[en] 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.
Arii T., Hama K. (1987) Method of extracting three‐dimensional information from HVTEM stereo images of biological materials. J. Electron Microsc. 36:177-195.
Bonnet N., Quintana C., Favard P., Favard N. (1985) Three‐dimensional graphical reconstruction from HVEM stereoviews of biological specimens by means of a microcomputer. Biol. Cell. 55:125-138.
Cataldo C., Souchier C., Stahl A. (1988) Three dimensional ultrastructure and quantitative analysis of the human Sertoli cell nucleolus. Biol. Cell. 63:277-285.
Franck J. (1980) The role of correlation techniques in computer image processing. Computer Processing of Electron Micrographs , ed. by, P. W. Hawkes, Springer‐Verlag, Berlin; 187-222.
Géraud G., Soyer A., Epelboin Y. (1988) Three‐dimensional computer reconstruction from serial sections of cell nuclei. Biology of the Cell 62:111-117.
Ghosh S.K. (1981) Quantitative methods applied to stereo imaging. Theory. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:155-177.
Hama K., Arii T. (1987) Three‐dimensional analysis of high‐voltage electron microscope tilt images: methods and problems. J. Electron. Microsc. Technique 6:185-192.
Harauz G., Borland L., Bahr G.F., Zeitler E., Van Heel M. (1987) Three‐dimensional reconstruction of a human metaphase chromosome from electron micrographs. Chromosoma 95:366-374.
Hegerl R. (1989) 3‐D reconstructions from projections in E.M. Eur. J. Cell Biol. 48:135-138.
Hendricks G.M., Sharp W.P. (1987) Quantitative and densitometric considerations in bas‐relief electron micrographs. J. Electron Microsc. Technique 5:203-209.
Hozak P., Novak J.T., Smetana K. (1989) Three‐dimensional reconstruction of nucleolus‐organizing regions in PHA‐stimulated human lymphocytes. Biology of the Cell 66:225-233.
Huijsmans D.P. (1989) The integration of software for computer‐aided 3D‐reconstruction from parallel serial sections; a plea for standards. Eur. J. Cell Biol. 48:57-60.
King M.V. (1981) Theory of stereopsis. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:13-32.
Kristen U., Lockhausen J. (1989) Computer‐generated three‐dimensional representation of dictyosomes reconstructed from electron micrographs of serial ultrathin sections. J. Cell Sci. 93:385-389.
Lindley V.A., Randall W.C., Stumpf D.K. (1986) Bas‐relief for enhancement of photomicrographs. Journal of Electron Microscopy Technique 3:369-370.
Marko M., Leith A., Parsons D. (1988) Three‐dimensional reconstruction of cells from serial sections and whole‐cell mounts using multilevel contouring of stereo micrographs. J. Electron Microsc. Technique 9:395-411.
Moss V.A. (1989) The computation of 3‐D morphology from serial sections. Eur. J. Cell Biol. 48:61-64.
Olins A.L., Olins D.E., Levy H.A., Margle S.M., Tinnel E.P., Durfee R.C. (1989) Tomographic reconstruction from energy‐filtered images of thick biological sections. J. Microsc. 154:257-265.
Peachey L.D., Heath J.P. (1989) Reconstruction from stereo and multiple tilt electron microscope images of thick sections of embedded biological specimens using computer graphic methods. J. Microsc. 153:193-204.
Pebusque M.J., Dupuy‐Coin A.M., Cataldo C., Seite R., Bouteille M., Moens P. (1981) Three‐dimensional electron microscopy of the nucleolar organizer regions (NORs) in sympathetic neurons. Biol. Cell. 41:59-62.
Ploton D., Bobichon H., Adnet J.J. (1982) Ultrastructural localization of NOR in nucleoli of human breast cancer tissues using a one‐step Ag‐NOR staining method. Biol. Cell. 43:229-232.
Ploton D., Beorchia A., Menager M., Jeannesson P., Adnet J.J. (1987) The three‐dimensional ultrastructure of interphasic and metaphasic nucleolar argyrophilic components studied with high‐voltage electron microscopy in thick sections. Biol. Cell. 59:113-120.
Ploton D., Thiry M., Menager M., Lepoint A., Adnet J.J., Goessens G. (1987) Behaviour of nucleolus during mitosis. A comparative ultrastructural study of various cancerous cell‐lines using Ag‐NOR staining procedure. Chromosoma 95:95-107.
Porter K.R., Stearns M.E. (1981) Stereo‐microscopy of whole cells. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:53-76.
Ramachandran V.S. (1988) Perception of shape from shading. Nature 331:163-166.
Rambourg A., Clermont Y. (1990) Three‐dimensional electron microscopy: structure of the Golgi apparatus. Eur. J. Cell Biol. 51:189-200.
Skoglund U., Andersson K., Strandberg B., Daneholt B. (1986) Three‐dimensional structure of a specific pre‐messenger RNP particle established by electron microscope tomography. Nature 319:560-564.
Turner J.N. (1981) Introduction to stereo imaging. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:1-12.
Turner J.N. (1981) Stages and stereo‐pair recording. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:33-52.
Turner J.N., Collins D.N. (1990) Three‐dimensional imaging at high resolution: high‐voltage electron microscopy. J. Histo. Tech. 13:93-107.
Turner J.N., Rieder C.L., Collins D.N., Chang B.B. (1989) Optimum specimen positioning in the electron microscope using a double‐tilt stage. J. Electron Microsc. Technique 11:33-40.
Yamada E., Ishikawa H. (1981) Dense tissue and special stains. Methods in Cell Biology , ed. by, J. N. Turner, Academic Press, London; 22:123-146.
Young S.J., Royer S.M., Groves P.M., Kinnamon J.C. (1987) Three‐dimensional reconstructions from serial micrographs using the IBM PC. J. Electron Microsc. Technique 6:207-217.