Abstract
The technique of cryoelectron tomography of frozen-hydrated biological specimens is opening a new window on cellular structure and organization. This imaging method provides full 3D structural information at much higher resolution (typically 5–10 nm) than is attainable by light microscopy, and can be applied to cells and organelles that are maintained in a state that is as close to native as can be achieved currently in electron microscopy. Not only can cryoelectron tomography be used to visualize directly extended cellular structures, such as membranes and cytoskeleton, but it can also provide 3D maps of the location, orientation and, perhaps, the conformation of large macromolecular complexes, the cell’s ‘molecular machinery’. This information complements that coming from single-particle cryoelectron microscopy (Frank et al., 1996, 2006) and X-ray crystallography, about the subnanometer structure of the same molecular assemblies after isolation. As with studies using single-particle cryoelectron microscopy, specimens smaller than 1 µ in size can be prepared for cryoelectron tomography by plunge-freezing (Dubochet et al., 1988). Cells or organelles can be rapidly frozen directly on an electron microscope grid in thin layers of glass-like, amorphous ice, without the formation of ice crystals that would otherwise disrupt fine structure (Kellenberger, 1987). Specimens are imaged directly, without chemical fixation, dehydration or staining with heavy metals.
Keywords
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsPreview
Unable to display preview. Download preview PDF.
References
Al-Amoudi, A., Dubochet, J., Gnägi, H., Lüthi, W. and Studer, D. (2003). An oscillating cryo-knife reduces cutting-induced deformation of vitreous ultrathin sections. J. Microsc. 212:26–33.
Al-Amoudi A., Norlén, L. P. O. and Dubochet J. (2004). Cryo-electron microscopy of vitreous sections of native biological cells and tissues. J. Struct. Biol. 148:131–135.
Al-Amoudi, A., Studer, D. and Dubochet, J. (2005). Cutting artifacts and cutting process in vitreous sections for cryo-electron microscopy. J. Struct. Biol. 150:109–121.
Baumeister, W. (2002). Electron tomography: towards visualizing the molecular organization of the cytoplasm. Curr. Opin. Struct. Biol. 12:679–684.
Baumeister, W. and Steven, A. C. (2000). Macromolecular electron microscopy in the era of structural genomics. Trends Biochem. Sci. 25:624–631.
Beck, M., Förster, F., Ecke, M., Plitzko, J. M., Melchior, F., Gerisch, G., Baumeister, W. and Medalia, O. (2004). Nuclear pore complex structure and dynamics revealed by cryoelectron tomography. Science 306:1387–1390.
Bernhard, W. and Leduc, E. (1967). Ultrathin frozen sections. J. Cell Biol. 34:757–771.
Böhm, J., Frangakis, A. S., Hegerl, R., Nickell, S., Typke, D. and Baumeister, W. (2000). Toward detecting and identifying macromolecules in a cellular context: template matching applied to electron tomograms. Proc. Natl Acad. Sci. USA 97:14245–14250.
Böhm, J., Lambert, O., Frangakis, A. S., Letellier, L. Baumeister, W. and Rigaud, J. L. (2001). FhuA-mediated phage genome transfer into liposomes: a cryo-electron tomography study. Curr. Biol. 11:1168–1175.
Bongini, L., Fanelli, D., Piazza, F., De los Rios, P., Sandin, S. and Skoglund, U. (2004). Freezing immunoglobulins to see them move. Proc. Natl Acad. Sci. USA 101:6466–6471.
Brandt, S., Heikkonen, J. and Engelhardt, P. (2001a). Multiphase method for automatic alignment of transmission electron microscope images using markers. J. Struct. Biol. 133:10–22.
Brandt, S., Heikkonen, J. and Engelhardt, P. (2001b). Automatic alignment of transmission electron microscope tilt series without fiducial markers. J. Struct. Biol. 136:201–213.
Bretschneider. T., Jonkman, J., Kohler, J., Medalia, O., Barisic, K. Weber, I., Selzer, E. H. K., Baummeister, W. and Gerisch, G. (2002). Dynamic organization of the actin system in the motile cells of Dictyostelium. J. Muscle Res. Cell Motil. 23:639–649.
Buchanan, R. A., Leapman, R. D., O’Connell, M. F., Reese, T. S. and Andrews, S. B. (1993). Quantitative scanning transmission electron microscopy of ultrathin cryosections: subcellular organelles in rapidly frozen liver and cerebellar cortex. J. Struct. Biol. 110:244–255.
