Effects of illumination spot size on high-resolution image contrast for radiation-sensitive specimens

Author(s):  
Kenneth H. Downing

Electron diffraction patterns of a number of different protein crystals extend to well beyond 0.4 nm. However, until quite recently, no images of these crystals had been obtained which showed such high resolution. The recent introduction of monolayer crystals of paraffin, which diffract at 0.4 nm several thousand times as strongly as typical protein crystals, has made it possible to obtain such high-resolution images almost routinely, and has allowed the study of the causes for the previous lack of success. It has been found that the images of paraffin crystals fall far short of images which could be obtained under ideal circumstances. Not only do the images only rarely show the pseudo-hexagonal symmetry of the crystals, but quantitative analysis of lattice images has shown that under normal conditions the image contrast is typically only about 3-4% of that which is theoretically possible, based on the strength of electron diffraction spots.

2001 ◽  
Vol 16 (1) ◽  
pp. 101-107 ◽  
Author(s):  
Takeo Oku ◽  
Jan-Olov Bovin ◽  
Iwami Higashi ◽  
Takaho Tanaka ◽  
Yoshio Ishizawa

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.


Author(s):  
Kenneth H. Downing

Electron microscopists have for some time been progressing toward the goal of atomic resolution of the structure of several proteins; these are proteins that are available in thin crystals that can produce electron diffraction patterns extending beyond 3 Å. The diffraction patterns demonstrate that the specimens themselves should be amenable to high resolution studies. Progress in the solution of these structures has been limited, however, because it has not yet been possible to obtain images of the crystals which have the same quality as the diffraction patterns. Fourier transforms of the images show diffraction spots which decrease in intensity with increasing resolution much faster than do the corresponding electron diffraction patterns, recorded under essentially the same conditions.The source of this limitation in image quality is clearly not the electron microscope. Materials scientists routinely achieve the resolution which structural biologists still strive for.


Author(s):  
J. Brink ◽  
W. Chiu

Images of radiation-sensitive two-dimensional crystals quite often fail to display the same resolution visible in electron diffraction patterns of these crystals. It is now generally accepted that besides the microscope's envelope functions, radiation damage and the MTF of the film and cryo-holder, beam-induced specimen motion is a major contrast degrading factor. Minimization of this movement proved possible through use of narrow electron beams, i.e. in spot-scanning. Routine high resolution imaging, however, remains a difficult task. We have investigated the possibilities of enhancing the effeciency this goal by using 400 keV electrons. Based upon overall less attenuation from the envelope functions at 400 kV, structure factors at around 3Å resolution would show amplitudes easily twice as large as compared to 100 kV. Further, at 400 kV inelastic scattering would be reduced relative to 100 kV. Moreover, it has been suggested that image contrast would increase roughly proportional to β. Additional advantages concerning for instance Ewald sphere curvature and depth of field have been put forward in Chiu et al.


1991 ◽  
Vol 231 ◽  
Author(s):  
L. Salamanca Riba ◽  
K. Park ◽  
B. T. Jonker

AbstractWe have observed an ordered structure in Zn0.5Fe0.5Se epilayers grown on (001) InP substrates using transmission electron microscopy. The ordered structure of Zn0.5Fe0.5Se has Fe atoms occupying the (0,0,0) and (½, ½, 0) sites and Zn atoms occupying the (0, ½, ½) and (½, 0, ½) sites in the zinc-blende unit cell. Ordering is observed in both electron diffraction patterns and cross-sectional high-resolution lattice images along the < 100 > and < 110 > directions. This ordered structure consists of alternating ZnSe and FeSe monolayers along the < 100 > and < 110 > directions. Computer image simulations of the high-resolution images under various thicknesses, and defocusing conditions have been obtained and are compared with those obtained experimentally.


2012 ◽  
Vol 186 ◽  
pp. 255-258 ◽  
Author(s):  
Katarzyna Stan ◽  
Lidia Lityńska-Dobrzyńska ◽  
Jan Dutkiewicz ◽  
Lukasz Rogal ◽  
Anna Maria Janus

Microstructure of rapidly solidified Al91Mn7Fe2 (at.%) alloy was investigated using SEM and TEM techniques. Quasicrystalline particles of different shapes and sizes embedded in the aluminium matrix were observed. Quasilattice constant was calculated as 0.461 Å. Additionally orientation relationships between matrix and quasicrystals particles were found based on electron diffraction patterns and high resolution images, such that: five-fold axis lie along [011] or [001] axes of the α-Al crystallographic direction.


