To Monochromate or not to Monochromate: Balancing Electron Dose and Energy Spread Requirements

2021 ◽  
Author(s):  
Frances Quigley
Keyword(s):  
Author(s):  
Kenneth H. Downing ◽  
Robert M. Glaeser

The structural damage of molecules irradiated by electrons is generally considered to occur in two steps. The direct result of inelastic scattering events is the disruption of covalent bonds. Following changes in bond structure, movement of the constituent atoms produces permanent distortions of the molecules. Since at least the second step should show a strong temperature dependence, it was to be expected that cooling a specimen should extend its lifetime in the electron beam. This result has been found in a large number of experiments, but the degree to which cooling the specimen enhances its resistance to radiation damage has been found to vary widely with specimen types.


Author(s):  
L. D. Jackel

Most production electron beam lithography systems can pattern minimum features a few tenths of a micron across. Linewidth in these systems is usually limited by the quality of the exposing beam and by electron scattering in the resist and substrate. By using a smaller spot along with exposure techniques that minimize scattering and its effects, laboratory e-beam lithography systems can now make features hundredths of a micron wide on standard substrate material. This talk will outline sane of these high- resolution e-beam lithography techniques.We first consider parameters of the exposure process that limit resolution in organic resists. For concreteness suppose that we have a “positive” resist in which exposing electrons break bonds in the resist molecules thus increasing the exposed resist's solubility in a developer. Ihe attainable resolution is obviously limited by the overall width of the exposing beam, but the spatial distribution of the beam intensity, the beam “profile” , also contributes to the resolution. Depending on the local electron dose, more or less resist bonds are broken resulting in slower or faster dissolution in the developer.


Author(s):  
T. A. Welton

Various authors have emphasized the spatial information resident in an electron micrograph taken with adequately coherent radiation. In view of the completion of at least one such instrument, this opportunity is taken to summarize the state of the art of processing such micrographs. We use the usual symbols for the aberration coefficients, and supplement these with £ and 6 for the transverse coherence length and the fractional energy spread respectively. He also assume a weak, biologically interesting sample, with principal interest lying in the molecular skeleton remaining after obvious hydrogen loss and other radiation damage has occurred.


Author(s):  
D.T. Grubb

Diffraction studies in polymeric and other beam sensitive materials may bring to mind the many experiments where diffracted intensity has been used as a measure of the electron dose required to destroy fine structure in the TEM. But this paper is concerned with a range of cases where the diffraction pattern itself contains the important information.In the first case, electron diffraction from paraffins, degraded polyethylene and polyethylene single crystals, all the samples are highly ordered, and their crystallographic structure is well known. The diffraction patterns fade on irradiation and may also change considerably in a-spacing, increasing the unit cell volume on irradiation. The effect is large and continuous far C94H190 paraffin and for PE, while for shorter chains to C 28H58 the change is less, levelling off at high dose, Fig.l. It is also found that the change in a-spacing increases at higher dose rates and at higher irradiation temperatures.


Author(s):  
H.A. Cohen ◽  
W. Chiu

The goal of imaging the finest detail possible in biological specimens leads to contradictory requirements for the choice of an electron dose. The dose should be as low as possible to minimize object damage, yet as high as possible to optimize image statistics. For specimens that are protected by low temperatures or for which the low resolution associated with negative stain is acceptable, the first condition may be partially relaxed, allowing the use of (for example) 6 to 10 e/Å2. However, this medium dose is marginal for obtaining the contrast transfer function (CTF) of the microscope, which is necessary to allow phase corrections to the image. We have explored two parameters that affect the CTF under medium dose conditions.Figure 1 displays the CTF for carbon (C, row 1) and triafol plus carbon (T+C, row 2). For any column, the images to which the CTF correspond were from a carbon covered hole (C) and the adjacent triafol plus carbon support film (T+C), both recorded on the same micrograph; therefore the imaging parameters of defocus, illumination angle, and electron statistics were identical.


