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. 2021 Mar 25;19:83. doi: 10.1186/s12951-021-00830-7

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© The Author(s) 2021

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — Gating strategies for the surface marker analysis of human peripheral blood mononuclear cells (PBMCs) (a) and macrophages (b). a PBMCs were first gated (black lined gate) on via sideward scatter area (SSC-A) and forward scatter area (FSC-A) on lymphocytes. Doublets were excluded by gating on forward scatter height (FSC-H) vs. FSC-A and then viable cells were gated by FSC-A vs. Live/Dead Marker. Thereafter, CD3 + CD45 + T cells were identified based on gating by CD3 vs. CD45, which were then separated into CD4 + and CD8 + T cells. Finally, the mean fluorescence intensity (MFI) of the marker of interest (filled grey histogram) was determined in comparison to the negative control staining (black lined histogram). b Macrophages were first gated via SSC-A vs. FSC-A to exclude cell debris from the analysis. Doublets were excluded by gating on FSC-H vs. FSC-A. Then, viable cells were identified (FSC-A vs. Live/Dead) and the MFI of the marker of interest (filled grey histogram) was determined in comparison to the negative control staining (black lined histogram)