Objective

Purpose: Cytometry is a powerful analytical tool for the analysis of multiple biological parameters within heterogeneous cell populations. Here we present validation data on three assays (regulatory T-cells (Treg), T-helper 17 cells (Th17) and plasma cell assays).

Methods: Human PBMC samples from 5 healthy donors were loaded in microfluidic cassettes (ZellSafe™) and assays were validated for Treg, Th17 and plasma cells. Data processing was performed using an automated gating algorithm.

Results: All three assay panels were developed using commercially available antibodies. Determination of the intra-assay precision showed a mean CV of 16.2 % (Th17 assay), 19.1 % (Treg assay) and 14.6% (plasma cell assay). The intermediate CV, which comprises both within- and between-batch variability was 19.6 % (Th17 assay), 22.5 % (Treg assay) and 19.3 % (plasma cell assay). Stability analysis of 5 PBMC samples from 5 donors, stored in ZellSafe™ cassettes at 4°C for 3 months, revealed that 5/5 samples (Th17 and Treg assay) and 4/5 samples (plasma cell assay) were within the 25 % range from baseline level,  which is within predefined acceptance criteria .

Conclusion: Three Chipcytometry assays for quantification of Treg, Th17 and plasma cells in human PBMC samples were successfully validated.