Figure S2 from Spectral Flow Cytometry Methods and Pipelines for Comprehensive Immunoprofiling of Human Peripheral Blood and Bone Marrow

Figure S2. Unmixing Accuracy Assessment. NxN permutations showing density plots of the same marker on the x-axis and every other fluorochrome plotted on the y-axis. Plots were manually examined for accuracy of unmixing. (A) T/B panel; (B) M/N/D panel; (E, F) BMC panel. The same PBMC donor sample was used for (A and B) and data represent cells gated as singlets, non-RBC, and live cells. (C) Density plot of CD159c-BYG575 vs CD19-BYG710 after automated unmixing. (D) Density plot of CD159c-BYG575 vs CD19-BYG710 after applying additional manual compensation of -2.79 using the SpectroFlo compensation tool. (F) NxN plots after removing antibody aggregates in NovaFluor Blue 610-70S – CD19. (G) Antibody aggregates are manually removed through the use of a NOT gate in the aberrant population.