Abstract
Traditionally, biologists have been confined to electron microscopy and light microscopy in order to correlate biochemical and molecular data with morphology. Electron microscopy provides fine ultrastructural detail, but tends to limit one to the study of cellular structures that react with electron-dense stains deposited in fixed specimens. Immunogold labeling allows one to study non electron-dense material, but the EM sections must be relatively thin, and even then there are problems with penetration of the labelled antibodies. Another problem is the necessity of making thin sections to prevent scattering of the electron beam. The electron microscopist is forced to live like a citizen of Flatland. Flatland is a story about beings that existed entirely within a plane, and were trying to figure out what the various two dimensional shapes appearing on their “world” were (Abbott, 1884). The shapes were sections of three dimensional objects passing through their plane of being. Similarly, biologists using an electron micrsoscope attempt to reconstruct the three dimensional structure of a cell based on the two dimensional cuts through the specimen.
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References
Abbott, E. (1884) Fiatland: A Romance of Many Dimensions, 2nd ed, Dover, London.
Bacallao, R., Dotti, G, Antony, G, Stelzer, E.H.K., Karsenti, E and K. Simons, (1989) Subcellular organization of MDCK cells during the formation of a polarized epithelium, J. Cell Biol. (109, 2817–32).
Bacallao, R., Jesaitis, L., Karsenti, E and J. De Mey, (1990) The Effects of Fixation On Shrinkage-Induced Artifacts. Submitted, J. Histochem & Cytochem.
Bacallao, R., Steltzer, E. H. K., (1989) Preservation of BIOLOGICAL Specimens for Observation in a Confocal Fluorescence Microscope, Operational Principles of Confocal Fluorescence Microscopy Methods in Cell Biology, Vol. 31, 437–452
Bastholm, L., Scopsi, L., and Nielsen, M.H. (1986) Silver-enhanced immunogold staining of semithin and ultrathin cryosections, J. Electron Microsc. Technique, 4, 175–6.
Berod, A., Hartman, B.K. and Pujol, J.F. (1981) Importance of fixation in immunohistochemistry, J. of Histochem and Cytochem. 29: 844–50.
Birrell, G.B. and K.K. Hedbert (1987) Immunogold labeling with small gold particles: Siver enhancement provides increased detectability at low magnifications, J. Electron Microsc. Technique, 5, 219–20.
Bock, G., Hilchenbach, M., Schauenstein, K., and G Wick (1985) Photometric analysis of anti-fading reagents for immunofluorescence with laser and conventional illumination sources, J. Histochem. Cytochem., 33, 699–705.
Bomsel, M., Prydz, K., Parton, R.G, Gruenberg, J., and K. Simons, Functional and topological organization of apical and basolateral endocytic pathways in MDCK cells, manuscript submitted.
Cande, W.Z., Lazarides, E., and J.R. Mcintosh (1977) Composition and distribution of actin and tubulin in mammalian mitotic spindle as seen by indirect immunofluorescence, J. Cell. Biol., 72, 552–567.
Dabora, S.L. and M.P. Sheetz (1988) The microtubule-dependent formation of tubulovesicular network with characteristics of the endoplasmic-reticulum from cultured cell extracts, Cell, 54, 27–35.
Danscher, G., Rytter Nørgaard, J.O., and E. Baatrup (1987) Autome-tallography-tissue metals demonstrated by a silver enhancement kit, Histochemistry, 71, 1–16.
Fox, G.H., Johnson, F.B., Whiting, J., and P.P. Roller (1985) Formaldehyde fixation, J. Histochem. Cytochem., 33, 845–53.
Holgate, G.S., Jackson, P., Cowen, P.N., and GG Bird (1983) Immu-nogold-silver staining: New method of immunostaining with enhanced sensitivity, J. Histochem. Cytochem., 31, 938–44.
Inoué, S., (1986) Video Microscopy, New York and London: Plenum Press
Ito, S., (1962) Light and electron microscopic study of membranous cytoplasmic organelles, In The Interpretation of Ultrastructure, R.J.C. Harris, ed. New York: Academic Press, pp. 129–148.
Lackie, P.M., Hennessy, R.J., Hacker, G.W., and J.M. Polak (1985) Investigation of immunogold-silver staining by electron microscopy, Histochemistry, 83, 545–50.
Langanger, G., De Mey, J., and H. Adam (1983) 1,4-Diazobizyklo-[2.2.2]oktan (DABCO) verzogest das Ausbleichen von immunoflu-oreszenzpraparaten, Mikroskopie, 40, 237–41.
Lee, G., and L.B. Chen (1988) Dynamic behavior of endoplasmic-reticulum in living cells, Cell, 54, 37–46.
