bluuebunny/crossref_metadata_2025_split huggingface dataset:
High Pressure Freezing has Come of Age - but is it Mature?
Authors: McDonald Kent 
Published: July 2020

Abstract
It should be no secret by now that ultrarapid freezing is a superior method of specimen preparation for many biological EM projects, and that high pressure freezing is the most versatile of the freezing methods. While cryopreservation is not necessary for all EM studies, it is the method of choice for high resolution work and where “fixation artifacts”, such as distorted membranes, or extraction of the cytosol is a problem. It is true that the machines are expensive, and not always immediately available, but for some questions it is the appropriate technology to use. In the U.S., there are machines available for general use in the far West (Berkeley, CA), the upper Midwest (Madision, WI and Minneapolis, MN), and in the Northeast (Albany, NY). For locations of any of the other 8 machines around the country, interested users can call Technotrade, International at (603) 622-5011.
Current Status of High Pressure Freezing: A decade ago, Studer et al. wrote an article entitled: “High Pressure Freezing Comes of Age” that illustrated how high pressure freezing (HPF) had become a proven technology, useful for preserving ultrastructure with unmatched fidelity in cell types that had previously been difficult to fix well for electron microscopy (EM).
Using High Pressure Freezing and Freeze Substitution to Investigate Kinetochore Ultrastructure in Vetebrate Somatic Cells
Authors: McEwen BF , Hsieh CE , Barnard RM , Rieder C L 
Published: July 2020

Abstract
In vertebrate somatic cells, sister kinetochores attach chromosomes to the mitotic spindle by capturing microtubules (Mts) emanating from opposite spindle poles. Kinetochores are also required for chromosome alignment at the spindle equator, stabilizing kinetochore Mts, cell cycle control of anaphase onset, and poleward migration of sister chromatids during anaphase. Thus kinetochores play a critical role in the distribution of genetic information to daughter cells during cell division. Understanding the molecular mechanisms behind kinetochore function requires knowing how the molecular components are arranged relative to each other, and relative to the kinetochore Mts. Currently, however, our knowledge of kinetochore composition is incomplete, and until recently our knowledge of kinetochore ultrastructure was limited to the familiar trilamellar model derived from conventionally fixed and dehydrated specimens(Figures 1a&b).
Highly fibrous structures, such as the kinetochore, are particularly vulnerable to distortions caused by the chemical fixation and dehydration methods used in conventional specimen preparations. Such distortions can be largely avoided by using high pressure freezing and freeze substitution (HPF/FS).
Combined Cy3 / Nanogold Conjugates for ImmunocytoChemistry and in Situ Hybridization
Authors: Powell Richard D , Joshi Vishwas N , Halsey Carol M R , Hainfeld James F , Hacker Gerhard W , Hauser-Kronberger Cornelia , Muss Wolfgang H , Takvorian Peter M 
Published: July 2020

Abstract
Fluorescein and the 1.4 nm Nanogold® gold cluster label may be incorporated into a single Fab’ immunoprobe by separate cross-linking reactions, to give a probe which labels antigenic sites in a single step for correlative fluorescence and electron microscope visualization. These probes show high labeling density, labeling a pre-mRNA splicing factor in the HeLa cell nucleus; Microtubules were also densely labeled using fluorescence, other optical modalities, and electron microscopy; in a parallel experiment, a 5 nm colloidal gold probe gave only occasional labeling. We now describe Fab’ and streptavidin probes containing both Nanogold® and the fluorescent cyanine dye, Cy3.
F(ab’)2 Goat anti-Mouse IgG and F(ab’)2 goat anti-rabbit IgG fragments were reductively cleaved to Fab’ fragments using dithiothreitol (DTT) or mercaptoethylamine hydrochloride (MEA), which selectively reduce the F(ab’)2 hinge disulfide bonds, with 5 mm EDTA to prevent reoxidation. Fab’ fragments were isolated by gel filtration (coarse gel: GH25, Amicon) then labeled with Monomaleimido- Nanogold® which reacts site-specifically with thiols. Streptavidin was labeled using Mono- Sulfo-NHS-Nanogold® at pH 7.5. Nanogold® conjugates were isolated by gel filtration (Superose-12 column, Pharmacia), then reacted with excess Cy3 monofunctional NHS ester (labeling kit, Amersham Life Sciences) at pH 7.5; dual-labeled conjugates were isolated by gel filtration (Superose-12).
Non-Imaging Microscopies: Flow Cytometry as a Correlative Analytical Tool in the Quantification of Cell Structure, Autofluorescence, Fluorescent Probes and Cell Populations.
Authors: Smith R W 
Published: July 2020

