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This variation poses a challenge for both pathologists\u2019 and AI-based downstream analysis. Minimizing stain variation computationally is an active area of research. To further investigate this problem, we collected a unique multi-center tissue image dataset, wherein tissue samples from colon, kidney, and skin tissue blocks were distributed to 66 different labs for routine H&amp;E staining. To isolate staining variation, other factors affecting the tissue appearance were kept constant. Further, we used this tissue image dataset to compare the performance of eight different stain normalization methods, including four traditional methods, namely, histogram matching, Macenko, Vahadane, and Reinhard normalization, and two deep learning-based methods namely CycleGAN and Pixp2pix, both with two variants each. We used both quantitative and qualitative evaluation to assess the performance of these methods. The dataset\u2019s inter-laboratory staining variation could also guide strategies to improve model generalizability through varied training data.<\/jats:p>","DOI":"10.1038\/s41598-026-40943-3","type":"journal-article","created":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T05:46:20Z","timestamp":1772084780000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Staining normalization in histopathology: Method benchmarking using multicenter dataset"],"prefix":"10.1038","volume":"16","author":[{"given":"Umair","family":"Khan","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jouni","family":"H\u00e4rk\u00f6nen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marjukka","family":"Friman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hesam","family":"Hakimnejad","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leena","family":"Latonen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teijo","family":"Kuopio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pekka","family":"Ruusuvuori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,2,26]]},"reference":[{"key":"40943_CR1","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1179\/his.2009.32.1.27","volume":"32","author":"NW Troiano","year":"2009","unstructured":"Troiano, N. 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