Video 10. Three-dimensional block-face SEM images of necrotic neutrophil undergoing netosis.

A double transgenic Tg(mpeg1:EGFP) and Tg(lyz:DsRed2) larva infected with M. marinum was fixed and block face SEM imaging was performed on an 80 by 80 µm region of interest using a Quanta FEG 250 with a Gatan 3View Ultramicrotome. The video shows 70 images in z-direction (23 nm pixel size) of the neutrophil (NF2, Fig. 4). Note the partial intact nuclear envelope (arrow) and the vesicles (arrowheads) fusing with bacterial aggregate. The images were acquired with every 150 nm step size in z-direction and are in this video visualized at 10 frames per second. Scale Bar: 2 µm.