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. 2023 Mar 18;13:4487. doi: 10.1038/s41598-023-30286-8

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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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(a) Light microscopy of whole mount slides displaying H&E, TUNEL, and the DAB + channel respectively. Regions of interest were drawn (dashed curve) on H&E slides avoiding skin and muscle tissue. Color deconvolution separated the TUNEL stains into channels where the DAB + channel was isolated, and the percent positive staining can be calculated as the ratio of included pixels divided by the number of pixels in the ROI. (b) Magnified views (25X; 0.4 × 0.4 µm) of the highlighted regions (black box). Color deconvolution shows good agreement with the brown positive stain in TUNEL stain and absence of background stain. Scale bar represents 160 µm.