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. 2021 May 27;4:632. doi: 10.1038/s42003-021-02166-2

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© The Author(s) 2021

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Tissue samples were obtained from healthy and diseased tonsils (chronic tonsillitis) in 5-µm-thick formalin-fixed, paraffin-embedded samples. b The thin tissue sections were then stained with metal-conjugated antibodies and get ablated from the tissue’s surface through the argon plasma and analyzed by the time of flight mass spectrometer. c Individual marker images can be assembled and visualized using different colors to observe the marker’s coexpression and spatial organization. d Cells were segmented using CellProfiler software for single-cell quantification. e Single-cell analyses provide the marker expression distributions, including phenotype clustering based on the coexpression of markers. Phenographs of all normal and diseased ROIs show the different phenotypes that make up the sample to n = 25–27 groups. Immune and stromal markers were selected to cluster the entire dataset of the normal and diseased tonsil. The markers list includes CD20, CD68, CD3, CD4, CD8a, granzyme B, pankeratin, and E-cadherin. Created in Biorender.com.