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. 2018 Oct 12;8:15203. doi: 10.1038/s41598-018-33488-7

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© The Author(s) 2018

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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Sequential gating to identify immune populations. (A) All panels started from the identification of immune cells (CD45+) from the cell gate. (B) Panel II. Identification of neutrophils (high-SSC/CD45+/CD15+/CD14−) and macrophages (CD45+/CD15−/CD14+). (C) Two examples of the phenotypic identification of pro-inflammatory macrophages (CD11c+) from the macrophage gate. (D) Panel III. Identification of mast cells (CD117+/FcεRIα+) from a previous CD45+/CD14− gate (not showed). (E) Panel IV. Identification of T-helper (CD45+/CD3+/CD4+/CD8−) and T-cytotoxic (CD45+/CD3+/CD4−/ CD8+) from a previous CD45+/CD3+gate (not showed). (F) Panel IV. Identification of B-lymphocytes (CD45+/CD3−/CD19+) from a previous CD45+/CD3− gate (not showed).