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Figure 2 | Transplantation Research

Figure 2

From: Standardization of whole blood immune phenotype monitoring for clinical trials: panels and methods from the ONE study

Figure 2

Overview of the gating strategy for panel ONE 01: general immune phenotype, using the sample of a healthy individual. The data file of the stained lysed (EDTA spiked) whole blood (WB) was analyzed as follows: exclusion of non-single events (forward scatter time of flight versus forward scatter integral); gating of CD45+ leukocytes (anti-CD45 versus sideward scatter integral) – the counted CD45+ events were used as the reference for calculating the absolute cell number of indicated populations in WB; gating and exclusion of granulocytes (anti-CD45 versus sideward scatter integral); gating and exclusion of all CD14+ monocytes (anti-CD14 versus anti-CD64) – the gated CD14+ monocytes were used to further discriminate different inflammatory/differentiation stages of monocytes (anti-CD16 versus anti-CD14) resulting in CD14++CD16- classical monocytes, CD14++CD16+ and CD14+CD16++ monocytes, and anti-CD16 versus anti-CD64 to capture CD16+CD64+ monocytes; gating of lymphocytes (forward scatter integral versus sideward scatter integral); gating of CD56+NK cells, which were further subdivided into CD56dim and CD56highNK cells; gating of CD3+ T cells (anti-CD56 versus anti-CD3) – gated T cells were used for identification of CD4+ T-cells and CD8+ T-cells (anti-CD4 versus anti-CD8), and the gated lymphocytes were also used for identification of the B cell population (anti-CD19 versus anti-CD3). WB, whole blood.

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