Quantitative analysis of low-dose high-resolution images and diffraction patterns from zeolites using a slow-scan CCD camera

1993 ◽  
Vol 51 ◽  
pp. 670-671
Author(s):  
M. Pan ◽  
P.A. Crozier
Keyword(s):  
High Resolution ◽  
Molecular Sieves ◽  
Structural Damage ◽  
Low Dose ◽  
Signal To Noise Ratio ◽  
Ccd Camera ◽  
Real Space ◽  
High Signal ◽  
Resolution Electron ◽  
Diffraction Patterns

Zeolites are an important class of low-density aluminosilicates framework structures with applications to the field of catalysis and molecular sieves. In order to understand die origin of die unique properties of these materials and hence optimize their performance, it is essential to have a detailed description of their structures. Zeolite structure is often described in terms of the secondary building unit (SBU) which is a specific configuration of the SiO4 tetrahedrons. Structure determination of zeolites is then reduced to determine the types of SB Us and their connectivities when forming the 3-D framework structure.It has been recognized that zeolites undergo rapid structural damage under electron beam irradiation. The use of a slow-scan CCD camera to record low-dose high resolution electron microscope (HREM) images from zeolites with the combination with real-space averaging techniques has been shown to be successful in obtaining the averaged unit cell with high signal-to-noise ratio (SNR).

Slow-scan CCD cameras and low-dose High-Resolution Electron Microscopy: Direct and quantitative

1994 ◽  
Vol 52 ◽  
pp. 412-413
Author(s):  
M. Pan
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Structural Damage ◽  
Low Dose ◽  
Dynamic Range ◽  
High Resolution Electron Microscopy ◽  
Operating Conditions ◽  
Digital Data ◽  
Ccd Cameras ◽  
Resolution Electron

It has been known for many years that materials such as zeolites, polymers, and biological specimens have crystalline structures that are vulnerable to electron beam irradiation. This radiation damage severely restrains the use of high resolution electron microscopy (HREM). As a result, structural characterization of these materials using HREM techniques becomes difficult and challenging. The emergence of slow-scan CCD cameras in recent years has made it possible to record high resolution (∽2Å) structural images with low beam intensity before any apparent structural damage occurs. Among the many ideal properties of slow-scan CCD cameras, the low readout noise and digital recording allow for low-dose HREM to be carried out in an efficient and quantitative way. For example, the image quality (or resolution) can be readily evaluated on-line at the microscope and this information can then be used to optimize the operating conditions, thus ensuring that high quality images are recorded. Since slow-scan CCD cameras output (undistorted) digital data within the large dynamic range (103-104), they are ideal for quantitative electron diffraction and microscopy.

Characteristic feature on structure analyzed by high-resolution electron crystallography

1995 ◽  
Vol 53 ◽  
pp. 834-835
Author(s):  
Y. Fujiyoshi ◽  
K. Mitsuoka ◽  
T. Hirai ◽  
K. Murata ◽  
A. Miyazawa ◽  
...  
Keyword(s):  
Amino Acid ◽  
High Resolution ◽  
Characteristic Feature ◽  
Proton Pump ◽  
Ccd Camera ◽  
Electron Crystallography ◽  
Embedding Technique ◽  
Resolution Electron ◽  
Ph Conditions ◽  
Diffraction Patterns

The structure of bacteriorhodopsin (bR), which was already analyzed by Henderson et al., is studied by our new electron cryo-microscope equipped with Field Emission Gun (FEG) and Slow Scan CCD camera (SSCCD), because our system together with ice embedding technique enable us to solve the structure of bR at various pH conditions between pH 4.0 and 10.0. Ionization of amino acid is naturally closely related to the translocation of proton and then the function of the proton pump of bR. Therefore, observation of translocation of proton in bR is very important, if possible. Both ice embedding and high resolution techniques are essential to achieve this intention. Therefore, we intended to develop an electron cryo-microscope fit to these techniques and recently we had succeeded it.We collected whole sets of diffraction patterns for bR up to 70 degree tilt at pH 5.5 by using SSCCD, and merged these data of 300 diffraction patterns.

