Improvements in electron diffraction of frozen hydrated crystals by energy filtering and large-area single-electron detection

Author(s):  
Rasmus R. Schröder ◽  
Christoph Burmester

Diffraction patterns of 3D protein crystals embedded in vitrious ice are critical to record. Inelastically scattered electrons almost completely superimpose the diffraction pattern of crystals if the thickness of the crystal is higher than the mean free path of electrons in the specimen. Figure 1 shows such an example of an unfiltered electron diffraction pattern from a frozen hydrated 3D catalase crystal. However, for thin 2D crystals electron diffraction has been the state of the art method to determine the Fourier amplitudes for reconstructions to atomic level, and in one case the possibility of obtaining Fourier phases from diffraction patterns has been studied. One of the main problems could be the background in the diffraction pattern due to inelastic scattering and the recording characteristics for electrons of conventional negative material.It was pointed out before, that the use of an energy filtered TEM (EFTEM) and of the Image Plate as a large area electron detector gives considerable improvement for detection of diffraction patterns.

Author(s):  
G. Lehmpfuhl

Introduction In electron microscopic investigations of crystalline specimens the direct observation of the electron diffraction pattern gives additional information about the specimen. The quality of this information depends on the quality of the crystals or the crystal area contributing to the diffraction pattern. By selected area diffraction in a conventional electron microscope, specimen areas as small as 1 µ in diameter can be investigated. It is well known that crystal areas of that size which must be thin enough (in the order of 1000 Å) for electron microscopic investigations are normally somewhat distorted by bending, or they are not homogeneous. Furthermore, the crystal surface is not well defined over such a large area. These are facts which cause reduction of information in the diffraction pattern. The intensity of a diffraction spot, for example, depends on the crystal thickness. If the thickness is not uniform over the investigated area, one observes an averaged intensity, so that the intensity distribution in the diffraction pattern cannot be used for an analysis unless additional information is available.


Author(s):  
Werner Kühlbrandt ◽  
Da Neng Wang ◽  
K.H. Downing

The light-harvesting chlorophyll-a/b protein complex (LHC-II) is the most abundant membrane protein in the chloroplasts of green plants where it functions as a molecular antenna of solar energy for photosynthesis. We have grown two-dimensional (2d) crystals of the purified, detergent-solubilized LHC-II . The crystals which measured 5 to 10 μm in diameter were stabilized for electron microscopy by washing with a 0.5% solution of tannin. Electron diffraction patterns of untilted 2d crystals cooled to 130 K showed sharp spots to 3.1 Å resolution. Spot-scan images of 2d crystals were recorded at 160 K with the Berkeley microscope . Images of untilted crystals were processed, using the unbending procedure by Henderson et al . A projection map of the complex at 3.7Å resolution was generated from electron diffraction amplitudes and high-resolution phases obtained by image processing .A difference Fourier analysis with the same image phases and electron diffraction amplitudes recorded of frozen, hydrated specimens showed no significant differences in the 3.7Å projection map. Our tannin treatment therefore does not affect the structural integrity of the complex.


2000 ◽  
Vol 33 (5) ◽  
pp. 1246-1252 ◽  
Author(s):  
Elizabeth J. Grier ◽  
Amanda K. Petford-Long ◽  
Roger C. C. Ward

Computer simulations of the electron diffraction patterns along the [\bar{1}10] zone axes of four ordered structures within the β-RH2+xphase, withR= Ho or Y, and 0 ≤x≤ 0.25, have been performed to establish whether or not the hydrogen ordering could be detected using electron diffraction techniques. Ordered structures within otherRH2+x(R= Ce, Tb) systems have been characterized with neutron scattering experiments; however, for HoH(D)2+x, neutron scattering failed to characterize the superstructure, possibly because of the lowxconcentration or lack of long-range order within the crystal. This paper aims to show that electron diffraction could overcome both of these problems. The structures considered were the stoichiometric face-centred cubic (f.c.c.) fluorite structure (x= 0), theD1 structure (x= 0.125), theD1astructure (x= 0.2) and theD022structure (x= 0.25). In the stoichiometric structure, with all hydrogen atoms located on the tetrahedral (t) sites, only the diffraction pattern from the f.c.c. metal lattice was seen; however, for the superstoichiometric structures, with the excess hydrogen atoms ordered on the octahedral (o) sites, extra reflections were visible. All the superstoichiometric structures showed extra reflections at the (001)f.c.c.and (110)f.c.c.type positions, with structureD1 also showing extra peaks at (½ ½ ½)f.c.c.. These reflections are not seen in the simulations at similar hydrogen concentrations with the hydrogen atoms randomly occupying theovacancies.


2007 ◽  
Vol 13 (5) ◽  
pp. 329-335 ◽  
Author(s):  
Suk Chung ◽  
David J. Smith ◽  
Martha R. McCartney

The mean-free-paths for inelastic scattering of high-energy electrons (200 keV) for AlAs and GaAs have been determined based on a comparison of thicknesses as measured by electron holography and convergent-beam electron diffraction. The measured values are 77 ± 4 nm and 67 ± 4 nm for AlAs and GaAs, respectively. Using these values, the mean inner potentials of AlAs and GaAs were then determined, from a total of 15 separate experimental measurements, to be 12.1 ± 0.7 V and 14.0 ± 0.6 V, respectively. These latter measurements show good agreement with recent theoretical calculations within experimental error.


