TABLE 1.
Antibody-Opal pairing strategy.
| Opal | Opal brightness rankings | Spectral bleed | Initial pairing |
|---|---|---|---|
| Opal 520 | Highest | PD1/CD4/ICOS | |
| Opal 540 | Medium | CD20 | |
| Opal 570 | Medium | Ki67/FOXP3 | |
| Opal 620 | Medium | PD1/CD4/ICOS | |
| Opal 650 | Highest | PD1/CD4/ICOS | |
| Opal 690 | Lowest | Ki67/FOXP3 |
The table shows the strategy used for antibody-Opal pairing. The co-localized surface markers (PD-1, CD4 and ICOS) were associated with the brightest Opals using the rankings on the Vectra Polaris scanner. The markers not expressed in the same cellular compartment were associated with Opals that are more subject to spectral bleed with the Vectra Polaris scanner (black boxes).