Table 1.
Virtual Quadruple Staining Sequence for the Detection of ILC1, ILC2, or ILC3.
| Staining 1 |
Staining 2 |
Staining 3 |
Staining 4 |
|||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Cell Types to Be Detected | Primary Antibody (Species) | Secondary Antibody–Conjugate >Chromogen | Primary Antibody (Species) | Secondary Antibody– Conjugate >Chromogen |
Primary Antibody (Species) | Secondary Antibody–Conjugate >Chromogen |
Primary Antibody (Species) | Secondary
Antibody–Conjugate >Chromogen |
||||
| ILC1 Th1/Tc1 |
T-bet (rabbit) |
anti-rabbit–HRP >NovaRED |
Digital scanning/stripping immune complexes and dye | CD56 (rabbit) |
anti-rabbit–HRP >NovaRED |
Digital scanning/stripping immune complexes and dye | CD3 (rabbit) |
anti-rabbit–HRP >NovaRED |
Digital scanning/stripping immune complexes and dye | CD20/CD79α (mouse) |
anti-mouse–HRP >NovaRED |
Digital scanning/composition of virtual 4-color image |
| ILC2 Th2/Tc2 |
GATA3 (rabbit) |
anti-rabbit–HRP >NovaRED |
CD56 (rabbit) |
anti-rabbit–HRP >NovaRED |
CD3 (rabbit) |
anti-rabbit–HRP >NovaRED |
CD20/CD79α (mouse) |
anti-mouse–HRP >NovaRED |
||||
| ILC3 Th17/Tc17 |
RORγt (mouse) |
anti-mouse–HRP >NovaRED |
CD56 (rabbit) |
anti-rabbit–HRP >NovaRED |
CD3 (rabbit) |
anti-rabbit–HRP >NovaRED |
CD20/CD79α (mouse) |
anti-mouse–HRP >NovaRED |
||||
Abbreviations: ILC, innate lymphoid cell; Th, helper T cell; Tc, cytotoxic T cell; HRP, horseradish peroxidase.