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. 2022 Jul 15;13:892701. doi: 10.3389/fimmu.2022.892701

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Copyright © 2022 Sampath, Natarajan, Moideen, Kathamuthu, Hissar, Dhanapal, Jayabal, Ramesh, Tripathy, Ranganathan, Babu and Bethunaickan

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Representative gating strategy for monocytes and their subsets. Monocyte populations were gated on whole blood cell plot by Forward (FSC-A) and Side scatter (SSC-A) based on their size and granularity. Singlets were discriminated against doublets by FSC (A) vs FSC (H). Dump gate were used to select lineage negative cells by excluding T cells, B cells and NK cells by CD2, CD3, CD19 and CD56 markers. Total monocytes were gated using CD14 and CD16 surface expression. HLA-DR+ and CD16+ cells were gated to obtain the pure monocyte population which were further gated using CD14 and CD16 surface markers to distinguish classical (HLADR⁺CD14⁺⁺CD16^-), intermediate (HLADR⁺CD14⁺⁺CD16⁺) and non-classical (HLADR⁺CD14^+/-CD16⁺) monocytes.