File:Human-T-Lymphotropic-Virus-1-Visualized-at-the-Virological-Synapse-by-Electron-Tomography-pone.0002251.s001.ogv

English: Tomographic reconstruction showing a large number of HTLV-1 particles of variable size in the space between a chronically infected MS9 cell (top) and a potential Jurkat target cell (bottom). The particles are characterized by an electron-dense core surrounded by a less dense area and enveloped by the electron-dense viral membrane. The average diameter of HTLV-1 particles derived from MS9 cells was 126.0 +/− 31.3 nm (SD). White arrows indicate areas in the MS9 cell where viral capsid protein accumulates below the membrane in anticipation of virus budding. Three small parts of mitochondria (upper left corner) and several microtubules are visible in the MS9 cell cytoplasm. A slice through the entire tomogram is shown in Figure 3D. The sample was fixed with PFA/glutaraldehyde, embedded in Agar 100 resin, post-fixed with OsO4 and stained with lead citrate. The reconstructed 3D volume is composed of tomograms obtained from four serial sections. The tomograms were calculated from tilt series of 121 images (tilt range −60° to +60°, 1° increment). The images were recorded with a 2k×2k Gatan CCD camera (pixel size 1.21 nm) at an under focus of −0.2 µm with an FEI Tecnai F30 FEG-TEM (300 kV) in Boulder, Colorado using the software SerialEM. The reconstruction was calculated using the software IMOD. The tomogram represents a volume with an area of 1.94×1.28 µm2 and a thickness of 640 nm. The scale bar at the end of the movie corresponds to 400 nm.