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. 2023 Oct 9;13:16994. doi: 10.1038/s41598-023-43435-w

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© The Author(s) 2023

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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Staining intensity evaluation over 20 iterations of seqIF protocol. (a) Epitope stability of four markers targeting different cell types and cellular localization (CD45, FoxP3, CK, and Vimentin) quantified after 5 cycles, 10 cycles, 15 cycles and 20 cycles of elution compared to the initial staining. All staining and elution steps were performed on the same tissue sample (human tonsil FFPE). The signal-to-background ratios (SBR) as a % of the initial staining are displayed on the right. Scale bar: 50 μm. n = 1. (b) Epitope stability of four markers targeting different cell types and cellular localization (CD45, FoxP3, CK, and Vimentin) quantified after 5 cycles, 10 cycles, 15 cycles and 20 cycles of elution compared to the initial staining. All staining and elution steps were performed on human lung adenocarcinoma (AC) and squamous cell carcinoma (SCC) FFPE samples. The SBR as a % of the initial staining are displayed on the right. Scale bar: 50 μm. n = 1.