ENDS formation in a flow chamber. Human neutrophils labeled with CD16-AF647 antibody rolled on E-selectin substrate under perfusion with Ca2+/Mg2+ containing HBSS buffer. Flow direction is flow left to right. WSS was increased with 10 dyn/cm2 increments to 50 dyn/cm2. Image segmentation was applied to quantify ENDS formation; cyan – cell body, yellow – tethers that are still connected to the cell body, magenta – ENDS that are not connected to the cell body. Original speed, 3.75 frames/s; every fifth frame exported at 10 frames/s; playback speed is 13 times faster than real speed.