Lee, R.M.K. (1984) A critical appraisal of the effects of fixation, dehydration and embedding on cell volume, in The Science of Biological Specimen Presevationfor Microscopy and Microanalysis., ed Revel, J-R, Barnard, T., Haggis, GH., and S.A. Bhatt, SEM Inc., AMF O’Hare, Chicago, Ill., 61–70.
Lee, R.M.K., Garfield, R.E., Forrest, J.B., and E.E. Daniel (1979) The effects of fixation, dehydration and critical point drying on the size of cultured smooth-muscle cells, Scanning Electron Microsc. 179, III, 439–448.
Lee, R.M.K., Garfield, R.E., Forrest, J.B., and E.E. Daniel (1980) Dimensional changes of cultured smooth muscle cells due to preparatory processes for transmission electron-microscopy, J. Microsc, 120, 85–91.
Lee, R.M.K., McKenzie, R., Kobayashi, K., Garfield, R.E, Forrest, J.B., and E.E. Daniel (1982) Effects of glutaraldehyde fixative osmolalities on smooth-muscle cell-volume and osmotic reactivity of the cells after fixation, J. Microsc, 125, 77–88.
Lipsky, N. and Pagano, R.E. (1985) A vital stain for the Golgi apparatus, J. Cell Biol. 100: 27–34.
McLean, I.W. and P.K. Nakane (1974) Periodate-lysine-formaldehyde fixative-a new fixative for immunoelectron microscopy, J. Histochem.Cytochem. 22, 1077–1083.
Merdes, A., and J. De Mey, manuscript in preparation.
Nakane, P. (1975) Recent progress in peroxidase-labeled antibody method, Ann. N.Y. Acad. Sci., 254, 203–210.
Osborn, M. (1981) in Techniques in Cellular Physiology part 1, ed. P.F. Baker, Elsevier North Holland Inc., New York, N.Y., 107/1–28.
Paddock, S.W., Tandem scanning reflected light microscopy of cell substratum adhesions and stress fibers in Swiss 3T3 cells, J. Cell Science, 93: 143–146.
Rambourg, A., Clermont, Y., Hermo, L., and D. Segretain (1987) Tridimensional structure of the Golgi apparatus of non-ciliated epithelial cells of the ductuli efferentes in rat—an electron microscope stereoscopic study, Biology of the Cell, 60, 103–116.
Robertson, J.D., Bodenheimer, T.S., and D.E. Stage (1963) Ultrastructure of Mauthner cell synapses and nodes in goldfish brains, J. Cell Biol., 19, 159–199.
Sato, H., Ohnuki, Y, and K. Fujiwara (1976) Immunoflourescent anti-tubulin staining of spindle microtubules and critique for the technique, in Cell Motility. R. Goldman, T. Pollard, and J. Rosenbaum, editors. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y, 419–33.
Scopsi, L., and L.-I. Larsson (1985) Increased sensitivity in immuno-cytochemistry-effects of double amplification of antibodies and of silver intensification on immunogold and peroxidase-antiperoxidase staining techniques, Histochemistry, 82, 321–29.
Sheetz, M.P. and Spudich, J.A. (1983) Movement of myosin-coated fluorescent beads on actin cables in vitro, Nature, 303, 31–35.
Terasaki, M., Song, J., Wong, J.R., Weiss, M.J., and L.B. Chen (1984) Localization of endoplasmic-reticulum in living and glutaraldehyde-fixed cells with fluorescent dyes, Cell, 38, 101–108.
Tooze, J. (1964) Measurements of some cellular changes during fixation of amphibian erythrocytes with osmium tetroxide solutions, J. Cell Biol., 22, 551–63.
van Meer, G., Stelzer, E.H.K., Wijnaendts van Resandt, R.W., Simons, K. (1987) Sorting of glycolipids in epithelial (Madin-Darby canine kidney) cells, J. Cell Biol., 105: 1623–35.
Walsh, M.L., Jen, J., and L.B. Chen (1979) Transport of serum components into structures similar to mitochondria, Cold Spring Harbor Conf. Cell Prolif., 6, 513–20.
Wangesteen, D., Bachofen, H., and E.R. Weibel, (1981) Effects of glutaraldehyde or osmium tetroxide fixation on the osmotic properties of lung cells, J. Microsc, 124, 189–96.
Weber, K., Rathke, P.C., and M. Osborn (1978) Cytoplasmic microtubular images in glutaraldehyde-fixed tissure culture cells by electron microscopy and by immunofluorescence microscopy, Proc. Natl. Acad. Sci. USA., 75, 1820–24.
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© 1990 Plenum Press, New York
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Bacallao, R., Bomsel, M., Stelzer, E.H.K., De Mey, J. (1990). Guiding Principles of Specimen Preservation for Confocal Fluorescence Microscopy. In: Pawley, J.B. (eds) Handbook of Biological Confocal Microscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-7133-9_18
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DOI: https://doi.org/10.1007/978-1-4615-7133-9_18
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