Chang, J.-J., McDowall, A.W., Lepault, J., Freeman, R., Walter, C.A. and Dubochet, J. (1983). Freezing, sectioning and observation artifacts of frozen hydrated sections for electron microscopy. J. Microsc. 132:109–123.
Craig, S. and Staehelin, L. A. (1988). High pressure freezing of intact plant tissues. Evaluation and characterization of novel features of the endoplasmic reticulum and associated membrane systems. Eur. J. Cell Biol. 46:81–93.
Crowther, R. A., DeRosier, D. J. and Klug, A. (1970). The reconstruction of a three-dimensional structure from its projections and its applications to electron microscopy. Proc. R. Soc. B 317:319–340.
Cyrklaff, M., Risco, C., Fernandez, J. J., Jimenez, M. V., Esteban, M., Baumeister, W. and Carrascosa, J. L. (2005). Cryo-electron tomography of Vaccinia virus. Proc. Natl Acad. Sci. USA 102:2772–2777.
Dahl, R. and Staehelin, L. A. (1989). High-pressure freezing for the preservation of biological structure: theory and practice. J. Electron Microsc. Tech. 13:165–174.
Dierksen, K., Typke, D., Hegerl, R. and Baumeister, W. (1993). Towards automatic electron tomography. II. Implementation of autofocus and low-dose procedures. Ultramicroscopy 49:109–120.
Dierksen, K., Typke, D., Hegerl, R., Walz, J., Sackmann, E. and Baumeister, W. (1995). Three-dimensional structure of lipid vesicles embedded in vitreous ice and investigated by automated electron tomography. Biophys. J. 68:1416–1422.
Dubochet, J., Adrian, M., Chang, J. J., Homo, J.-C., Lepault, J., McDowall, A.W. and Schultz, P. (1988). Cryo-electron microscopy of vitrified specimens. Q. Rev. Biophys. 21:129–228.
Dubochet, J., Adrian, M., Chang, J.-J., Lepault, J. and McDowall, A. (1987). Cryoelectron microscopy of vitrified specimens. In Cryotechniques in Biological Electron Microscopy (R. A. Steinbrecht and K. Zierold, eds). Springer, Berlin, pp. 114–131.
Dubochet, J. and Sartori Blanc, N. (2001). The cell in absence of aggregation artifacts. Micron 32:91–99.
Echlin, P. (1992). Low-temperature Microscopy and Microanalysis. Plenum, New York.
Edelman, L. (1994). Optimal freeze-drying of cryosections and bulk specimens for X-ray microanalysis. Scanning Microsc. Suppl. 8:67–81.
Erk, I., Nicolas, G., Carloff, A. and Lepault, J. (1998). Electron microscopy of frozen biological objetcs: a study using cryosectioning and cryosubstitution. J. Microsc. 189:236–248.
Fernández-Morán, H. (1952). Application of the ultrathin freeze sectioning technique to the study of cell structures with the electron microscope. Arch. Fysik. 4:471–483.
Fernández-Morán, H. (1960). Low-temperature preparation techniques for electron microscopy of biological specimens based on rapid freezing with liquid helium II. Annu. NY Acad. Sci. 85:689–713.
Forbes, M. S. (1986). Dog hairs as section manipulators. EMSA Bull. 16:67.
Förster, F., Medalia, O., Zauberman, N., Baumeister, W. and Fass, D. (2005). Retrovirus envelope protein complex structure in situ studied by cryo-electron tomography. Proc. Natl Acad. Sci. USA 102:4729–4734.
Frangakis, A. S., Böhm, J., Förster, F., Nickell, S., Nicastro, D., Typke, D., Hegerl, R. and Baumeister, W. (2002). Identification of macromolecular complexes in cryoelectron tomograms of phantom cells. Proc. Natl Acad. Sci. USA 99:14153–14158.
Frangakis, A. S. and Förster, F. (2004). Computational exploration of structural information from cryo-electron tomograms. Curr. Opin. Struct. Biol. 14:325–331.
Frank, J. (1996). Three-dimensional Electron Microscopy of Macromolecules. Academic Press, San Diego.
Frank, J. (2006). Three-dimensional Electron Microscopy of Macromolecular Assemblies—Visualization of Biological Molecules in Their Native State. Oxford University Press, New York.
