scholarly journals Single-particle cryo-EM: alternative schemes to improve dose efficiency

2021 ◽  
Vol 28 (5) ◽  
pp. 1343-1356
Author(s):  
Yue Zhang ◽  
Peng-Han Lu ◽  
Enzo Rotunno ◽  
Filippo Troiani ◽  
J. Paul van Schayck ◽  
...  

Imaging of biomolecules by ionizing radiation, such as electrons, causes radiation damage which introduces structural and compositional changes of the specimen. The total number of high-energy electrons per surface area that can be used for imaging in cryogenic electron microscopy (cryo-EM) is severely restricted due to radiation damage, resulting in low signal-to-noise ratios (SNR). High resolution details are dampened by the transfer function of the microscope and detector, and are the first to be lost as radiation damage alters the individual molecules which are presumed to be identical during averaging. As a consequence, radiation damage puts a limit on the particle size and sample heterogeneity with which electron microscopy (EM) can deal. Since a transmission EM (TEM) image is formed from the scattering process of the electron by the specimen interaction potential, radiation damage is inevitable. However, we can aim to maximize the information transfer for a given dose and increase the SNR by finding alternatives to the conventional phase-contrast cryo-EM techniques. Here some alternative transmission electron microscopy techniques are reviewed, including phase plate, multi-pass transmission electron microscopy, off-axis holography, ptychography and a quantum sorter. Their prospects for providing more or complementary structural information within the limited lifetime of the sample are discussed.

Author(s):  
A. Ishikawa ◽  
C. Morita ◽  
M. Hibino ◽  
S. Maruse

One of the problems which are met in conventional transmission electron microscopy (CTEM) at high voltages is the reduction of the sensitivity of photographic films for high energy electron beams, resulting in the necessity of using high beam current. This cancels out an advantage of high voltage electron microscopy which is otherwise expected from the reduction of the inelastic scattering in the specimen, that is the reduced radiation damage of the specimen during observations. However, it is expected that the efficiency of the detector of scanning transmission electron microscopy (STEM) can be superior to that of CTEM, since the divergence of the electron beam in the detecting material does not affect the quality of the image. In addition to observation with less radiation damage, high voltage STEM with high detection efficiency is very attractive for observations of weak contrast objects since the enhancement of the contrast (which is an important advantage of STEM) is easily realized electrically.


2014 ◽  
Vol 20 (1) ◽  
pp. 175-183 ◽  
Author(s):  
Nicole Frindt ◽  
Marco Oster ◽  
Simon Hettler ◽  
Björn Gamm ◽  
Levin Dieterle ◽  
...  

AbstractTransmission electron microscopy (TEM) images of beam sensitive weak-phase objects such as biological cryo samples usually show a very low signal-to-noise ratio. These samples have almost no amplitude contrast and instead structural information is mainly encoded in the phase contrast. To increase the sample contrast in the image, especially for low spatial frequencies, the use of phase plates for close to focus phase contrast enhancement in TEM has long been discussed. Electrostatic phase plates are favorable in particular, as their tunable potential will allow an optimal phase shift adjustment and higher resolution than film phase plates as they avoid additional scattering events in matter. Here we show the first realization of close to focus phase contrast images of actin filament cryo samples acquired using an electrostatic Zach phase plate. Both positive and negative phase contrast is shown, which is obtained by applying appropriate potentials to the phase plate. The dependence of phase contrast improvement on sample orientation with respect to the phase plate is demonstrated and single-sideband artifacts are discussed. Additionally, possibilities to reduce contamination and charging effects of the phase plate are shown.


Author(s):  
A. Ourmazd

High Resolution Transmission Electron Microscopy (HRTEM) is now a powerful probe for the structural analysis of semiconductor systems. Lattice images can be obtained in a number of orientations, in at least three of which the individual atomic columns can be resolved. However, there exits an important class of problems, whose resolution requires chemical as well as structural information. The identification of individual atomic columns in compound semiconductors, and the atomic configuration of semiconductor/semiconductor interfaces are two important examples.In general, most reflection used to form a lattice image are not particularly sensitive to chemical changes in the sample. The information content of a typical lattice image is therefore strongly dominated by structural details. On the other hand, reflections such as the (200), which are normally forbidden in the diamond structure, come about in the zinc-blende system because of the chemical differences between the occupants of the two sublattices, and are thus highly chemically sensitive. In the “kinematical” thickness region, where simple image interpretation is possible, such reflections are relatively weak and their contribution to the lattice image is dominated by the stronger and chemically insensitive, allowed reflections.


Author(s):  
Robert C. Rau ◽  
John Moteff

Transmission electron microscopy has been used to study the thermal annealing of radiation induced defect clusters in polycrystalline tungsten. Specimens were taken from cylindrical tensile bars which had been irradiated to a fast (E > 1 MeV) neutron fluence of 4.2 × 1019 n/cm2 at 70°C, annealed for one hour at various temperatures in argon, and tensile tested at 240°C in helium. Foils from both the unstressed button heads and the reduced areas near the fracture were examined.Figure 1 shows typical microstructures in button head foils. In the unannealed condition, Fig. 1(a), a dispersion of fine dot clusters was present. Annealing at 435°C, Fig. 1(b), produced an apparent slight decrease in cluster concentration, but annealing at 740°C, Fig. 1(C), resulted in a noticeable densification of the clusters. Finally, annealing at 900°C and 1040°C, Figs. 1(d) and (e), caused a definite decrease in cluster concentration and led to the formation of resolvable dislocation loops.