1993 ◽  
Vol 52 (3-4) ◽  
pp. 454-458 ◽  
Author(s):  
R. Holmestad ◽  
O.L. Krivanek ◽  
R. Høier ◽  
K. Marthinsen ◽  
J.C.H. Spence

1997 ◽  
Vol 3 (S2) ◽  
pp. 973-974
Author(s):  
A.G. Fox ◽  
E.S.K. Menon ◽  
M. Saunders

Over the last ten years TEMs have been developed that are capable of HREM, EDX, PEELS and diffraction using a single objective pole piece. More recently these TEMs have been equipped with the capability of energy filtering the electron beam after it has passed through the sample so that energy filtered images and electron diffraction patterns can be obtained. In this work the use of a Topcon 002B TEM equipped with a GATAN PEELS imaging filter (GIF) to generate zero-loss energy filtered zone axis CBED patterns and elemental images from inelastically scattered electrons will be described. An analysis of this energy filtered data indicates that elemental imaging using the GIF is an informative, but semiquantitative technique, whereas zero-loss energy filtered zone axis CBED patterns can be accurately quantified so that the two lowest-angle x-ray form factors of cubic elements can be measured with errors of the order of 0.1% or less.


1988 ◽  
Vol 32 ◽  
pp. 593-600
Author(s):  
J. C. Russ ◽  
T. Taguchi ◽  
P. M. Peters ◽  
E. Chatfield ◽  
J. C. Russ ◽  
...  

Conventional selected area diffraction patterns as obtained in the TEM present difficulties for identification of materials such as asbestifonn minerals, although diffraction data is considered to be one of the preferred methods for making this identification. The preferred orientation of the fibers in each field of measurement, and the spotty patterns that are obtained, do not readily lend themselves to measurement of the integrated intensity values for each dspacing, and even the d-spacings may be hard to determine precisely because the true center location for the broken rings requires estimation. To overcome these problems, we have implemented an automatic method for diffraction pattern measurement. It automatically locates the center of patterns with high precision, measures the radius of each ring of spots in the pattern, and integrates the density of spots in that ring.


1994 ◽  
Vol 9 (9) ◽  
pp. 2450-2456 ◽  
Author(s):  
L.C. Qin

Electron diffraction intensities from cylindrical objects can be conveniently analyzed using Bessel functions. Analytic formulas and geometry of the diffraction patterns from cylindrical carbon nanotubes are presented in general forms in terms of structural parameters, such as the pitch angle and the radius of a tubule. As an example the Fraunhofer diffraction pattern from a graphitic tubule of structure [18,2] has been simulated to illustrate the characteristics of such diffraction patterns. The validity of the projection approximation is also discussed.


Nanomaterials ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 962
Author(s):  
Miroslav Slouf ◽  
Radim Skoupy ◽  
Ewa Pavlova ◽  
Vladislav Krzyzanek

We introduce a novel scanning electron microscopy (SEM) method which yields powder electron diffraction patterns. The only requirement is that the SEM microscope must be equipped with a pixelated detector of transmitted electrons. The pixelated detectors for SEM have been commercialized recently. They can be used routinely to collect a high number of electron diffraction patterns from individual nanocrystals and/or locations (this is called four-dimensional scanning transmission electron microscopy (4D-STEM), as we obtain two-dimensional (2D) information for each pixel of the 2D scanning array). Nevertheless, the individual 4D-STEM diffractograms are difficult to analyze due to the random orientation of nanocrystalline material. In our method, all individual diffractograms (showing randomly oriented diffraction spots from a few nanocrystals) are combined into one composite diffraction pattern (showing diffraction rings typical of polycrystalline/powder materials). The final powder diffraction pattern can be analyzed by means of standard programs for TEM/SAED (Selected-Area Electron Diffraction). We called our new method 4D-STEM/PNBD (Powder NanoBeam Diffraction) and applied it to three different systems: Au nano-islands (well diffracting nanocrystals with size ~20 nm), small TbF3 nanocrystals (size < 5 nm), and large NaYF4 nanocrystals (size > 100 nm). In all three cases, the STEM/PNBD results were comparable to those obtained from TEM/SAED. Therefore, the 4D-STEM/PNBD method enables fast and simple analysis of nanocrystalline materials, which opens quite new possibilities in the field of SEM.


Author(s):  
A. G. Jackson ◽  
M. Rowe

The analysis of electron diffraction patterns can be done automatically using image techniques for collecting the diffraction pattern. This is useful if the material is well known. The alternative when the material is not well known is to do hand measurements on the plate and then enter that data into a program, or do the entire analysis by hand. We have developed a small software package called CRYSTAL, for a PC/AT which is an intermediate point between the fully automated approach and the approach by hand. These programs were written to be used in conjunction with the EMS programs in order to compare theoretical patterns with those obtained experimentally.The program menu is displayed by typing “CRYSTAL”. There are five choices available from the menu: 1) enter diffraction pattern data using the digitizer, 2) analysis of the zone and planes associated with the measured planes, 3) plot the theoretical diffraction pattern and/or the measured planes, either on the CRT or the plotter, 4) calculate x-ray planes and 20 angles, 5) return to DOS.


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