Frank, J., Wagenknecht, T., McEwen, B. F., Marko, M., Hsieh, C.-E. and Mannella, C. A. (2002). Three-dimensional imaging of biological complexity. J. Struct. Biol. 138:85–91.
Frederik, P. M., Bomans, P. H. H. and Stuart, M. C. A. (1993). Matrix effects and the induction of mass loss or bubbling by the electron beam in vitrified hydrated specimens. Ultramicroscopy 48:107–119.
Frederik, P. M., Busing, W. M. and Persson, A. (1982). Concerning the nature of the cryosectioning process. J. Microsc. 125:167–175.
Frederik, P. M., Busing, W. M. and Persson, A. (1984). Surface defects on thin cryosections. Scanning Electron Microsc. 1:433–443.
Gessler, A. E. and Fullam, E. F. (1946). Sectioning for the electron microscope accomplished by the high speed microtome. Am. J. Anat. 78:245–279.
Gilkey, J. C. and Staehelin, L. A. (1986). Advances in ultrarapid freezing for the preservation of cellular ultrastructure. J. Electron Microsc. Tech. 3:177–210.
Grimm, R., Singh, H., Rachel, R., Typke, D., Zilling, W. and Baumeister, W. (1998). Electron tomography of ice-embedded prokaryotic cells. Biophys. J. 74:1031–1042.
Grünewald, K., Desai, P., Winkler, D. C., Heyman, J. B., Belnap, D. M., Baumeister, W. and Steven, A. C. (2003). Three-dimensional structure of herpes simplex virus from cryoelectron tomography. Science 302:1396–1398.
Grünewald, K., Medallia, O., Gross, A., Steven, A. and Baumeister, W. (2002). Prospects of electron cryotomography to visualize macromolecular complexes inside cellular compartments: implications of crowding. Biophys. Chem. 100:577–591.
Gupta, B. L. and Hall, T.A. (1981). The x-ray microanalysis of frozen-hydrated sections in scanning electron microscopy: an evaluation. Tissue and Cell 13:623–643.
Hama, K. and Arii, T. (1987). Three-dimensional analysis of high-voltage electron microscope tilt images: methods and problems. J. Electron Microsc. Tech. 6:185–192.
Hess, M.W., Muller, M., Debbage, P. L., Vetterlein, M. and Pavelka, M. (2000). Cryopreparation provides new insight into the effects of brefeldin A on the structure of the HepG2 Golgi apparatus. J. Struct. Biol. 130:63–72.
Hodson, S. and Marshall, J. (1970). Ultracryotomy: a technique for cutting ultrathin sections of unfixed biological tissues for electron micrscopy. J. Microsc. 91:105–117.
Hohenberg, H., Tobler, M. and Müller, M. (1996). High-pressure freezing of tissue obtained by fine-needle biopsy. J. Microsc. 183:1–7.
Hsieh, C., He, W., Marko, M. and Stokes, D. L. (2004). 3D Tomographic map of desmosome from frozen-hydrated skin sections. Microsc. Microanal. 10(Suppl. 2):1188CD.
Hsieh, C.-E., Leith, A., Mannella, C.A., Frank, J. and Marko, M. (2006). Towards high-resolution three-dimensional imaging of native mammalian tissue: electron tomography of frozen-hydrated rat liver sections. J. Struct. Biol. 153 (in press).
Hsieh, C.-E., Marko, M., Frank, J. and Mannella, C. A. (2002). Electron tomographic analysis of frozen-hydrated tissue sections. J. Struct. Biol. 138:63–73.
Hsieh, C.-E., Marko, M., Leith, A., Frank, J. and Mannella, C. A. (2003). Electron tomographic comparison of frozen-hydrated and freeze-substituted sections of high-pressure frozen rat-liver tissue. Microsc. Microanal. 9(Suppl. 2):1178CD.
Hutchinson, T. E., Johnson, D. E. and MacKenzie, A.P. (1978). Instrumentation for direct observation of frozen hydrated specimens in the electron microscope. Ultramicroscopy 3:315–324.
Kellenberger, E. (1987). The response of biological macromolecules and supremolecular structures to the physics of specimen cryopreparation. In Cryotechniques in Biological Electron Microscopy (R. A. Steinbrecht and K. Zierold, eds). Springer, Berlin, pp. 35–63.