Author(s):  
Joseph J. Comer ◽  
Charles Bergeron ◽  
Lester F. Lowe

Using a Van De Graaff Accelerator thinned specimens were subjected to bombardment by 3 MeV N+ ions to fluences ranging from 4x1013 to 2x1016 ions/cm2. They were then examined by transmission electron microscopy and reflection electron diffraction using a 100 KV electron beam.At the lowest fluence of 4x1013 ions/cm2 diffraction patterns of the specimens contained Kikuchi lines which appeared somewhat broader and more diffuse than those obtained on unirradiated material. No damage could be detected by transmission electron microscopy in unannealed specimens. However, Dauphiné twinning was particularly pronounced after heating to 665°C for one hour and cooling to room temperature. The twins, seen in Fig. 1, were often less than .25 μm in size, smaller than those formed in unirradiated material and present in greater number. The results are in agreement with earlier observations on the effect of electron beam damage on Dauphiné twinning.


Author(s):  
D. Loretto ◽  
J. M. Gibson ◽  
S. M. Yalisove

The silicides CoSi2 and NiSi2 are both metallic with the fee flourite structure and lattice constants which are close to silicon (1.2% and 0.6% smaller at room temperature respectively) Consequently epitaxial cobalt and nickel disilicide can be grown on silicon. If these layers are formed by ultra high vacuum (UHV) deposition (also known as molecular beam epitaxy or MBE) their thickness can be controlled to within a few monolayers. Such ultrathin metal/silicon systems have many potential applications: for example electronic devices based on ballistic transport. They also provide a model system to study the properties of heterointerfaces. In this work we will discuss results obtained using in situ and ex situ transmission electron microscopy (TEM).In situ TEM is suited to the study of MBE growth for several reasons. It offers high spatial resolution and the ability to penetrate many monolayers of material. This is in contrast to the techniques which are usually employed for in situ measurements in MBE, for example low energy electron diffraction (LEED) and reflection high energy electron diffraction (RHEED), which are both sensitive to only a few monolayers at the surface.


Author(s):  
L. Hultman ◽  
C.-H. Choi ◽  
R. Kaspi ◽  
R. Ai ◽  
S.A. Barnett

III-V semiconductor films nucleate by the Stranski-Krastanov (SK) mechanism on Si substrates. Many of the extended defects present in the films are believed to result from the island formation and coalescence stage of SK growth. We have recently shown that low (-30 eV) energy, high flux (4 ions per deposited atom), Ar ion irradiation during nucleation of III-V semiconductors on Si substrates prolongs the 1ayer-by-layer stage of SK nucleation, leading to a decrease in extended defect densities. Furthermore, the epitaxial temperature was reduced by >100°C due to ion irradiation. The effect of ion bombardment on the nucleation mechanism was explained as being due to ion-induced dissociation of three-dimensional islands and ion-enhanced surface diffusion.For the case of InAs grown at 380°C on Si(100) (11% lattice mismatch), where island formation is expected after ≤ 1 monolayer (ML) during molecular beam epitaxy (MBE), in-situ reflection high-energy electron diffraction (RHEED) showed that 28 eV Ar ion irradiation prolonged the layer-by-layer stage of SK nucleation up to 10 ML. Otherion energies maintained layer-by-layer growth to lesser thicknesses. The ion-induced change in nucleation mechanism resulted in smoother surfaces and improved the crystalline perfection of thicker films as shown by transmission electron microscopy and X-ray rocking curve studies.


2021 ◽  
Vol 27 (S1) ◽  
pp. 3358-3359
Author(s):  
Hyeokmin Choe ◽  
Eric Montgomery ◽  
Ilya Ponomarev ◽  
June Lau ◽  
Yimei Zhu ◽  
...  

2012 ◽  
Vol 18 (S2) ◽  
pp. 1138-1139
Author(s):  
Y. Inayoshi ◽  
H. Minoda

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.


1995 ◽  
Vol 401 ◽  
Author(s):  
L. Ryen ◽  
E. Olssoni ◽  
L. D. Madsen ◽  
C. N. L. Johnson ◽  
X. Wang ◽  
...  

AbstractEpitaxial single layer (001) SrTiO3 films and an epitaxial Yba2Cu3O7-x/SrTiO3 multilayer were dc and rf sputtered on (110)rhombohedral LaAIO3 substrates. The microstructure of the films was characterised using transmission electron microscopy. The single layer SrTiO3 films exhibited different columnar morphologies. The column boundaries were due to the lattice mismatch between film and substrate. The boundaries were associated with interfacial dislocations at the film/substrate interface, where the dislocations relaxed the strain in the a, b plane. The columns consisted of individual subgrains. These subgrains were misoriented with respect to each other, with different in-plane orientations and different tilts of the (001) planes. The subgrain boundaries were antiphase or tilt boundaries.The individual layers of the Yba2Cu3O7-x/SrTiO3 multilayer were relatively uniform. A distortion of the SrTiO3 unit cell of 0.9% in the ‘001’ direction and a Sr/Ti ratio of 0.62±0.04 was observed, both in correspondence with the single layer SrTiO3 films. Areas with different tilt of the (001)-planes were also present, within each individual SrTiO3 layer.


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