1995 ◽  
Vol 10 (6) ◽  
pp. 1539-1545 ◽  
Author(s):  
G.Z. Pan ◽  
A. Michel ◽  
V. Pierron-Bohnes ◽  
P. Vennéguès ◽  
M.C. Cadeville

Plan-view microstructures of two Co/Ru bilayers with a composition of [Co12ÅRu45Å]2 and [Co40ÅRu35Å]2 have been studied by conventional and high resolution electron microscopy. Large differences in electron diffraction and image contrast between the two bilayers were observed, which are recognized as the microstructural variations during the relaxation of large coherent planar strains when the Co layers wet coherently or semicoherently the Ru layers. For the [Co12ÅRu45Å]2 bilayer, the Co layers are unrelaxed from the Ru layers; only one set of electron diffraction patterns was observed, and the image consists of three types of contrasts which are closely related with either the generation and movement of misfit dislocations or large coherent strains. For the [Co40ÅRu35Å]2 bilayer, the Co layers are relaxed basically from the Ru layers; two sets of electron diffraction patterns with double diffraction spots were observed, and the image consists of small irregular areas with moiré fringe dots.


2007 ◽  
Vol 336-338 ◽  
pp. 818-821
Author(s):  
Kunio Yubuta ◽  
Satoshi Okada ◽  
Yuzuru Miyazaki ◽  
Ichiro Terasaki ◽  
Tsuyoshi Kajitani

We have investigated the modulated structure of the misfit-layered crystal Bi1.8Sr2.0Rh1.6Ox by means of electron diffraction and high-resolution electron microscopy. This compound consists of two interpenetrating subsystems of a hexagonal RhO2 sheet and a distorted four-layered rock-salt-type (Bi,Sr)O block. Both subsystems have common a-, c-axes and β-angles with a = 5.28 Å, c = 29.77 Å and β = 93.7º. On the other hand, the crystal structure is incommensurated parallel to the b-axes, among which b1 = 3.07 Å for the RhO2 sheet and b2 = 4.88 Å for the (Bi,Sr)O block. The misfit ratio, b1/b2 ~ 0.63, characterizes the structural analogue as [Bi1.79Sr1.98Oy]0.63[RhO2]. This compound has two modulation vectors, i.e., q1 = – a* + 0.63b1* and q2 = 0.17b1* + c*, and the superspace group is assigned as the Cc(1β0, 0μ1)-type from the electron diffraction patterns. High-resolution images taken with the incident electron beam parallel to the a- and c-axes clearly show displacive as well as compositional modulations.


Author(s):  
B.B. Chang ◽  
D.F. Parsons

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.


2000 ◽  
Vol 56 (3) ◽  
pp. 363-368 ◽  
Author(s):  
Takeo Oku ◽  
Anna Carlsson ◽  
Jan-Olov Bovin ◽  
Christer Svensson ◽  
L. Reine Wallenberg ◽  
...  

The modulated structure of Ag2SnO3, disilver tin trioxide, was investigated by high-resolution electron microscopy and electron diffraction along four different directions. Electron diffraction showed an incommensurate one-dimensional modulated structure with a modulation wavevector of 1/6.4a*. High-resolution images showed a large number of superstructure domains with the size range 10–100 nm and orientations related by hexagonal rotation. The modulation was determined to be displacements along the c axis of the Ag atoms both in octahedral and linear coordination. An approximate structure model with a commensurate sixfold superstructure, with an orthorhombic cell (P212121, a = 2.922, b = 1.267, c = 0.562 nm), is proposed. Calculated images and electron diffraction patterns, based on this model, agree well with experimental observations.


Author(s):  
Etienne de Harven

Biological ultrastructures have been extensively studied with the scanning electron microscope (SEM) for the past 12 years mainly because this instrument offers accurate and reproducible high resolution images of cell shapes, provided the cells are dried in ways which will spare them the damage which would be caused by air drying. This can be achieved by several techniques among which the critical point drying technique of T. Anderson has been, by far, the most reproducibly successful. Many biologists, however, have been interpreting SEM micrographs in terms of an exclusive secondary electron imaging (SEI) process in which the resolution is primarily limited by the spot size of the primary incident beam. in fact, this is not the case since it appears that high resolution, even on uncoated samples, is probably compromised by the emission of secondary electrons of much more complex origin.When an incident primary electron beam interacts with the surface of most biological samples, a large percentage of the electrons penetrate below the surface of the exposed cells.


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