Abstract
Non-imaging microscopy has developed somewhat independently of both traditional light microscopy and laser confocal microscopy. Flow cytometry is the chief commercial and research technology among these microscopies, and, like other nonimaging detection systems, developed around the theme of automation in clinical laboratory medicine. It is an important correlative or parallel microscopy to several image forming microscopical methods. Cell sorting is an important option as well.
The basic structure of the flow cytometer certainly parallels light, laser and electron microscopes. The flow cytometer has a light source, a set of adjustable optics to focus the beam on the specimen, objective optics to collect the light and direct it to appropriate sensors, and the sensors themselves. A real image is not formed because the sensors are not in an even plane with the projection, such as provided by the retina in light microscopy or an image plane or film plate in electron microscopy, and the objective optics may not focus in the image plane.
While early flow cytometers were developed primarily for the automatic counting of cells and particles, modern instruments offer particular advantages for the analysis of fluorescence, fluorescent chemicals and probes and cellular auto fluorescence.
Immunohistochemical and Morphometric Characterization of Blood Vessels in Brain Metastasis in Nude Mice
Authors: Bucana C D , Yano S , Reynolds D , Dunner K , Fidler I J 
Published: July 2020

Abstract
This study was conducted to compare the immunohistochemical and morphometric characteristics of blood vessels in brain metastasis with blood vessels of the adjacent uninvolved brain tissue. This is based on the hypothesis that new vessels formed in the tumor express antigens that are not expressed in blood vessels of the adult brain parenchyma. Thus, a panel of antibodies known to react with mouse endothelial cells at various stages of differentiation or stimulation was tested on frozen sections of experimental brain metastasis. Intratumoral microvessel density which has been reported to reflect the intensity of tumor angiogenesis was inapplicable to experimental brain metastasis therefore other parameters were measured.
Brain metastasis was produced in nude mice following intracarotid injection of tumor cells. Three human tumors and three murine tumors were injected into the carotid artery of nude mice. The animals were sacrificed when they were moribund, the brains were embedded in OCT and frozen in liquid nitrogen. Some animals were injected with bromodeoxyuridine (BrdU) 1 h before they were necropsied. Immunohistochemistry was performed using antibodies directed to the following antigens: CD31, CD34, MECA-32, endoglin, ICAM-1, ICAM-2, and BrdU. Antibodies to leukocyte antigens, F4/80 and CD3 were also tested to determine the amount of host cells infiltrating the tumors. SEM of immunogold labeled preparations determined the cellular localization of some antigens.
A Compilation of Biological SEM Sample Prepartion Techniques for Application to Outreach Programs
Authors: Smith NR , Duarte E , Morgan C , Quinta RA 
Published: July 2020

Abstract
A partnership between California State University, Hayward and Ohlone College, Fremont, California resulted in a program to introduce undergraduate students to scanning electron microscopy (SEM) sample preparation procedures. Ten students enrolled in a biology major's course at the Ohlone Community College are selected each semester to participate in the program. In addition to these students, ten others from a Basic Skills Enhancement program for underrepresented students participate through a short course offered during the summer. The program, in effect for two years, resulted in the compilation of a laboratory manual describing basic sample preparation techniques that are appropriate for an outreach program. We select samples that are simple to prepare yet yield results that are germane to a biology student's curriculum. A selection of plants, pollen grains, bacteria, fungi, yeast, Drosophilia melanogaster, and various other invertebrates are typical samples that lend themselves to an outreach program that emphasizes biological concepts. Initially our program required the Ohlone College students to come to the Hayward campus to prepare their samples. External funding has allowed us to purchase sample preparation equipment for the community college.
An Introduction Electronic Documentation Preparation and Submission for Future Microscopy and Microanalysis Meetings.
Authors: Mansfield John F 
Published: July 2020

Abstract
Since the introduction of personal computers in the early 1980s, most documents and manuscripts have been prepared from within a word processing program on such a computer. Indeed, most documents on personal computers are created on a limited number of word processing packages that have come to dominate the market. Most of the data acquisition that attendees of Microscopy and Microanalysis currently perform is digital, i.e. directly into a personal computer or desktop workstation. It is frequently the case that images, spectra and diffraction patterns are only committed to paper when they are dispatched for publication. With the advent of the World Wide Web in 1994, and its subsequent explosive growth, entirely electronic publishing has become possible. It is possible to envisage the day when most publications will be available solely electronically and material will be only printed out occasionally by the reader.
While there are a growing number of journals that are only available electronically, entirely paperless publication is not yet the norm and there are no definite plans, as yet, to publish the proceedings of the Microscopy and Microanalysis meetings solely in electronic format. However, there are a number of advantages to applying some of these emerging technologies in the production of the future proceedings. Electronic submission of abstracts is one advance that is being considered by the organizers of the meetings scheduled for the early 2000s. The National Science Foundation now receives almost all proposals electronically. There are several reasons for moving to electronic submission. The first is that proofing and correcting the abstracts would then be straightforward.
Development of Electron Probe Instrumentation During those Early Days When Professor Castatng Visited Japan
Authors: Shimizu R 
Published: July 2020

