Table 4. Comparison between instruments of the increment in average cell frequencies obtained after staining with the panels components.
| Antibodies | Reference sample I | Reference sample II | Reference sample III | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Canto | Fortessa | MQ | p- value | Canto | Fortessa | MQ | p- value | Canto | Fortessa | MQ | p- value | |
| CD45 VioGreen | 98.3 | 97.2 | 98.3 | ns | 99.5 | 94.0 | 98.5 | ns | 97.0 | 97.0 | 99.0 | ns |
| CD3 VioBlue | 75.6 | 73.0 | 76.7 | ns | 75.3 | 73.4 | 77.5 | ns | 67.2 | 69.0 | 71.7 | ns |
| CD14 VioBlue | 1.13 | 2.5 | 1.0 | ns | 1.11 | 3.7 | 1.0 | ns | 1.7 | 12.92 | 1.1 | ** |
| CD19 VioBlue | 16.0 | 16.4 | 14.5 | ns | 7.7 | 7.7 | 12.5 | ns | 4.19 | 9.76 | 20.2 | * |
| Sytox® Blue | 17.0 | 12.4 | 18.5 | ns | 16.2 | 13.1 | 14.5 | ns | 13.6 | 12.3 | 19.0 | ns |
| CD56 APC-Vio770 | 2.9 | 2.5 | 2.4 | ns | 2.2 | 3.0 | 2.7 | ns | 9.29 | 9.8 | 8.3 | ns |
| CD3 PerCP-Vio700 | 76.3 | 74.0 | 77.5 | ns | 74.6 | 77.5 | 77.4 | ns | 68.4 | 69.7 | 72.1 | ns |
| TCRγδ PerCP-Vio700 | 1.9 | 2.5 | 2.9 | ns | 1.7 | 2.7 | 3.23 | ns | 2.6 | 2.5 | 2.9 | ns |
| TCRγδ VioBlue | 1.6 | 1.6 | 2.4 | ns | 1.7 | 1.4 | 2.71 | ns | 2.5 | 2.7 | 2.4 | ns |
| CD56+CD16 APC | 33.0 | 28.0 | 31.6 | ns | 33.4 | 30.0 | 33.7 | ns | 36.0 | 34.5 | 31.5 | ns |
| CD56+NKG2A PE-Vio770 | 59.0 | 63.4 | 57.5 | ns | 59.6 | 63.8 | 61.0 | ns | 48.8 | 51.6 | 62.9 | ns |
| CD56+NKG2C PE | 6.3 | 6.2 | 6.4 | ns | 7.9 | 4.9 | 6.0 | ns | 11.0 | 10.7 | 12.7 | ns |
| CD56+NKG2D PerCP-Cy5.5 | 92.6 | 91.3 | 92.9 | ns | 90.8 | 94.9 | 92.8 | ns | 97.8 | 95.5 | 93.5 | ns |
| CD56+PanKIR2D FITC | 35.1 | 33.9 | 36.0 | ns | 34.2 | 33.9 | 29.2 | ns | 18.1 | 19.3 | 21.4 | ns |
| CD56+CD25 VioBrightFITC | 7.2 | 7.8 | 8.0 | ns | 6.9 | 7.2 | 7.4 | ns | 22.3 | 23.4 | 25.2 | ns |
| CD56+NKp44 PE-Vio770 | 1.0 | 0.8 | 1.2 | ns | 5.6 | 4.4 | 6.6 | ns | 6.1 | 6.0 | 7.5 | ns |
| CD56+CD107a PE | 23.7 | 24.2 | 19.5 | ns | 24.0 | 25.2 | 26.3 | ns | 31.2 | 28.2 | 33.8 | ns |
Three reference samples (Reference sample I–III) were analysed using three different flow cytometers at three different centers by independent operators using an optimized protocol. The performance of the backbone antibodies (white shades) was evaluated using single stain controls for each measured parameter with background subtracted from corresponding negative controls. The drop-in markers for NK phenotype panel (Bold) were analysed, when gated on NK cells (CD45+CD3−CD56+). Background staining in NK phenotype panel drop-in channels were eliminated using appropriate fluorescence minus one (FMO) controls. For the NK function panel drop-in markers; CD107a, CD25 and NKp44, NK cells were stimulated with A431 cells and unstimulated values were subtracted before analysis (Italics). The values shown for each parameter correspond to the average frequency of stained cells from two technical replicates measured in triplicates, following subtraction of appropriate background signal. The statistical comparisons were performed using non-parametric Kruskal-Wallis. Only significant p-values (<0.05) are shown.