Fig. 4
Immune cell dynamics in different bone types show comparable motility patterns. a 3D fluorescence images acquired by LIMB in the femoral marrow of a CD19:tdRFP mouse at day 7, day 14, day 28, and day 60 after surgery. Mature B lymphocytes express tdRFP and are displayed in green, whereas the vasculature was labeled with Qdots and is displayed in red. Scale bar = 50 µm. All images are snapshots of 45 min movies, with images acquired every 30 s. The movies are provided as Supplementary Material. b Time-lapse 3D fluorescence image acquired by LIMB in the bone marrow of a CD19:tdRFP mouse at day 90 post-surgery. The tracks of the B lymphocytes smaller than 500 µm3 (defined as B cells with a maximum diameter of 10 µm) are shown in cyan, whereas those with a volume larger than 500 µm3 (defined as plasma cells) are shown in violet. Scale bar = 30 µm. c Similar to b, time lapse 3D image of the bone marrow of a CD19:tdRFP mouse with corresponding tracks of B and plasma cells in the calvarium and d the tibia. Scale bar = 30 µm. Representative movies with cell motility tracks for LIMB, calvarial, and tibial imaging are provided as Supplementary Material. e Quantification of cell volumes, mean velocities, and displacement rates of B lymphocytes from movies acquired by LIMB (n = 4 mice), within the calvarial bone (n = 3 mice), and the tibia (n = 2 mice). Similar cell subset frequencies and mean velocities of B lymphocytes were measured by LIMB in the femoral bone marrow, by calvarial imaging as well as by tibial imaging. We statistically analyzed the data in e using t-test (*p < 0.05; **p < 0.01; ***p < 0.001)