scholarly journals Seismology of southern Be stars

1994 ◽  
Vol 162 ◽  
pp. 104-105
Author(s):  
Eduardo Janot-Pacheco ◽  
Nelson Vani Leister
Keyword(s):  
High Resolution ◽  
Signal To Noise Ratio ◽  
Ccd Camera ◽  
High Signal ◽  
Be Stars ◽  
Signal To Noise ◽  
Photometric Variability ◽  
Spectroscopic Observations ◽  
Photometric Observations ◽  
Noise Ratio

We have started in 1990 a search for moving bumps in the HeI λ 667.8 nm of mainly southern, bright Be stars. The objects of our sample have been selected on the basis of photometric variability (Cuypers et al., 1989). High resolution (R≥ 30,000), high signal-to-noise ratio (S/R≥ 300) spectroscopic observations have been performed at the brazilian Laboratório Nacional de Astrofísica with a CCD camera attached to the coudé spectrograph of the 1.60 m telescope (e.g. Table I). Several hundred spectra have been taken during the last three years. Photometric observations simultaneous with spectroscopy were made on the same site in July 1992 with a two-channel photometer (Stromgren b filter) and a CCD camera (Johnson B filter) installed at two 0.60 m telescopes. The idea is try to disentangle the controversy between NRP and RM models with the help of simultaneous spectroscopy and photometry.

Cooling-induced wrinkling of thin crystals of biological macromolecules can be prevented by using molybdenum grids

1992 ◽  
Vol 50 (1) ◽  
pp. 520-521
Author(s):  
Robert M. Glaeser
Keyword(s):  
High Resolution ◽  
Signal To Noise Ratio ◽  
Data Retrieval ◽  
Biological Macromolecules ◽  
Tilt Axis ◽  
Resolution Electron ◽  
High Resolution Images ◽  
Diffraction Patterns ◽  
Rate Of Success ◽  
High Degree

It is an important requirement of high resolution electron diffraction and electron microscopy of thin crystals of biological macromolecules that the specimen be flat (i.e. planar) to one degree or less over distances of one micrometer or more. This high degree of specimen flatness is required in order to collect diffraction patterns and images at high tilt angles and high resolution. Imperfect flatness causes the diffraction spots which are perpendicular to the tilt axis to become streaked or blurred, while the spots parallel to the tilt axis remain sharp. If the specimen wrinkling, or mosaic angular spread, is too severe, the diffraction spots overlap one another to make a continuum, and data retrieval becomes impossible (see Figure 3, for an example). Even before that point is reached, the broadening of the diffraction spots makes it increasingly difficult to obtain accurate background-subtracted diffraction intensities, and the signal-to-noise ratio in the computed Fourier transform of high resolution images is severely decreased. In some preparations of thin protein crystals the stringent requirements for specimen flatness can be met with a reasonable rate of success. In the case of bacteriorhodopsin, prepared as large singlecrystal sheets by detergent-induced fusion of the native purple membrane of Halobacterium halobium, the frequency of success is normally very low.

Low dose electron diffraction of wet protein crystals

1978 ◽  
Vol 36 (2) ◽  
pp. 6-7
Author(s):  
B.B. Chang ◽  
D.F. Parsons
Keyword(s):  
High Resolution ◽  
Electron Diffraction ◽  
Radiation Damage ◽  
Low Dose ◽  
Protein Crystal ◽  
Protein Crystals ◽  
Electron Dose ◽  
Resolution Electron ◽  
Diffraction Patterns ◽  
Iterative Procedures

High resolution electron diffraction patterns from wet unstained protein-crystals have been successfully obtained by using very low electron dose (∽10-2 e/Å2, much smaller dose than used by Unwin and Henderson (1975) for electron diffraction of their crystals). This enabled us to follow the radiation damage of the protein crystal due to increasing dosage by recording successive diffraction patterns given by the same crystal. Changes in intensities of both the high and the low order reflections can be studied. Another important consequence is that very low dose electron diffraction can be obtained from the same crystal and iterative procedures such as Gerchberg and Saxton's technique can be applied.The electron diffraction work with very low dose was performed at 200 kV using the environmental chamber in the Jeolco 200. To achieve best electron diffraction conditions with the specimen in the hydration chamber, the objective and intermediate lens currents have been changed.

