Multicentre Harmonisation of a Six-Colour Flow Cytometry Panel for Naïve/Memory T Cell Immunomonitoring

Figure 5

Interoperator variability. Overall interoperator variability related to the identification of CD3, CD4, and CD8 major subpopulations and naïve/memory T cell subsets, by all operators, in cPBMC (upper panel) and WB (lower panel) samples. Interoperator variance was expressed as (a) CV calculated as the median of analyst-specific CV, derived from the three average donor triplicates for each marker; (b) bias, calculated with respect to the mean reference value (average of the values obtained from all the operator data centrally analysed by the ROP); and (c) ICC, as an index of reproducibility, calculated in a 2-way design with “analyst” and “donor” as random variables for each parameter, considering the mean value of donor triplicates. CV, bias, and ICC among sites are shown for local (red bar) and centralised (blue bar) data analyses.

(a)

(b)

(c)