid segmentation and quantification for target prioritization. (B, C) Representative single channel and merged 3D multispectral images of the segmented data, showing heterogeneous expression of selected FGS probes between PDO lines. Scale bars 50 μm. (D, E) Quantification of percentage positively stained organoids (dot size) and normalized fluorescent intensity (blue-to-red color gradient) for the two PDO biobanks and healthy tissue control organoids. (B–E) n = 3 (NB) and n = 4 (BC) independent experiments. Source data are available online for this figure.