High-resolution electron microscopy of biological specimens in cubic ice.

Images of two biological test specimens, catalase and TMV, were recorded in cubic ice, prepared by controlled devitrification at -130 degrees C and -75 degrees C or in vitrified buffer. Cubic ice provides a rigid support for biological specimens which is stable under the electron beam to within about 1 A, as shown by images of the ice lattice. Neither catalase nor TMV were disrupted by the crystallization of vitrified water. Electron diffraction patterns of highly oriented rafts of TMV extending to 2.3 A resolution were used to judge the quality of TMV images. Structural high-resolution details of TMV was better in cubic ice, and the success rate for recording good images was higher than in vitrified medium.

[1]  Yoshinori Fujiyoshi,et al.  Atomic model of plant light-harvesting complex by electron crystallography , 1994, Nature.

[2]  R. Henderson,et al.  A side-entry cold holder for cryo-electron microscopy. , 1991, Ultramicroscopy.

[3]  N. G. Wrigley,et al.  The lattice spacing of crystalline catalase as an internal standard of length in electron microscopy. , 1968, Journal of ultrastructure research.

[4]  M. Dougherty,et al.  Computer-controlled spot-scan imaging of crotoxin complex crystals with 400 keV electrons at near-atomic resolution. , 1992, Ultramicroscopy.

[5]  K. Gehring,et al.  Structural architecture of an outer membrane channel as determined by electron crystallography , 1991, Nature.

[6]  J. Dubochet,et al.  Cryo-electron microscopy of viruses , 1984, Nature.

[7]  W. Chiu,et al.  Analysis of symmetry and three-dimensional reconstruction of thin gp32*I crystals. , 1991, Journal of molecular biology.

[8]  K. Downing,et al.  Two-dimensional structure of plant light-harvesting complex at 37 Å resolution by electron crystallography☆ , 1989 .

[9]  R. Schröder.  Zero‐loss energy‐filtered imaging of frozen‐hydrated proteins: model calculations and implications for future developments , 1992, Journal of microscopy.

[10]  P. Unwin,et al.  Beef liver catalase structure: interpretation of electron micrographs. , 1975, Journal of molecular biology.

[11]  W. Kühlbrandt,et al.  Structure of light-harvesting chlorophyll a/b protein complex from plant photosynthetic membranes at 7 A resolution in projection. , 1988, Journal of molecular biology.

[12]  G. Büldt,et al.  Densely packed β-structure at the protein-lipid interface of porin is revealed by high-resolution cryo-electron microscopy , 1989 .

[13]  J. Lepault,et al.  Structure of purple membrane from halobacterium halobium: recording, measurement and evaluation of electron micrographs at 3.5 Å resolution , 1986 .

[14]  N. Unwin,et al.  Contrast transfer for frozen-hydrated specimens: determination from pairs of defocused images. , 1988, Ultramicroscopy.

[15]  J. Dubochet,et al.  Improved anticontaminator for cryo‐electron microscopy with a Philips EM 400 , 1984 .

[16]  J. Dubochet,et al.  Cryo-electron microscopy of vitrified specimens , 1988, Quarterly Reviews of Biophysics.

[17]  R. Henderson,et al.  Model for the structure of bacteriorhodopsin based on high-resolution electron cryo-microscopy. , 1990, Journal of molecular biology.

[18]  R. Schröder,et al.  Zero-loss energy filtering as improved imaging mode in cryoelectronmicroscopy of frozen-hydrated specimens☆ , 1990 .

[19]  W. Chiu,et al.  High resolution cryo system designed for JEM 100CX electron microscope. , 1987, Ultramicroscopy.

[20]  K A Taylor,et al.  Electron microscopy of frozen hydrated biological specimens. , 1976, Journal of ultrastructure research.

[21]  R. Milligan,et al.  Molecular structure determination of crystalline specimens in frozen aqueous solutions. , 1984, Ultramicroscopy.

[22]  R A Crowther,et al.  Visualization of alpha-helices in tobacco mosaic virus by cryo-electron microscopy. , 1989, Journal of molecular biology.

[23]  R Henderson,et al.  Image contrast in high-resolution electron microscopy of biological macromolecules: TMV in ice. , 1992, Ultramicroscopy.

[24]  D. Stokes,et al.  Structure of CaATPase: electron microscopy of frozen-hydrated crystals at 6 A resolution in projection. , 1990, Journal of molecular biology.

[25]  N. Unwin.  Nicotinic acetylcholine receptor at 9 A resolution. , 1993, Journal of molecular biology.

[26]  F. Franks.  The Properties of Aqueous Solutions at Subzero Temperatures , 1982 .

[27]  W. Kühlbrandt,et al.  High-resolution electron crystallography of light-harvesting chlorophyll a/b-protein complex in three different media. , 1991, Journal of molecular biology.

[28]  K. Adachi,et al.  A new method of preparation of a self-perforated micro plastic grid and its application. , 1965, Journal of electron microscopy.

[29]  R. Henderson,et al.  Images of purple membrane at 2.8 A resolution obtained by cryo-electron microscopy. , 1988, Journal of molecular biology.