Koster, A. J., Grimm, R., Typke, D., Hegerl, R., Stoschek, A., Walz, J. and Baumeister, W. (1997). Perspectives of molecular and cellular electron tomography. J. Struct. Biol. 120:276–308.
Kürner, J., Medalia, O., Linaroudis, A. A. and Baumeister, W. (2004). New insights into the structural organization of eukaryotic and prokaryotic cytoskeletons using cryo-electron tomography. Exp. Cell Res. 301:38–42.
Kürner, J., Frangakis, A. S. and Baumeister, W. (2005). Cryo-electron tomography reveals the cytoskeletal structure of Spiroplasma melliferum. Science 307:436–438.
Leapman, R. D. (2005). Novel techniques in electron microscopy. Curr. Opin. Neurobiol. 14:591–598.
Leforestier, A., Dubochet, J. and Livolant. F. (2001). Bilayers of nucleosome core particles. Biophys. J. 81:2414–2421.
Leis, A., Andrees, L., Gruska, M., Al-Amoudi, A., Sartori, A., Dubochet, J. and Baumeister, W. (2005). Cryo-electron tomography and fluorescence microscopy of unicellular algae in vitreous cryosections. Microsc. Microanal. 11(Suppl. 2):330CD.
Lepault, J., Bigot, D., Studer, D. and Erk, I. (1997). Freezing of aqueous specimens: an X-ray diffraction study. J. Microsc. 187:158–166.
Lucic, V., Förster, F. and Baumeister, W. (2005b). Structural studies by electron tomography: from cells to molecules. Annu. Rev. Biochem. 74:833–865.
Lucic, V., Yang, T., Schweikert, G., Förster, F. and Baumeister, W. (2005a). Morphological characterization of molecular complexes present in the synaptic cleft. Structure 13:423–434.
Luther, P. K., Lawrence, M. C. and Crowther, R.A. (1988) A method for monitoring the collapse of plastic sections as a function of electron dose. Ultramicroscopy 24:7–18.
Mannella, C. A. (2006). The relevance of mitochondrial membrane topology to mitochondrial function. Biochem. Biophys. Acta 1762:140–147.
Mannella, C. A., Buttle, K., Marko, M. (1997). Reconsidering mitochondrial structure: new views of an old organelle. Trends Biochem. Sci. 22:37–38.
Mannella, C. A. and Frey, T. (2000). The internal structure of mitochondria. Trends Biochem. Sci. 25:319–324.
Mannella, C. A., Pfeiffer, D. R., Bradshaw, P. C., Moraru, L. I., Slepchenko, L. B., Loew, L. M., Hsieh, C.-E., Buttle, K. and Marko, M. (2001). Topology of the mitochondrial inner membrane: dynamics and bioenergetic implications. IUBMB Life 52:93–100.
Marko, M., Hsieh, C.-E., Mannella, C. A. and Frank, J. (2002). Electron tomography of frozen-hydrated specimens: application to tissue sections. In Proceedings of the 15th International Congress on Electron Microscopy (Cross, R., ed.), Microscopy Society of Southern Africa, Onderspoort, SA, 2:205–206.
Marko, M., Hsieh, C.-E., Mannella, C. A. and McEwen, B. (1999). Imaging considerations for cryo-tomography of organelles and whole cells at high accelerating voltage. Microsc. Microanal. 5(Suppl. 2):414–415.
Marko, M., Hsieh, C., MoberlyChan, W., Mannella, C.A. and Frank, J. (2006). Focused ion beam milling of vitreous water: prospects for an alternative to cryo-ultramicrotomy. J. Microsc. 212:42–47.
Marko, M., Hsieh, C.-E., Rath, B. K., Mannella, C. A. and McEwen, B. F. (2000). Electron tomography of frozen-hydrated samples. Microsc. Microanal. 6(Suppl. 2):310–311.
Mastronarde, D. N. (1997). Dual-axis tomography: an approach with alignment methods that preserve resolution. J. Struct. Biol. 120:343–352.
Mastronarde, D. N. (2005). Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152:36–51.
Matias, V. R. F., Al-Amoudi, A., Dubochet, J. and Beveridge, T. J. (2003). Cryo-transmission electron microscopy of frozen-hydrated sections of gram-negative bacteria. J. Bacteriol. 185:6112–6118.
