Digital three-dimensional visualization of cellular organelles studied by medium- and high-voltage electron microscopy.
Tilted thick sections (one-half to several micrometers) of biological specimens observed with medium- to high-voltage electron microscopes are extremely useful for the study of the three-dimensional (3-D) structure of organelles. If high resolution in 3-D visualization and 3-D reconstruction is needed, many images corresponding to various angles of rotation and tilt must be recorded. This necessitates very time-consuming work--including eventual photographic processing--before good positioning of the object is defined. We have developed software which permits very rapid and precise determination of the tilt-axis, the registration of tilted views, 3-D measurements and 3-D visualization. Images are digitized either from negative films or directly with a camera fitted to the microscope. The application of the software is performed in minutes and allows for a rapid check of the quality of the tilt-series and of the features of interest of the object. Application of the software to the study of the 3-D structure of active components of the nucleolus stained with silver is shown.
INSERM U 314, CHR Maison Blanche, Reims, France.
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Beorchia
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Ploton
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Menager
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Lebonvallet
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Thiry
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Bonnet
D002466:Cell Nucleolus / Q000648:ultrastructure*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D008854:Microscopy, Electron / Q000379:methods*; D012984:Software; D013692:Telophase