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. 2020 Sep 10;10:14930. doi: 10.1038/s41598-020-71901-2

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© The Author(s) 2020

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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MC ex vivo verification using TPE-FLIM and bright field microscopy. Side by side comparison of TPE-FLIM image (mean fluorescence lifetime τ_m in the 300–1,300 ps range) (a) and corresponding brightfield microscopic image after staining of human skin biopsy cryo-sections for MCs with tryptase and counter stained nuclei with Mayer's hemalum solution (b) and a histogram of TPE-FLIM parameters measured for the 17 staining-proved MCs (nine MCs with a longer τ_m and eight MCs with a shorter τ_m) (c). The MCs ((1) and (2) in a and b) are marked with orange ellipses and the laser-burned labels (28 µm × 28 µm) are marked in red. Images have been rotated and zoomed to match the same orientation and size. Scale bar: 30 µm.