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.

Multiple isomorphous replacement for phase determination in protein crystals

1995 ◽  
Vol 53 ◽  
pp. 856-857
Author(s):  
Christoph Burmester ◽  
Kenneth C. Holmes ◽  
Rasmus R. Schröder
Keyword(s):  
Electron Diffraction ◽  
Heavy Atom ◽  
Atomic Resolution ◽  
Protein Crystals ◽  
Electron Diffraction Data ◽  
Phase Information ◽  
Electron Dose ◽  
Isomorphous Replacement ◽  
Resolution Electron ◽  
Diffraction Patterns

Electron crystallography of 2D protein crystals can yield models with atomic resolution by taking Fourier amplitudes from electron diffraction and phase information from processed images. Imaging at atomic resolution is more difficult than the recording of corresponding high resolution electron diffraction patterns. Therefore attempts have been made to retrieve phase information from diffraction from heavy atom labelled protein crystals. The expected differences between native and labelled crystals are small, therefore a high experimental accuracy is necessary. This is achieved by the use of energy filter TEM and image plates, as dicussed in. Here we present electron diffraction data obtained from frozen hydrated 3D protein crystals with an energy filter microspcope and a specially designed image plate scanner. Data were recorded for the native crystal as well as for two different heavy atom derivatives. Differences between the native and the derivate forms can be detected and are significant.Electron diffraction patterns from frozen hydrated catalase crystals were recorded on an EFTEM Zeiss 912 Ω (120kV, zero loss mode, energy width ΔE=10eV, electron dose 5 e-/A2) using image plates and a quasi confocal scanner readout.

Anisotropic radiation damage of potassium tetracyano platinate (KCP)

1978 ◽  
Vol 36 (1) ◽  
pp. 392-393
Author(s):  
N. Uyeda ◽  
E. J. Kirkland ◽  
B. M. Siegel
Keyword(s):  
Electron Diffraction ◽  
Radiation Damage ◽  
Structural Changes ◽  
High Resolution Electron Microscopy ◽  
Radiation Sensitivity ◽  
Resolution Electron ◽  
Damage Process ◽  
Diffraction Patterns ◽  
Fading Rate

The direct observation of structural change by high resolution electron microscopy will be essential for the better understanding of the damage process and its mechanism. However, this approach still involves some difficulty in quantitative interpretation mostly being due to the quality of obtained images. Electron diffraction, using crystalline specimens, has been the method most frequently applied to obtain a comparison of radiation sensitivity of various materials on the quantitative base. If a series of single crystal patterns are obtained the fading rate of reflections during the damage process give good comparative measures. The electron diffraction patterns also render useful information concerning the structural changes in the crystal. In the present work, the radiation damage of potassium tetracyano-platinate was dealt with on the basis two dimensional observation of fading rates of diffraction spots. KCP is known as an ionic crystal which possesses “one dimensional” electronic properties and it would be of great interest to know if radiation damage proceeds in a strongly asymmetric manner.

Atomic structure of YB56 studied by digital high-resolution electron microscopy and electron diffraction

2001 ◽  
Vol 16 (1) ◽  
pp. 101-107 ◽  
Author(s):  
Takeo Oku ◽  
Jan-Olov Bovin ◽  
Iwami Higashi ◽  
Takaho Tanaka ◽  
Yoshio Ishizawa
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Electron Diffraction ◽  
High Resolution Electron Microscopy ◽  
Charge Coupled Device ◽  
X Ray ◽  
Resolution Electron ◽  
High Resolution Images ◽  
Diffraction Patterns ◽  
Simulated Images

Atomic positions for Y atoms were determined by using high-resolution electron microscopy and electron diffraction. A slow-scan charge-coupled device camera which had high linearity and electron sensitivity was used to record high-resolution images and electron diffraction patterns digitally. Crystallographic image processing was applied for image analysis, which provided more accurate, averaged Y atom positions. In addition, atomic disordering positions in YB56 were detected from the differential images between observed and simulated images based on x-ray data, which were B24 clusters around the Y-holes. The present work indicates that the structure analysis combined with digital high-resolution electron microscopy, electron diffraction, and differential images is useful for the evaluation of atomic positions and disordering in the boron-based crystals.

