MCL#2 Fortessa fcs files

After overnight recovery, purified T cells were stained with LIVE/DEAD Zombie fixable dead cell stain (0.2% in PBS) at RT in the dark for 15 minutes. T cells were washed twice in FACS buffer (PBS + 2% FCS) and stained in FACS buffer and 1% human Fc block (BD Pharmingen Human BD Fc Block) with mAb for the T cell surface markers CD4 BUV 395, CD8 BUV 805, CCR7 PE and CD45RA PerCP Cy5.5 (Table 1) on ice for 30 minutes. Single colour compensation and fluorescence minus one (FMO) control samples were prepared for CCR7 and CD45RA. CD19+CD20+ target cell lines (Raji, Mino, JeKo-1) were labelled with CTV and then stained with 0.2% LIVE/DEAD Zombie fixable dead cell stain, as above. T cells and target cells were washed twice after mAb staining in FACS buffer, resuspended at 2x106 cells/ml in TCM, and co-incubated at a ratio of 1:1 +/- blinatumomab (10 ng/ml) for 1 hour at 37°C in a 5% CO2 incubator. The samples were vortexed vigorously for 10 seconds and fixed by the addition of an equal volume of PBS containing 4% paraformaldehyde (Electron Microscopy Sciences, USA). Samples were acquired on a 5 laser Becton Dickinson LSR Fortessa (355, 405, 488, 561, 633 nm) conventional flow cytometer (BD Biosciences, USA). CS&T bead quality control testing was performed daily. Analysis was performed using FlowJo software (v10.8.1, BD Bioscience, USA).