McDonald, K. and Morphew, M. K. (1993). Improved preservation of ultrastructure in difficult-to-fix organisms by high pressure freezing and freeze substitution: 1 Drosophila melanogaster and Strongylocentrotus purpuratus embryos. Microsc. Res. Tech. 24:254–473.
McDowall, A.W., Chang, J. J., Freeman, R., Lepault, J., Walter, C.A. and Dubochet, J. (1983). Electron microscopy of frozen hydrated sections of vitreous ice and vitrified biological samples. J. Microsc. 131:1–9.
McEwen, B. F. and Frank, J. (2001). Electron tomographic and other approaches for imaging molecular machines. Curr. Opin. Neurobiol. 11:594–600.
McEwen, B. F., Marko, M., Hsieh, C.-E. and Mannella, C. (2002). Use of frozen-hydrated axonemes to assess imaging parameters and resolution limits in cryoelectron tomography. J. Struct. Biol. 138:47–57.
McIntosh, J. R. (2001) Electron microscopy of cells: a new beginning for a new century. J. Cell Biol. 153:F25–F32.
McIntosh, J. R., Nicastro, D. and Mastronarde, D. (2005). New views of cells in 3D: an introduction to electron tomography. Trends Cell Biol. 15:43–51.
Medalia, O., Weber, I., Frangakis, A. S., Nicastro, D., Gerisch, G. and Baumeister, W. (2002). Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 289:1209–1213.
Michel, M., Hillman, T. and Müller, M. (1991). Cryosectioning of plant material frozen at high pressure. J. Microsc. 163:3–18.
Michel, M., Gnägi, H. and Müller, M. (1992). Diamonds are a cryosectioner’s best friend. J. Microsc. 166:43–56.
Moor, H. (1987). Theory and practice of high pressure freezing. In Cryotechniques in Biological Electron Microscopy (R. A. Steinbrecht and K. Zierold, eds). Springer, Berlin, pp. 175–191.
Morphew, M. K. and McIntosh, J. R. (2003). The use of filter membranes for high-pressure freezing of cell monolayers. J. Microsc. 212:21–25.
Nicastro, D., Austin, J., Pierson, J., Gaudette, R., Schwartz, C., Ladinsky, M., Staehelin, L. A. and McIntosh, J. R. (2005b). Visualizing the macromolecular organization of chloroplast membranes using cryo-electron tomography. Microsc. Microanal. 11(Suppl. 2):150–151.
Nicastro, D., Frangakis, A. S., Typke, D. and Baumeister, W. (2000). Cryoelectron tomography of Neurospora mitochondria. J. Struct. Biol. 129:48–56.
Nicastro, D., McIntosh, J. R. and Baumeister, W. (2005a). 3-D structure of eukaryotic flagella in a quiescent state revealed by cryo-electron tomography. Proc. Natl Acad. Sci. USA 102:15889–15894.
Nickell, S., Hegerl, R., Baumeister, W. and Rachel, R. (2003). Pyrodictium cannulae enter the periplasmic space but do not enter the cytoplasm, as revealed by cryo-electron tomography. J. Struct. Biol. 141(1):34–42.
Norlén, L. and Al-Amoudi, A. (2004). Stratum corneum keratin structure, function and formation—the cubic rod-packing and membrane templating model. J. Invest. Dermatol. 123:715–732.
Norlén, L., Al-Amoudi, A. and Dubochet, J. (2003). A cryotransmission electron microscopy study of skin barrier formation. J. Invest. Dermatol. 120:555–560.
Penczek, P., Marko, M., Buttle, K. and Frank, J. (1995). Double-tilt electron tomography. Ultramicroscopy 60:393–410.
Plitzko, J. M., Frangakis, A. S., Foerster, F., Gross, A. and Baumeister, W. (2002). In vivo veritas: electron cryotomography of intact cells with molecular resolution. Trends Biotechnol. 20:40–44.
Rath, B. K., Hegerl, R., Leith, A., Shaikh, T. R., Wagenknecht, T. and Frank, J. (2003). Fast 3D motif search of EM density maps using a locally normalized cross-correlation function. J. Struct. Biol. 144:95–103.
Rath, B. K., Marko, M., Radermacher, M. and Frank, J. (1997). Low-dose automated electron tomography: a recent implementation. J. Struct. Biol. 120:210–218.
Richter, K. (1994). Cutting artefacts on ultrathin cryosections of biological bulk specimens. Micron 25:297–308.