Small-Spot Illumination For High-Resolution Electron Microscopy of Beam-Sensitive Specimens

1985 ◽  
Vol 43 ◽  
pp. 320-321
Author(s):  
Kenneth H. Downing ◽  
Robert M. Glaeser
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
High Resolution ◽  
Electron Diffraction ◽  
High Resolution Electron Microscopy ◽  
Photographic Emulsion ◽  
Lateral Motion ◽  
Small Spot ◽  
Resolution Electron ◽  
Diffraction Patterns

The contrast observed in images of beam-sensitive, crystalline specimens is found to be significantly less than one would predict based on observations of electron diffraction patterns of the specimens. Factors such as finite coherence, inelastic scattering, and the limited MTF of the photographic emulsion account for some decrease in contrast. It appears, however, that most of the loss in signal is caused by motion of the specimen during exposure to the electron beam. The introduction of point and other defects in the crystal, resulting from radiation damage, causes bending and lateral motion, which degrade the contrast in the image. We have therefore sought to determine whether the beam-induced specimen motion can be reduced by reducing the area of the specimen which is illuminated at any one time.

On the interpretation of electron diffraction patterns from materials containing planar boundaries: the intergrowth tungsten bronzes

1990 ◽  
Vol 429 (1876) ◽  
pp. 91-117 ◽  
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Electron Diffraction ◽  
High Resolution Electron Microscopy ◽  
Optical Diffraction ◽  
General Interest ◽  
Transmission Electron ◽  
Resolution Electron ◽  
Resolution Imaging ◽  
Diffraction Patterns

Materials containing planar boundaries are of general interest and complete understanding of their structures is important. When direct imaging of the boundaries by, for instance, high-resolution electron microscopy, is impracticable, details of their structure and arrangement may be obtained from electron diffraction patterns. Such patterns are discussed in terms of those from intergrowth tungsten bronzes as specific examples. Fourier-transform calculations for proposed structures have been made to establish, in conjunction with optical-diffraction analogues, the features of the far-field diffraction patterns. These results have been compared with diffraction patterns obtained experimentally by transmission electron microscopy. The aim of the study, to show that the arrangement of the boundaries in these complicated phases can be deduced from their diffraction patterns without the need for high-resolution imaging, has been achieved. The steps to be taken to make these deductions are set out.

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).

Application of the imaging plate to quantitative analysis of electron-diffraction and high-resolution electron microscopy

1989 ◽  
Vol 47 ◽  
pp. 666-667
Author(s):  
K. Hiraga ◽  
D. Shindo ◽  
M. Hirabayashi ◽  
T. Oikawa ◽  
N. Mori ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Quantitative Analysis ◽  
High Resolution ◽  
Electron Diffraction ◽  
Electron Micrographs ◽  
High Resolution Electron Microscopy ◽  
Imaging Plate ◽  
Resolution Electron ◽  
Diffraction Patterns ◽  
High Resolution Electron Micrographs

The “Imaging Plate” (IP) has three superior characteristics, i.e., high sensitivity to the electron beam, and a wide dynamic range and good linearity for electron dose compared with conventional EM films. The use of the IP is expected to lead to quantitative analysis of electron microscopy. The purpose of the present work is to examine the possibility of application of the IP to the quantitative analysis of electron diffraction and high-resolution electron microscopy.By using the TEM-IP System developed by Oikawa et al., which is published in this conference, electron diffraction patterns and high-resolution electron micrographs taken on the IP with an effective size of 102 х 77 mm2 were convertedinto digital data of 2048 х 1536 pixels with 4096 gray levels. Observations of electron diffraction patterns and high-resolution electron micrographs were made with a 200 kV (JEM-2000FX) and a 400 kV (JEM-4000EX) electron microscope, respectively.

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