Cryoelectron microscopy and image reconstruction of spherical viruses with spot scan and FEG technologies

1995 ◽  
Vol 53 ◽  
pp. 1086-1087
Author(s):  
N. H. Olson ◽  
U. Lücken ◽  
S. B. Walker ◽  
M. T. Otten ◽  
T. S. Baker
Keyword(s):  
Electron Microscope ◽  
Transfer Function ◽  
Field Emission ◽  
Phase Contrast ◽  
Atomic Resolution ◽  
Bright Spot ◽  
Biological Macromolecules ◽  
Contrast Transfer Function ◽  
Transmission Electron ◽  
Chlorotic Mottle

The field emission gun electron microscope (FEG) is a tool that has the potential to achieve near atomic resolution information of biological macromolecules. The FEG provides a beam with higher spatial and temporal coherence and a better phase contrast transfer function than do microscopes with either tungsten or LaB6 filaments. The FEG is also ideal for spot scan imaging applications because it can produce a small, coherent and very bright spot. In spot scan mode the specimen is exposed to an array of nonoverlapping spots rather man a flood beam. This significantly reduces beam-induced specimen drift.Frozen-hydrated samples of cowpea chlorotic mottle (CCMV, Fig. 1A) and cowpea severe mosaic virus (CPSMV, Fig. IB) were examined on a Philips CM12 transmission electron microscope equipped with a standard LaB6 gun and on a Philips CM200 equipped with a field emission gun, respectively. The CM12 was operated at 120kV and was externally controlled by means of a spot scan imaging program which produced a series of 250 nm diameter spots on Kodak SO-163 sheet film.

The information limit of a 200kv field emission TEM

1995 ◽  
Vol 53 ◽  
pp. 586-587
Author(s):  
T. Kaneyama ◽  
M. Kawasaki ◽  
T. Tomita ◽  
T. Honda ◽  
M. Kersker
Keyword(s):  
Thin Film ◽  
Electron Microscope ◽  
Field Emission ◽  
Phase Contrast ◽  
Mechanical Stability ◽  
Power Supplies ◽  
Objective Lens ◽  
Operating Parameters ◽  
Contrast Transfer Function ◽  
Transmission Electron

The Point resolution of a transmission electron microscope is normally defined by the reciprocal of the spatial frequency of the first zero in the phase contrast transfer function at the Scherzer defocus condition. When a field emission gun (FEG) is used as the electron source, the information limit, that point at which the contrast beyond the first zero goes to zero contrast, becomes equally important. We have investigated the primary microscope parameters that affect the information limit.A 200kV FE-TEM (JEM-2010F) equipped with a ZrO/W shottkey emitter and Gatan Parallel EELS (PEELS) was used for the experiments. The aberration coefficients of the objective lens are Cs = 1mm and Cc = 1.4mm. The specimen used is an evaporated amorphous Ge thin film with small gold islands.The resolution performance of the microscope depends not only on the performance of the objective lens, the high voltage stability, stability of the lens and deflector power supplies, operating parameters of the FEG, and the overall mechanical stability of the microscopes.

scholarly journals Contrast Transfer Function-Based Exit-Wave Reconstruction and Denoising of Atomic-Resolution Transmission Electron Microscopy Images of Graphene and Cu Single Atom Substitutions by Deep Learning Framework

Nanomaterials ◽  
2020 ◽  
Vol 10 (10) ◽  
pp. 1977
Author(s):  
Jongyeong Lee ◽  
Yeongdong Lee ◽  
Jaemin Kim ◽  
Zonghoon Lee
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Transfer Function ◽  
Signal To Noise Ratio ◽  
Atomic Resolution ◽  
Single Atom ◽  
Monolayer Graphene ◽  
Signal To Noise ◽  
Contrast Transfer Function ◽  
Transmission Electron

The exit wave is the state of a uniform plane incident electron wave exiting immediately after passing through a specimen and before the atomic-resolution transmission electron microscopy (ARTEM) image is modified by the aberration of the optical system and the incoherence effect of the electron. Although exit-wave reconstruction has been developed to prevent the misinterpretation of ARTEM images, there have been limitations in the use of conventional exit-wave reconstruction in ARTEM studies of the structure and dynamics of two-dimensional materials. In this study, we propose a framework that consists of the convolutional dual-decoder autoencoder to reconstruct the exit wave and denoise ARTEM images. We calculated the contrast transfer function (CTF) for real ARTEM and assigned the output of each decoder to the CTF as the amplitude and phase of the exit wave. We present exit-wave reconstruction experiments with ARTEM images of monolayer graphene and compare the findings with those of a simulated exit wave. Cu single atom substitution in monolayer graphene was, for the first time, directly identified through exit-wave reconstruction experiments. Our exit-wave reconstruction experiments show that the performance of the denoising task is improved when compared to the Wiener filter in terms of the signal-to-noise ratio, peak signal-to-noise ratio, and structural similarity index map metrics.