Richter, K. (1996). Aspects of cryofixation and cryosectioning for the observation of bulk biological samples in the hydrated state by cryoelectron microscopy. Scanning Microsc. (Suppl. 10):375–386.
Richter, K., Gnägi, H. and Dubochet, J. (1991). A model for cryosectioning based on the morphology of vitrified ultrathin sections. J. Microsc. 163:19–28.
Rockel B., Jakana J., Chiu W. and Baumeister W. (2002). Electron cryo-microscopy of VAT, the archaeal p97/CDC48 homologue from Thermoplasma acidophilum. J. Mol. Biol. 317:673–668.
Sali, A., Glaeser, R., Earnest, T. and Baumeister, W. (2003). From words to literature in structural proteomics. Nature 422:216–225.
Sandin S., Ofverstedt L.G., Wikstrom, A.C., Wrange, O. and Skoglund, U. (2004). Structure and flexibility of individual immunoglobulin G molecules in solution. Structure 12:409–415.
Sartori, N., Bednar, J. and Dubochet, J. (1996). Electron-beam-induced amorphization of ice III or IX obtained by high-pressure freezing. J. Microsc. 182:163–168.
Sartori, N., Richter, K. and Dubochet, J. (1993). Vitrification depth can be increased more than 10-fold by high-pressure freezing. J. Microsc. 172:55–61.
Sartori Blanc, N., Studer, D., Ruhl, K. and Dubochet, J. (1998). Electron beam-induced changes in vitreous sections of biological samples. J. Microsc. 192:194–201.
Sartori-Blanc, N., Senn, A., Leforestier, A., Livolant, F. and Dubochet, J. (2001) DNA in human and stallion spermatozoa forms local hexagonal packing with twist and many defects. J. Struct. Biol. 134:76–81.
Sawaguchi, A., Yao, X., Forte, J. G. and McDonald, K. L. (2003). Direct attachment of cell suspensions to high-pressure freezing specimen planchettes. J. Microsc. 212:13–20.
Schwartz, C., Nicastro, D., Ladinsky, M. S., Mastronarde, D., O’Toole E. and McIntosh, J. R. (2003). Cryo-electron tomography of frozen-hydrated sections of eukaryotic cells. Microsc. Microanal. 9(Suppl. 2):1166–1167.
Shi, S., Sun, S.Q., Andrews, B. and Leapman, R.D. (1996). Thickness measurement of hydrated and dehydrated cryosections by EELS. Microsc. Res. Tech. 33:241–250.
Shimoni, E. and Müller, M. (1998). On optimizing high-pressure freezing: from heat transfer theory to a new microbiopsy device. J. Microsc. 192:236–247.
Sitte, H. (1996). Advanced instrumentation and methodology related to cryoultramicrotomy: a review. Scanning Microsc. (Suppl. 10):387–466.
Somlyo, A. V., Shuman, H. and Somlyo, A. P. (1977). Elemental distribution of striated mucle and the effects of hypertonicity. J. Cell Biol. 74:824–857.
Somlyo, A. P., Bond, M. and Somlyo, A.V. (1985). Calcium content of mitochondria and endoplasmic reticulum in liver frozen rapidly in vivo. Nature 314:622–625.
Steinbrecht, R. A. and Zierold, K. (1987). Cryotechniques in Biological Electron Microscopy. Springer, Berlin.
Steven A. C. and Aebi, U. (2003). The next ice age: cryo-electron tomography of intact cells. Trends Cell Biol. 13:107–110.
Stoffler, D., Feja, B., Fahrenkrog, J., Walz, J., Typke, D. and Aebi, U. (2003). Cryo-electron tomography provides novel insights into nuclear pore architecture: implications for nucleocytoplasmic transport. J. Mol. Biol. 328:119–130.
Studer, D. and Gnägi, H. (2000). Minimal compression of ultrathin sections with use of an oscillating diamond knife. J. Microsc. 197:94–100.
Studer, D., Graber, W., Al-Amoudi, A. and Eggli, P. (2001). A new approach for cryofixation by high-pressure freezing. J. Microsc. 203:285–294.
Studer, D., Michel, M. and Müller, M. (1989). High pressure freezing comes of age. Scanning Microsc. (Suppl. 3):253–269.
