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Table 1.

Immunophenotyping by mIF using two OPAL panels.

Phenotype	Biomarker expression
Opal Panel 1
Epithelial cells	(CK⁺PD-L1⁺) + (CK⁺)
Epithelial cells PD-L1⁺	CK⁺PD-L1⁺
Total T-cell lymphocytes	(CD3⁺ only) + (CD3⁺CD8⁺PD1⁻) + (CD3⁺PD1⁺) + (CD3⁺CD8⁺PD1⁺)
Cytotoxic T cells	(CD3⁺CD8⁺PD1⁻) + (CD3⁺CD8⁺PD1⁺)
T cells, antigen experienced	(CD3⁺PD1⁺) + (CD3⁺CD8⁺PD1⁺)
Total macrophages	(CD68⁺PD-L1⁻) + (CD68⁺PD-L1⁺)
Macrophages PD-L1⁺	CD68⁺PD-L1⁺
Opal Panel 2
Total B-cell lymphocytes	CD20⁺
CD45RO⁺ cells	CD45RO⁺
Granzyme B⁺ cells	Granzyme B⁺
FOXP3⁺ cells	FOXP3⁺
CD57⁺ cells	CD57⁺

Phenotype	Biomarker expression
Opal Panel 1
Epithelial cells	(CK⁺PD-L1⁺) + (CK⁺)
Epithelial cells PD-L1⁺	CK⁺PD-L1⁺
Total T-cell lymphocytes	(CD3⁺ only) + (CD3⁺CD8⁺PD1⁻) + (CD3⁺PD1⁺) + (CD3⁺CD8⁺PD1⁺)
Cytotoxic T cells	(CD3⁺CD8⁺PD1⁻) + (CD3⁺CD8⁺PD1⁺)
T cells, antigen experienced	(CD3⁺PD1⁺) + (CD3⁺CD8⁺PD1⁺)
Total macrophages	(CD68⁺PD-L1⁻) + (CD68⁺PD-L1⁺)
Macrophages PD-L1⁺	CD68⁺PD-L1⁺
Opal Panel 2
Total B-cell lymphocytes	CD20⁺
CD45RO⁺ cells	CD45RO⁺
Granzyme B⁺ cells	Granzyme B⁺
FOXP3⁺ cells	FOXP3⁺
CD57⁺ cells	CD57⁺