Holographic contrast transfer theory

1992 ◽  
Vol 50 (2) ◽  
pp. 984-985
Author(s):  
R. Plass ◽  
L. D. Marks
Keyword(s):  
Electron Beam ◽  
Transfer Function ◽  
Field Emission ◽  
Electron Holography ◽  
Contrast Transfer Function ◽  
Beam Focusing ◽  
Emission Electron ◽  
Transmission Electron ◽  
Electron Microscopes ◽  
Image Position

With the advent of reliable cold field emission transmission electron microscopes there is substantial interest in using the amplitude and phase information recorded in electron holograms to optically or numerically correct for the coherent aberrations of transmission electron microscopes. However electron holography cannot compensate for incoherent aberrations. The derivation of the contrast transfer function for off axis electron holography in this paper shows there is no fundamental improvement in resolution for electron holography over conventional transmission electron microscopy.Evaluating the contrast transfer function involves mathematically following an electron beam through a field emission electron microscope set up for off axis electron holography. Due to the high coherence of the field emission electron beam coherent aberrations caused by the pre-specimen beam focusing system must be accounted for. Starting with a spacial frequency distribution, C(v), for the electron beam leaving the gun, the electron beam is limited by the condenser aperture and coherently aberrated by the condenser lens and objective pre-field as it passes to the specimen region:

Practical aspects about hollow-cone imaging

1995 ◽  
Vol 53 ◽  
pp. 576-577
Author(s):  
T. Geipel ◽  
W. Mader
Keyword(s):  
Electron Microscope ◽  
Transfer Function ◽  
Phase Contrast ◽  
Hollow Cone ◽  
Low Resolution ◽  
Contrast Transfer Function ◽  
Imaging Parameters ◽  
Specimen Holder ◽  
Optimum Imaging

Hollow-cone imaging (HCI) as a possibility to improve the resolution of a TEM has already been proposed in the late 40ties and besides others, there have been extensive hollow-cone experiments 10 years back using a low resolution TEM with a non-tilt specimen holder. In a recent paper the optimum imaging parameters for HCI were determined leading to an improvement of the resolution by a factor of two. However, there are contrast limitations and experimental problems for HCI which were only partly considered in Ref. and which will be discussed in this paper for a modern electron microscope. Preliminary experiments were performed which are not shown in the abstract.In Fig. 1 ∫ ct f(u)du is plotted versus defocus Δ f for different cone radii Θc and a fixed aperture radius Θo = 1/δ = 5 nm-1 (ct f is the phase contrast transfer function (PCTF) for HCI and δ = 0.2 nm is the resolution of a CM30 supertwin microscope).

Correlation of cryo-electron microscopic and x-ray data and compensation of the contrast transfer function

1992 ◽  
Vol 50 (2) ◽  
pp. 996-997
Author(s):  
R. Holland Cheng
Keyword(s):  
Transfer Function ◽  
Phase Contrast ◽  
Objective Lens ◽  
Biological Macromolecules ◽  
Electron Microscopic ◽  
Contrast Transfer Function ◽  
Image Reconstruction Techniques ◽  
X Ray ◽  
Mass Distributions ◽  
Spatial Frequencies

Cryo-electron microscopy (cryoEM) along with image reconstruction techniques can produce vivid images of biological macromolecules in their “native” state, although objective interpretation of these images is influenced by the fact that the contribution of phase contrast greatly exceeds that of amplitude contrast in such weakly scattering objects. The microscope contrast transfer function (CTF), which is strongly dependent on the defocus level of objective lens, modulates images of the object density distribution as a function of spatial frequency. Compensation for the effects of phase contrast transfer is important because underweighting of the low spatial frequencies usually causes difficulties in evaluating absolute mass distributions in objects.Correct compensation for the CTF is difficult to achieve. This is due, in part, to ambiguities in measuring the exact defocus level in noisy micrographs, and in knowing the relative contributions of amplitude and phase contrast, beam coherence, and inelastic scattering. The availability of atomic resolution determinations for a few viruses allows one to determine empirically how to correct the cryoEM images to best fit the x-ray data.

Observation of biological macromolecules using various electron microscopic methods

1978 ◽  
Vol 36 (2) ◽  
pp. 170-171
Author(s):  
Mitsuo Ohtsuki ◽  
Michael Sogard
Keyword(s):  
Electron Microscope ◽  
Phase Contrast ◽  
Image Interpretation ◽  
Density Lipoprotein ◽  
Biological Macromolecules ◽  
Contrast Effects ◽  
Biological Structure ◽  
Electron Microscopic ◽  
Structural Investigations ◽  
Staining Techniques

Structural investigations of biological macromolecules commonly employ CTEM with negative staining techniques. Difficulties in valid image interpretation arise, however, due to problems such as variability in thickness and degree of penetration of the staining agent, noise from the supporting film, and artifacts from defocus phase contrast effects. In order to determine the effects of these variables on biological structure, as seen by the electron microscope, negative stained macromolecules of high density lipoprotein-3 (HDL3) from human serum were analyzed with both CTEM and STEM, and results were then compared with CTEM micrographs of freeze-etched HDL3. In addition, we altered the structure of this molecule by digesting away its phospholipid component with phospholipase A2 and look for consistent changes in structure.

A Field Emission System For Transmission Electron Microscopy

1973 ◽  
Vol 31 ◽  
pp. 298-299
Author(s):  
Michel Troyonal ◽  
Huei Pei Kuoal ◽  
Benjamin M. Siegelal
Keyword(s):  
Electron Microscope ◽  
Field Emission ◽  
Optical System ◽  
High Vacuum ◽  
Ultra High Vacuum ◽  
Objective Lens ◽  
Electron Optical System ◽  
Cross Sectional ◽  
Transmission Electron ◽  
Emission System

A field emission system for our experimental ultra high vacuum electron microscope has been designed, constructed and tested. The electron optical system is based on the prototype whose performance has already been reported. A cross-sectional schematic illustrating the field emission source, preaccelerator lens and accelerator is given in Fig. 1. This field emission system is designed to be used with an electron microscope operated at 100-150kV in the conventional transmission mode. The electron optical system used to control the imaging of the field emission beam on the specimen consists of a weak condenser lens and the pre-field of a strong objective lens. The pre-accelerator lens is an einzel lens and is operated together with the accelerator in the constant angular magnification mode (CAM).

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