Table 4.
Example spillover spreading matrix for a 22-color experiment (SSM). Columns are to be interpreted as the detectors that are receiving spreading error. The value of the spreading error is relative and additive, i.e., a detector that is receiving spreading error from several fluorochromes is overall showing a higher loss in resolution. Examples as to how the SSM can be utilized during panel design are described in “experimental workflow”
| fluorochrome╲detector | B-530 (FITC) |
B-710 (PerCPCy5.5) |
R-670 (APC) |
R-710 (AF700) |
R-780 (APC-Cy7) |
U-379 (BUV395) |
U-570 (BUV563) |
U-610 (BUV610) |
U-660 (BUV661) |
U-740 (BUV737) |
U-800 (BUV805) |
V-450 (BV421) |
V-470 (BV510) |
V-570 (BV570) |
V-610 (BV605) |
V-655 (BV650) |
V-710 (BV711) |
V-780 (BV785) |
YG-586 (PE) |
YG-610 (Pe-Tx) |
YG-670 (Pe-Cy5) |
YG-780 (Pe-Cy7) |
total SE contributed |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| FITC | 0 | 0.599 | 1.07 | 0 | 0 | 0 | 0 | 0.356 | 0 | 0.729 | 0 | 0 | 0.396 | 0.449 | 0.455 | 0.386 | 0.24 | 0 | 1.37 | 1.06 | 0 | 0.296 | 7.406 |
| PerCPCy5.5 | 0.247 | 0 | 2.24 | 4.18 | 1.91 | 0.223 | 0 | 0 | 0.275 | 2.92 | 1.76 | 1.72 | 0.465 | 0 | 4.92E-03 | 0.698 | 1.91 | 2.4 | 0 | 1.57 | 3.3 | 3.1 | 28.92292 |
| APC | 0 | 1.09 | 0 | 2.78 | 1.29 | 0.0662 | 0.0775 | 0 | 0.268 | 0.889 | 0.486 | 0.361 | 0 | 0 | 0.183 | 0.905 | 0.55 | 0.433 | 0.811 | 0.391 | 4.28 | 1.23 | 16.0907 |
| AF700 | 0.132 | 0.5 | 0.767 | 0 | 2.18 | 0.119 | 0.139 | 4.87E-04 | 0 | 1.62 | 0.738 | 0.44 | 0.143 | 1.99E-03 | 0 | 0.143 | 0.368 | 0.737 | 1.29 | 0.907 | 0.444 | 1.77 | 12.439477 |
| APC-Cy7 | 0.0822 | 0.231 | 0.847 | 0.639 | 0 | 0.0796 | 0.125 | 0.0848 | 0.132 | 0.747 | 2.06 | 0.974 | 0.0922 | 0 | 0.0874 | 0.196 | 0.142 | 1.33 | 0 | 0 | 0.507 | 5.49 | 13.8462 |
| BUV395 | 0.896 | 0 | 0 | 0 | 0.861 | 0 | 0 | 0 | 0 | 0 | 0 | 0.624 | 0 | 1.39 | 0 | 0 | 0 | 0.709 | 0 | 0 | 1.5 | 0 | 5.98 |
| BUV563 | 0.375 | 1.33 | 2.55 | 0 | 0 | 2.01 | 0 | 1.26 | 0.853 | 2.32 | 1.29 | 0.602 | 0.492 | 2.66 | 1.39 | 0.739 | 0.53 | 0.302 | 22.3 | 11.7 | 3.23 | 1.2 | 57.133 |
| BUV610 | 0 | 2.22 | 3 | 1.15 | 0.442 | 1.34 | 0 | 0 | 0.781 | 3.34 | 1.97 | 0.517 | 4.11E-03 | 1.27 | 1.4 | 1.03 | 1.01 | 0.63 | 8.39 | 18.8 | 5.12 | 2.43 | 54.84411 |
| BUV661 | 0 | 1.66 | 12.9 | 6.44 | 3.07 | 0.953 | 0 | 0.535 | 0 | 7.07 | 3.77 | 0.627 | 0.565 | 0.533 | 0.627 | 1.9 | 0.979 | 1.11 | 0 | 1.91 | 5.4 | 2.68 | 52.729 |
