scholarly journals Spectra optimizes the use of electron dose

2021 ◽  
Vol 27 (S1) ◽  
pp. 1066-1067
Author(s):  
Eric Van Cappellen ◽  
Christian Maunders ◽  
Ingrid Kieft ◽  
Maarten Bischoff ◽  
Felix Van Uden ◽  
...  
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):  
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):  
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.


Author(s):  
R.D. Leapman ◽  
S.B. Andrews

Elemental mapping of biological specimens by electron energy loss spectroscopy (EELS) can be carried out both in the scanning transmission electron microscope (STEM), and in the energy-filtering transmission electron microscope (EFTEM). Choosing between these two approaches is complicated by the variety of specimens that are encountered (e.g., cells or macromolecules; cryosections, plastic sections or thin films) and by the range of elemental concentrations that occur (from a few percent down to a few parts per million). Our aim here is to consider the strengths of each technique for determining elemental distributions in these different types of specimen.On one hand, it is desirable to collect a parallel EELS spectrum at each point in the specimen using the ‘spectrum-imaging’ technique in the STEM. This minimizes the electron dose and retains as much quantitative information as possible about the inelastic scattering processes in the specimen. On the other hand, collection times in the STEM are often limited by the detector read-out and by available probe current. For example, a 256 x 256 pixel image in the STEM takes at least 30 minutes to acquire with read-out time of 25 ms. The EFTEM is able to collect parallel image data using slow-scan CCD array detectors from as many as 1024 x 1024 pixels with integration times of a few seconds. Furthermore, the EFTEM has an available beam current in the µA range compared with just a few nA in the STEM. Indeed, for some applications this can result in a factor of ~100 shorter acquisition time for the EFTEM relative to the STEM. However, the EFTEM provides much less spectral information, so that the technique of choice ultimately depends on requirements for processing the spectrum at each pixel (viz., isolated edges vs. overlapping edges, uniform thickness vs. non-uniform thickness, molar vs. millimolar concentrations).


Author(s):  
Zhifeng Shao ◽  
Ruoya Ho ◽  
Andrew P. Somlyo

Electron energy loss spectroscopy (EELS) has been a powerful tool for high resolution studies of elemental distribution, as well as electronic structure, in thin samples. Its foundation for biological research has been laid out nearly two decades ago, and in the subsequent years it has been subjected to rigorous, but by no means extensive research. In particular, some problems unique to EELS of biological samples, have not been fully resolved. In this article we present a brief summary of recent methodological developments, related to biological applications of EELS, in our laboratory. The main purpose of this work was to maximize the signal to noise ratio (S/N) for trace elemental analysis at a minimum dose, in order to reduce the electron dose and/or time required for the acquisition of high resolution elemental maps of radiation sensitive biological materials.Based on the simple assumption of Poisson distribution of independently scattered electrons, it had been generally assumed that the optimum specimen thickness, at which the S/N is a maximum, must be the total inelastic mean free path of the beam electron in the sample.


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