Data Supplement

Files in this Data Supplement:

• Video Legend (PDF, 31.2 Kb)
  - Legend for video and detailed MIAME array description.
• Video (MOV, 1.18 Mb)
  - Movie of FLU+ CD8+ cells interactions with B cells. Tetramer Flu+/CD8+ cells after 9 days of IVS, and fresh autologous CD19+ cells were labeled with tFlu, CD8 and CD19 antibodies. Cells were sorted by high-speed flow cytometry, mixed at 1:1 ratio in a poly-D-lysine/fibronectin coated gridded culture dish (MatTek Corporation), and cultured o.n. in 10% human serum Iscove’s medium, supplemented with 300 IU/ml rIL2, and DAPI. The cells were incubated at 37oC in 5%CO2 using an automatically controlled environmental chamber (Incubator XL) mounted on an Axiovert 200M inverted microscope (Zeiss) and video-recorded during o.n. culture. The movie clip shows a 2.5-hour sequence of snapshots captured at 5-minute intervals using a 25X Plan-Neofluar (Phase-2, multi-immersion) objective under phase-contrast illumination of the Axiovert 200M inverted microscope (Zeiss). The original images were automatically acquired at 1,344 x 1,024 pixel resolution and 12-bit grayscale depth using an ORCA-ER CCD camera (Hamamatsu) controlled by a customized OpenLab time-lapse imaging automation (Improvision). The final images depicting the field of interest over the indicated time frame were cropped at 596 x 716 pixel resolution and the movie clip was exported using a Sorenson video compression method into a QuickTime format. The full 15-hour video over the entire imaged field at the original pixel resolution is available upon request. Keys: Fluorescent labels: green (AlexaFluor 488) = CD19; blue (AlexaFluor 647) = CD8; red (PE) = FLU M1 iTAg MHC Tetramer; yellow = DAPI.