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. 2022 Feb 9;10:18. doi: 10.1186/s40478-022-01321-y

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Schematic overview and representative analyses of our multi-marker histological quantification approach based on scanned whole-slides. A Working flow of our experimental/analysis pipeline. B, C Representative quantification of cytoplasmic (B) and nuclear (C) immunohistochemical staining using single-cell segmentation performed with the HALO® software (Indica Labs, New Mexico, USA) on scanned serial sections. Left panels show whole-section scans of immunohistochemical staining. Right panels are enlarged views of the immunohistochemical staining (top), with the matching image segmentation (bottom). Nuclei are segmented in blue, and positive staining is segmented in red. Quantification results are shown in the tables associated, as follows: total number of cells (counted on the total number of nuclei on the whole scan), number of positive cells, and the percentage of positive cells over the number of total cells. Scale bar of the enlarged images = 50 µm. Abbreviations: formalin-fixed paraffin-embedded (FFPE), follicle-stimulating hormone (FSH), steroidogenic factor 1 (SF1)