Four color facscalibur 2 double laser flow cytometer

For identification of lymphocyte subsets, 50 μl of whole blood was incubated with fluorochrome-labeled monoclonal antibodies against CD3, CD4, CD8, CD45, (Cat. no. 342417) CD19, and CD16 + CD56 (Cat. no 342416) (all from BD Biosciences). After vortexing and incubation in the dark at room temperature for 15 min, 500 μl BD FACS Lyse solution (1:10) was added. The tubes were then vortexed and incubated in the dark at room temperature for 15 min. Finally, all samples were evaluated with four-color FACSCalibur II double-laser flow cytometer (BD Biosciences). At least 100,000 events were analyzed in the initial FSC/SSC dot plot.

To calculate the absolute count of lymphocyte subsets, 50 μl of whole blood was incubated with fluorochrome-labeled monoclonal antibodies against CD3, CD4, CD8, CD45 (Cat. no. 342417), CD19, and CD16+CD56 (Cat. no 342416) (all from BD Biosciences). After vortexing and incubation in the dark at room temperature for 15 minutes, 500 µl BD FACS Lyse solution (1:10) was added. The tubes were then vortexed and incubated in the dark at room temperature for 15 minutes. Finally, all samples were evaluated with four-color FACSCalibur II double-laser flow cytometer (BD Biosciences). At least 100,000 events were analyzed in the initial FSC/SSC dot plot.