Author(s):  
N. H. Olson ◽  
T. S. Baker ◽  
Wu Bo Mu ◽  
J. E. Johnson ◽  
D. A. Hendry

Nudaurelia capensis β virus (NβV) is an RNA virus of the South African Pine Emperor moth, Nudaurelia cytherea capensis (Lepidoptera: Saturniidae). The NβV capsid is a T = 4 icosahedron that contains 60T = 240 subunits of the coat protein (Mr = 61,000). A three-dimensional reconstruction of the NβV capsid was previously computed from visions embedded in negative stain suspended over holes in a carbon film. We have re-examined the three-dimensional structure of NβV, using cryo-microscopy to examine the native, unstained structure of the virion and to provide a initial phasing model for high-resolution x-ray crystallographic studiesNβV was purified and prepared for cryo-microscopy as described. Micrographs were recorded ∼1 - 2 μm underfocus at a magnification of 49,000X with a total electron dose of about 1800 e-/nm2.


Author(s):  
Max T. Otten ◽  
Wim M.J. Coene

High-resolution imaging with a LaB6 instrument is limited by the spatial and temporal coherence, with little contrast remaining beyond the point resolution. A Field Emission Gun (FEG) reduces the incidence angle by a factor 5 to 10 and the energy spread by 2 to 3. Since the incidence angle is the dominant limitation for LaB6 the FEG provides a major improvement in contrast transfer, reducing the information limit to roughly one half of the point resolution. The strong improvement, predicted from high-resolution theory, can be seen readily in diffractograms (Fig. 1) and high-resolution images (Fig. 2). Even if the information in the image is limited deliberately to the point resolution by using an objective aperture, the improved contrast transfer close to the point resolution (Fig. 1) is already worthwhile.


Author(s):  
T. Oikawa ◽  
N. Mori ◽  
T. Katoh ◽  
Y. Harada ◽  
J. Miyahara ◽  
...  

The “Imaging Plate”(IP) is a highly sensitive image recording plate for X-ray radiography. It has been ascertained that the IP has superior properties and high practicability as an image recording material in a TEM. The sensitivity, one of the properties, is about 3 orders higher than that of conventional photo film. The IP is expected to be applied to low dose techniques. In this paper, an estimation of the quantum noise on the TEM image which appears in case of low electron dose on the IP is reported.In this experiment, the JEM-2000FX TEM and an IP having the same size as photo film were used.Figure 1 shows the schematic diagram of the total system including the TEM used in this experiment. In the reader, He-Ne laser light is scanned across the IP, then blue light is emitted from the IP.


Author(s):  
Raynald Gauvin ◽  
Gilles L'Espérance

Values of cross sections for ionization of inner-shell electrons by electron impact are required for electron probe microanalysis, Auger-electron spectroscopy and electron energy-loss spectroscopy. In this work, the results of the measurement of inner-shell ionization cross-sections by electron impact, Q, in a TEM are presented for the K shell.The measurement of QNi has been performed at 120 KeV in a TEM by measuring the net X-ray intensity of the Kα line of Ni, INi, which is related to QNi by the relation :(1)where i is the total electron dose, (Ω/4π)is the fractional solid angle, ω is the fluorescence yield, α is the relative intensity factor, ε is the Si (Li) detector efficiency, A is the atomic weight, ρ is the sample density, No is Avogadro's number, t' is the distance traveled by the electrons in the specimen which is equal to τ sec θ neglecting beam broadening where τ is the specimen thickness and θ is the angle between the electron beam and the normal of the thin foil and CNi is the weight fraction of Ni.


Author(s):  
Neng-Yu Zhang ◽  
Terence Wagenknecht ◽  
Michael Radermacher ◽  
Tom Obrig ◽  
Joachim Frank

We have reconstructed the 40S ribosomal subunit at a resolution of 4 nm using the single-exposure pseudo-conical reconstruction method of Radermacher et al.Small (40S) ribosomal subunits were Isolated from rabbit reticulocytes, applied to grids and negatively stained (0.5% uranyl acetate) in a manner that “sandwiches” the specimen between two layers of carbon. Regions of the grid exhibiting uniform and thick staining were identified and photographed twice (magnification 49,000X). The first micrograph was always taken with the specimen tilted by 50° and the second was of the Identical area untilted (Fig. 1). For each of the micrographs the specimen was subjected to an electron dose of 2000-3000 el/nm2.Three hundred thirty particles appearing in the L view (defined in [4]) were selected from both tilted- and untilted-specimen micrographs. The untilted particles were aligned and their rotational alignment produced the azimuthal angles of the tilted particles in the conical tilt series.


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