Light Optical Diffraction Studies of Macromolecular Ultrastructure

1970 ◽  
Vol 28 ◽  
pp. 258-259
Author(s):  
Glen B. Haydon
Keyword(s):  
High Resolution ◽  
Diffraction Analysis ◽  
Diffraction Pattern ◽  
Electron Micrographs ◽  
Optical Diffraction ◽  
Electron Micrograph ◽  
Its Analysis ◽  
Resolution Electron ◽  
Diffraction Patterns

Analysis of light optical diffraction patterns produced by electron micrographs can easily lead to much nonsense. Such diffraction patterns are referred to as optical transforms and are compared with transforms produced by a variety of mathematical manipulations. In the use of light optical diffraction patterns to study periodicities in macromolecular ultrastructures, a number of potential pitfalls have been rediscovered. The limitations apply to the formation of the electron micrograph as well as its analysis.(1) The high resolution electron micrograph is itself a complex diffraction pattern resulting from the specimen, its stain, and its supporting substrate. Cowley and Moodie (Proc. Phys. Soc. B, LXX 497, 1957) demonstrated changing image patterns with changes in focus. Similar defocus images have been subjected to further light optical diffraction analysis.

Structural analysis of the surface-layer protein of spirillum serpens by high-resolution electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 738-739
Author(s):  
W. H. Wu ◽  
R. M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
Surface Layer ◽  
Structural Analysis ◽  
High Resolution ◽  
Low Dose ◽  
High Resolution Electron Microscopy ◽  
Optical Diffraction ◽  
Surface Layer Protein ◽  
Morphological Unit ◽  
Resolution Electron

Spirillum serpens possesses a surface layer protein which exhibits a regular hexagonal packing of the morphological subunits. A morphological model of the structure of the protein has been proposed at a resolution of about 25 Å, in which the morphological unit might be described as having the appearance of a flared-out, hollow cylinder with six ÅspokesÅ at the flared end. In order to understand the detailed association of the macromolecules, it is necessary to do a high resolution structural analysis. Large, single layered arrays of the surface layer protein have been obtained for this purpose by means of extensive heating in high CaCl2, a procedure derived from that of Buckmire and Murray. Low dose, low temperature electron microscopy has been applied to the large arrays.As a first step, the samples were negatively stained with neutralized phosphotungstic acid, and the specimens were imaged at 40,000 magnification by use of a high resolution cold stage on a JE0L 100B. Low dose images were recorded with exposures of 7-9 electrons/Å2. The micrographs obtained (Fig. 1) were examined by use of optical diffraction (Fig. 2) to tell what areas were especially well ordered.

Correlation averaging of two-dimensional crystals: basic strategy and refinements

1986 ◽  
Vol 44 ◽  
pp. 136-139
Author(s):  
W. Baumeister ◽  
R. Rachel ◽  
R. Guckenberger ◽  
R. Hegerl
Keyword(s):  
Low Dose ◽  
Signal To Noise Ratio ◽  
Real Space ◽  
Fourier Domain ◽  
Two Dimensional ◽  
Signal To Noise ◽  
Unit Cells ◽  
Lateral Displacements ◽  
Established Technique ◽  
Crystalline Array

IntroductionCorrelation averaging (CAV) is meanwhile an established technique in image processing of two-dimensional crystals /1,2/. The basic idea is to detect the real positions of unit cells in a crystalline array by means of correlation functions and to average them by real space superposition of the aligned motifs. The signal-to-noise ratio improves in proportion to the number of motifs included in the average. Unlike filtering in the Fourier domain, CAV corrects for lateral displacements of the unit cells; thus it avoids the loss of resolution entailed by these distortions in the conventional approach. Here we report on some variants of the method, aimed at retrieving a maximum of information from images with very low signal-to-noise ratios (low dose microscopy of unstained or lightly stained specimens) while keeping the procedure economical.

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