How do you quantify reliability for your mass cytometry standard panel on PBMC samples?

We completed rigorous tests as part of our CLIA registration process. We analyzed peripheral blood mononuclear cells (PBMCs) from 9 healthy donors, and looked and four measures. For each measure, we set a coefficient of variation (CV) threshold, based on the literature. As a refresher, the coefficient of variation is simply the standard deviation divided by the mean. In general, a smaller CV is better.

• Inter-run: Same sample, different runs
  • 3.52% versus CV threshold of 25% criteria
  • one PBMC sample was divided into three replicates and running on CyTOF on the same day
• Intra-run: Same sample, same run
  • 6.66% versus 30% CV criteria
  • three replicates from one PBMC sample were run on different days
• Inter-operator: Same sample, different operators
  • 4.83%, 7.36% versus 25% CV criteria
  • two replicates processed by different operators were compared
• Stability: Same sample, across time
  • -0.01% versus +/- 25% change between days
  • four PBMC samples were split, stored, and analyzed on days 1 and 22

Want to read more?

Our full validation report also includes a description of all protocols and methods, assay design including step-by-step gating strategy, and more.