| BUV737 | 0 | 0.519 | 0 | 1.72 | 1.22 | 0.182 | 0 | 0.194 | 0 | 0 | 2.17 | 0.347 | 0 | 0.242 | 0 | 0 | 0.337 | 0.605 | 1.97 | 1.07 | 0.649 | 0.816 | 12.041 |
| BUV805 | 0.237 | 0 | 0.513 | 0 | 1.07 | 0.967 | 0 | 0.165 | 0 | 0.641 | 0 | 0.381 | 0 | 0.294 | 0.165 | 0.175 | 0 | 0.549 | 0.0998 | 0 | 0 | 0.849 | 6.1058 |
| BV421 | 0 | 4.24E-03 | 0.738 | 0 | 0 | 0.123 | 0 | 0 | 0 | 0.576 | 0.19 | 0 | 0.632 | 0.182 | 0.214 | 0.157 | 0.132 | 0.118 | 0.264 | 0 | 7.97E-03 | 0.175 | 3.51321 |
| BV510 | 0.207 | 0.235 | 0 | 5.15E-03 | 0.193 | 0.13 | 0.153 | 0.148 | 0.224 | 0 | 0 | 0.563 | 0 | 1.39 | 0.952 | 0.627 | 0.284 | 0.313 | 1.51 | 0.865 | 0 | 0.176 | 7.97515 |
| BV570 | 0.205 | 0.749 | 1.27 | 0.534 | 0 | 0 | 0.223 | 0.485 | 0 | 1.37 | 0.416 | 1.48 | 0.386 | 0 | 2.76 | 2.03 | 0.695 | 0.761 | 7.75 | 4.02 | 1.59 | 0.794 | 27.518 |
| BV605 | 0 | 0.927 | 2.23 | 0.449 | 0.466 | 3.11E-03 | 0 | 0.81 | 0.283 | 2.04 | 0.971 | 1.35 | 0.396 | 2.61 | 0 | 3.22 | 1.27 | 1.42 | 4.74 | 5.48 | 2.42 | 1.36 | 32.44511 |
| BV650 | 0 | 0.889 | 0 | 1.38 | 0 | 0 | 1.15 | 0 | 1.18 | 3.71 | 1.28 | 1.36 | 1.15 | 1.64 | 1.12 | 0 | 1.5 | 1.65 | 0 | 0 | 0 | 1.57 | 19.579 |
| BV711 | 0.839 | 1.61 | 1.27 | 3.72 | 1.86 | 0.784 | 1.1 | 0.584 | 0.638 | 3.77 | 3.15 | 2.54 | 0.914 | 1.32 | 0.62 | 0.655 | 0 | 3.59 | 0 | 3.28 | 0 | 1.85 | 34.094 |
| BV785 | 0 | 0.389 | 1.05 | 0.608 | 0.809 | 0.316 | 0 | 0.349 | 0 | 1.94 | 2.47 | 3.22 | 0.641 | 0 | 0.349 | 0.544 | 0.567 | 0 | 0 | 2.03 | 0.834 | 0.62 | 16.736 |
| PE | 0.144 | 0.505 | 0.688 | 0.129 | 0 | 0.0902 | 0 | 0.153 | 0.161 | 0.505 | 0.18 | 0.338 | 0.113 | 1.44 | 0.81 | 0.465 | 0.239 | 0.225 | 0 | 3.32 | 1.16 | 0.382 | 11.0472 |
| Pe-Tx | 0 | 1.09 | 0.552 | 0.455 | 0.124 | 1.01E-03 | 0 | 0.254 | 0.122 | 0.513 | 0.259 | 0.249 | 0.0573 | 0.511 | 0.736 | 0.597 | 0.531 | 0.282 | 3.4 | 0 | 2.06 | 1.08 | 12.87331 |
| Pe-Cy5 | 0.0981 | 2.05 | 2.58 | 1.68 | 0.695 | 0.0638 | 0 | 0.101 | 0.0805 | 0.885 | 0.409 | 0.305 | 0.0746 | 0.158 | 0.151 | 0.681 | 1.07 | 0.446 | 1.52 | 0.683 | 0 | 2.07 | 15.801 |
| Pe-Cy7 | 0.207 | 0.346 | 0.444 | 0.257 | 0.559 | 0.187 | 0.306 | 0 | 0 | 0.721 | 0.874 | 0.629 | 0.225 | 0 | 0 | 0.225 | 0 | 0.865 | 0 | 1.25 | 0.489 | 0 | 7.584 |
| total SE collected | 3.6693 | 16.94324 | 34.709 | 26.12615 | 16.749 | 7.63792 | 3.2735 | 5.479287 | 4.9975 | 36.306 | 24.443 | 18.627 | 6.74621 | 16.09099 | 12.02432 | 15.373 | 12.354 | 18.475 | 55.4148 | 58.336 | 32.99097 | 29.938 |