Background randomization of negative events improves visualization of high‐dimensional FACS data. Channel labels refer to the laser source (UV: ultraviolet V: violet B: blue YG: yellow/green R: red) and the median of wavelength detection by the bandpass filter. a) Background and b) positive signals across independent experiments as revealed by unstained and single‐peak rainbow beads, respectively. Machine performance across independent experiments.
Matrix overlay of antigen expression between run 1 and run 5.
Similar distribution of compensated data between independent experiments. The grey bar in the background represents the interquartile range of the distribution. Each dot represents a single independent experiment. b) Frequency of CD8 + T cells expressing a given antigen in 3 different healthy donors (HD).
a) Gating strategy of the identification of CD8 + T cells in peripheral blood used in this study. 27‐parameter FACS is reproducible across independent experiments.
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