Figure 1.

Consequential gating strategy for acquiring samples and detecting DEs, Bcon, and Tcon by using FACS and FlowJo

(A) Dot plots show how to sequentially gate live single lymphocytes and exclude doublets and dead cells in your PBMC sample.

(B) Left dot plot shows how to use a 2-parameter plot of CD19 versus CD5 to divide gated lymphocytes into B_con (CD5⁻CD19⁺); T_con (CD5⁺CD19⁻), and CD5⁺CD19⁺ populations. Right dot plots show how to analyze each gated subpopulation for DEs as identified by the coexpression of TCRαβ and BCR. Note: most DEs are located within the CD5⁺CD19⁺ compartments, rare among B_con, and virtually absent among T_con. Consequently, analysis and sorting of DEs, is limited to DEs localized within the CD5⁺ CD19⁺ population. Numbers indicate percentages in each quadrant. This figure is adopted from the manuscript by Ahmed and colleagues (Ahmed et al., 2019).