Stimulated B cells show distinctive behavior compared with that of nonstimulated B cells. (A) Still images acquired at the indicated time points from an hour intravital imaging shows the localization of adoptively transferred nonstimulated (red; CMTMR) or LPS-stimulated B cells (green; CMFDA). The dimension of LN follicle was visualized by previously transferred B cells (blue; CMAC) and HEVs were visualized with by the intravascular injection of quantum dot 705 (purple). The stimulated B cells were cultured overnight in the presence of LPS (1 μg/mL) and transferred simultaneously with freshly isolated B cells. (B) The image shows displacements of nonstimulated (red color arrows) or LPS-stimulated B cells (green color arrows) from the HEVs. The HEVs are highlighted (light purple) using the surfaces function in the Surpass view of Imaris. The tracks were randomly selected from tracks having durations greater than 10 minutes in an hour-long video. The individual tracks were analyzed to assess displacement and speed variability. Time for TEM of individual cells measured between images acquired 5 to 65 minutes after transfer (*P < .05; ***P < .0001). (C) Most LPS-stimulated B cells enter into B-cell follicles by 3 hours after transfer. The image shows the distribution of transferred B cells 3 hours after transfer. Pretransferred B cells (blue color spot) delineate the LN follicle. Locations of nonstimulated B cells (red), stimulated B cells (green) shown as colored balls. The HEVs are delineated by a light purple color. The graph shows the distance between each cell and a designated center of the LN follicle at various time points after cell transfer. The black bar is the mean value of each group. Scale bars are 50 μm. This experiment is representative of 3 performed.