Article

Three‐dimensional electron microscopy of entire cells

Swiss National Centre for Retroviruses, Zürich, Switzerland.
Journal of Microscopy (Impact Factor: 2.33). 02/1990; 157(Pt 1):115-26. DOI: 10.1111/j.1365-2818.1990.tb02952.x
Source: PubMed

ABSTRACT

The digital processing of serial electron-microscope sections containing laser-induced topographical references allows a three-dimensional (3-D) reconstruction of entire cells at a depth resolution of 40-60 nm by the use of novel image analysis methods. The images are directly processed by a video-camera placed under the electron microscope in TEM mode or by the electron counting device in STEM mode. The deformations associated with the cutting of embedded cells are back-calculated by new computer algorithms developed for image analysis and treatment. They correct the artefacts caused by serial sectioning and automatically reconstruct the third dimension of the cells. Used in such a way, our data provide definitive information on the 3-D architecture of cells. This computer-assisted 3-D analysis represents a new tool for the documentation and analysis of cell ultrastructure and for morphometric studies. Furthermore, it is now possible for the observer to view the contents of the reconstructed tissue volume in a variety of different ways using computer-aided display techniques.

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Available from: Michel Jourlin, Oct 22, 2014
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    • "A few years later Glaser and Glaser (1965) demonstrated a computer generated 3D-reconstruction from images derived from serial sectioning analyzed by a light microscope. Since these striking works many articles dealing with different kinds of 3D-reconstructions based on microscopical data (transmission electron microscope (Bron et al., 1990; Gremillet et al., 1991), lightmicroscopy of semithin-sections (Schmolke and Fleischhauer, 1984; Schmolke, 1996), lightmicroscopy of 5 to 50 μm thick sections have been published (Schormann and Zilles, 1998; Ourselin et al., 2001b). A disadvantage of sectioning, staining and mounting the sections on the slides is a lack of spatially perfect matching when the sections are superimposed. "
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    • "Elucidation of the relative positioning, shape and size of plant parenchyma cells in situ has hitherto been achieved by indirect methods involving microscopic analyses of thin, serial sections followed by stereological analysis (Considine, 1978; Considine, 1981; Cruz- Orive, 1997; Mùller et al., 1990; Underwood, 1970; Weibel, 1979; Weibel, 1980) or three-dimensional (3D) reconstruction of physical slices (Bron et al., 1990; Korn and Spalding, 1973; Korn, 1974; Lewis, 1926; Matzke, 1948; Williams, 1968). While appropriate for tissues composed of isodiametric cells, stereological methods underestimate mean cell sizes (Considine and Knox, 1981) and do not allow accurate determinations of intratissue variation (Parsons et al., 1989). "
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    • "Elucidation of the relative positioning, shape and size of plant parenchyma cells in situ has hitherto been achieved by indirect methods involving microscopic analyses of thin, serial sections followed by stereological analysis (Considine, 1978; Considine, 1981; Cruz- Orive, 1997; Mùller et al., 1990; Underwood, 1970; Weibel, 1979; Weibel, 1980) or three-dimensional (3D) reconstruction of physical slices (Bron et al., 1990; Korn and Spalding, 1973; Korn, 1974; Lewis, 1926; Matzke, 1948; Williams, 1968). While appropriate for tissues composed of isodiametric cells, stereological methods underestimate mean cell sizes (Considine and Knox, 1981) and do not allow accurate determinations of intratissue variation (Parsons et al., 1989). "
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