Studer, D., Michel, M., Wohlwend, M., Hunziker, E. B. and Buschmann, M. D. (1995). Vitrification of articular cartilage by high-pressure freezing. J. Microsc. 179 321–332.
Sun, S.Q., Shi, S.-L., Hunt, J.A., Leapman, R.D. (1995). Quantitative water mapping of cryosectioned cells by electron energy-loss spectroscopy. J. Microsc. 177:18–30.
Taylor, K.A. and Glaeser, R.M. (1974). Electron diffraction of frozen, hydrated protein crystals. Science 186:1036–1037.
Taylor, K. A. and Glaeser, R. M. (1976). Electron microscopy of frozen hydrated biological specimens. J. Ultrastruct. Res. 55:448–456.
Ting, C. S., Hsieh, C., Sundararaman, S., Mannella, C. A. and Marko, M. (2005). Comparative three-dimensional imaging of environmentally critical cyanobacteria through cryoelectron tomography. Microsc. Microanal. 11(Suppl. 2): 332CD.
Tokuyasu, K. T. (1986). Application of cryoultramicrotomy to immunocytochemistry. J. Microsc. 143:139–149.
Vanhecke, D., Graber, W., Herrmann, G., Al-Amoudi, A., Eggli, P. and Studer. D. (2003). A rapid microbiopsy system to improve the preservation of biological samples prior to high-pressure freezing. J. Microsc. 212:3–12.
Van Marle, J., Dietrich, A., Jonges, K., Jonges, R, de Moor, E., Vink, A., Boon, P. and van Veen, H. (1995). EM-tomography of section collapse, a non-linear phenomenon. Microsc. Res. Tech. 31:311–316.
Vonk, J. (2000). Parameters affecting specimen flatness of two-dimensional crystals for electron crystallography. Ultramicroscopy 5:123–129.
Wagenknecht, T., Hsieh, C.-E., Rath, B., Fleischer, S. and Marko, M. (2002). Electron tomography of frozen-hydrated isolated triad junctions. Biophys. J. 83:2491–2501.
Walther, P. and Müller, M. (1997). Double-layer coating for field-emission cryo-scanning electron microscopy—present state and applications. Scanning 19:343–348.
Walz, J., Typke, D., Nitsch, M., Koster, A. J., Hegerl, R. and Baumeister, W. (1997). Electron tomography of single ice-embedded macromolecules: three-dimensional alignment and classification. J. Struct. Biol. 120:387–395.
Winkler, H. and Taylor, K.A. (2006). Accurate marker-free alignment with simultaneous geometry determination and reconstruction of tilt series in electron tomography. Ultramicroscopy 106:240–254.
Woodcock, C. L. (1994). Chromatin fibers observed in situ in frozen hydrated sections. Native fiber diameter is not correlated with nucleosome repeat length. J. Cell Biol. 125:11–19.
Zhang, P., Bos, E., Heyman, J., Gnägi, H., Kessel, M., Petere, P. J. and Subramaniam, S. (2004). Direct visualization of receptor arrays in frozen-hydrated sections and plunge-frozen specimens of E. coli engineered to overproduce the chemotaxis receptor Tsr. J. Microsc. 216:76–83.
Zhao, Q., Ofverstedt, L. G., Skoglund, U. and Isaksson, L. A. (2004). Morphological variation of individual Escherichia coli 30S ribosomal subunits in vitro and in situ, as revealed by cryo-electron tomography. Exp. Cell Res. 297:495–507.
Zierold, K. (1984). The morphology of ultrathin cryosections. Ultramicroscopy 14:201–210.
Zierold, K. (1987). Cryoultramicrotomy. In Cryotechniques in Biological Electron Microscopy (R. A. Steinbrecht and K. Zierold, eds). Springer, Berlin, pp. 132–148.
Zierold, K. (1988). X-ray microanalysis of freeze-dried and frozen-hydrated cryosections. J. Electron Microsc. Tech. 9:65–82.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Marko, M., Hsieh, CE., Mannella, C.A. (2007). Electron Tomography of Frozen-hydrated Sections of Cells and Tissues. In: Frank, J. (eds) Electron Tomography. Springer, New York, NY. https://doi.org/10.1007/978-0-387-69008-7_3
Download citation
DOI: https://doi.org/10.1007/978-0-387-69008-7_3
Publisher Name: Springer, New York, NY
Print ISBN: 978-0-387-31234-7
Online ISBN: 978-0-387-69008-7
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)