Video 8. CLSM on macrophage and neutrophil behavior at the exponential phase of Mm infection.

Double transgenic Tg(mpeg1:EGFP) and Tg(lyz:DsRed2) larvae infected with Mm were imaged alive every ~60 sec at 3 dpi for 2.5 hours by CLSM (Nikon A1) before fixation. The two infected GFP-positive (green) macrophages and two infected DsRed-positive (blue) neutrophils that were correlated with 3D block-face SEM images are indicated by arrows. The indicated macrophages (MP1 and MP2) phagocytized Mm approximately 2 and 1.5 hours before fixation respectively and these cells remained GFP-positive after fixation. The neutrophil (NP1) containing Mm was observed 2.5 hours before fixation and undergoes cell death, observed as rapid (within ~1 min) disappearance of the fluorescent signal, ~30 min before fixation. The second neutrophil (NP2) was recruited to another infected neutrophil ~30 min before fixation. The maximum intensity projection from 11 CLSM images (step size in z-direction, 5.56 µm) is shown at 10 frames per second. Scale bar: right panel 100 µm and for the left panel 50 µm.