FIGURE 7.

Cell density and Opal expression quantification of drop control slides. (A) Cell density quantification of CD4⁺, ICOS⁺, PD-1⁺, and T_FH subpopulations (ICOS⁺CD4⁺PD-1⁺, CD4⁺PD-1⁺, ICOS⁺PD-1⁺, CD4⁺ICOS⁺) on monoplex slides, the mIHC slide and drop control slides without (w/o) ICOS, CD4, or PD-1. Each condition was quantified following the workflow shown in Figure 4A in a TLS GC (black dotted line in Figure 6). (B) Quantification of 520, 620, and 650 Opal mean expression (OME) on T_FH cells from an mIHC slide and drop control slides without (w/o) ICOS, CD4, PD-1 staining. Each condition was quantified followed the workflow as described above. The slides were scanned at ×20 magnification with the Vectra Polaris and the composite images were analyzed with InForm software (v.2.4.8).