Ca2+ transients in PMNs determined by motion-corrected two-photon lung intravital imaging of the Ca2+ indicator GCaMP6f (green) before, during, and after PMN transmigration in endothelial AJs of lung microvessels. PMN reporter was td-Tomato expressed under Ly6G promoter (red); endothelial junction labeled by anti-PECAM1 antibody (blue). Real-time data is supplied asVideo S1. Scale bars, 0.3 mm/pixel; image capture rate, 2 s/frame. The color scheme shows blue as the lowest and yellow as highest increase in cytosolic [Ca2+]. The dotted line shows the lung capillary wall with a monolayer of endothelial cells. Bv, blood vessel; Alv, alveolus. Time is in min and s. Scale bars, 5 m.
