Figure 4
From: Performance of plate-based cytokine flow cytometry with automated data analysis
Effect of overnight "resting" on cryopreserved PBMC. (A) Representative comparison of fresh PBMC, cryopreserved PBMC, and cryopreserved PBMC incubated overnight prior to stimulation (all from the same CMV+ donor). Unstimulated samples (top row) and CMV pp65 peptide mix-stimulated samples (bottom row) show similar results, but the IFNγ fluorescence intensity (MFI) is greatest in the cells rested overnight prior to stimulation. (B) Results of triplicate samples stimulated with CMV pp65 peptide mix and analyzed for percentage of IFNγ+ cells or IFNγ mean fluorescence intensity. Error bars represent SEM. All data were gated on CD3+CD4- cells. Similar results were obtained for TNFα+ cells (not shown). Results are representative of two similar experiments.