FIG. 5.
Three-dimensional sectioning of MHV-infected cells and characterization of Hel-containing inclusions. Cells were imaged for Hel with the 488-nm laser to detect Cy-2. (A) Sagittal (xy) sections of the cells were obtained by z sectioning at 0.2-μm intervals, with resolution in the x, y, and z planes of 0.12 μm/pixel. The total z distance of 12 μm was imaged with 58 sections. The sections were transformed into a stack with NIH Image 1.62b. Section 1 was at the level of the coverslip, and section 58 was at the “top” of the cell most distant from the coverslip. The numbers in each panel are the numbers of the sections: 11 indicates 11 of 58. (B) Coronal (xz) reconstruction of a cell. The cell on the right of the panels in panel A was isolated, and the z stack was used to reconstruct the cell in the xz (coronal) plane with NIH Image 1.62b. Section 29 was used as the reference because it is in the middle of the cell in z dimension and shows the complexes clearly. The levels of the xz slices are shown by white lines 1 through 6, and the panels to the right show the respective slices. The viewpoint is indicated by the arrow, and the orientation of the left and right sides of the respective images is shown. The white line through the panels shows the level of section 29. (C) Axial (yz) reconstruction of the cell. This was performed as described for panel B, except in the yz plane.