Abstract
In May of 1955 Professor Shinoda introduced the development of the electron probe microanalyzer (EPMA) in a review article published by Oyobutsuri (the Applied Physics journal written in Japanese), which showed a schematic diagram of an EPMA system. That paper, published in the most popular journal in Japan, was very probably the first paper which drew the attention of the people involved in the field of material science, in particular, the field of metallurgy, who were especially interested in EPMA. That paper stimulated the development of EPMA in Japan.
In 1957, the funding provided by the Grant-in-Aid for Developmental Research to the university research groups headed by Profs. Akutagawa, Gokyuu, and Abe at the University of Tokyo was used, in a cooperation with the Hitachi group headed by Drs. Tadano and Watanabe, to develop the EPMA in Japan. Their research results led to the funding of a 2 year Grant-in-Aid for Cooperative Research project (April 1960 to March 1962) called “The Study of X-ray Microanalyzer”, which was chaired by Prof. Sakaki and consisted of 37 members from various universities, national institutes and industries. Prof. Shinoda, who became the Chairman of that project in April of 1962, led that group for another year until March of 1963.
It was just after the start of the Cooperative Research project, in September of 1960, that Professor Castaing visited Japan and gave several talks as a representative of the French Mission Culturelle. Professor Castaing gave a talk at the Hitachi Central Research Laboratory on the 19th of September, another talk at the Nagoya University on the 24th of September, and also a talk at Osaka University on the 28th of September. During his stay in Japan from September 18th to October 4th he talked about his work on EPMA a total of six times by also giving two informal presentations. The visit of Professor Castaing gave a great push forward for the commercial development of EPMA instruments in Japan.
Application of the Electron Probe Microanalyzer to the Analysis of Planetary and Geological Materials: a Historical Perspective.
Authors: Keil Klaus 
Published: July 2020

Abstract
Castaing first presented his idea to use secondary X-rays excited by a focused electron beam from a polished solid sample for microanalysis in 1949 at the First European Regional Conference on Electron Microscopy in Delft, Netherlands. As part of his dissertation, he then not only built the first electron probe microanalyzer (EPM), but also established many of the theoretical and analytical principles of the technique. This technique offered enormous analytical advantages to earth scientists over other analytical methods available at the time. For example, it allows qualitative and quantitative analysis of individual mineral grains a few microns in diameter; mineral grains can be viewed during analysis, thus ensuring accurate correlation between composition and grain morphology; for most purposes, the method is non-destructive; in situ analysis of minerals in polished thin sections results in retention of textural relationships among coexisting minerals; because of the small excitation volume, the technique is ideally suited for the study of zoned minerals, minute inclusions, exsolution lamellae, etc.; and once suitable standards are prepared and correction procedures are established, a large number of quantitative analyses can be obtained in a comparatively short time.
Analysis of Metallic Pigments Used in 19th Century Japanese Prints
Authors: Hashimoto RY , Menon ESK , Fiorillo J 
Published: July 2020

Abstract
Japanese color woodblock prints ("ukiyo-e") from the Edo (1615-1868) and Meiji periods (1868- 1912) are among the most celebrated examples of color printmaking in the world. Full-color prints became technically and commercially viable around 1765. Traditional ukiyo-e colorants were watersoluble, uniform, translucent, organic (vegetal) and inorganic pigments of very small particle size. (Synthetic colorants were also introduced from the West in the 19th C, probably beginning with "Prussian blue" in the 1820s.) A pigment was applied with a brush to a color block, and then rice flour paste was mixed with the pigment to thicken it, facilitate control and rubbing into the paper, improve the uniformity of the colorant, and avoid a dry or granular texture. The dampened paper, sized with cooked bovine skins and alum, was then placed on the block (aligning with registration marks cut into the blocks). The colorants were then pressed into the paper by rubbing a bamboo pad against the back of the paper so that the colors were absorbed on the front surface.
SEM and EDS Characterization of Palladium Cathodes After Electrolysis in Light and Heavy Water
Authors: Silver D , Dash J 
Published: July 2020

Abstract
Studies of Pd foil cathodes have indicated that when electrolyzed for long times in electrolytes containing both deuterium and hydrogen ions along with sulfuric acid, the cathodes undergo compositional, topographical, evolutionary, and isotopic changes. The results reported here describe the changes which are produced in Pd cathodes after only a few minutes of electrolysis.
In June 1993 a cold-rolled, Pd (AESAR lot #12E06) cathode of 40μm thickness was submerged at one end into an electrolyte containing 20 ml D2O and 3.5 ml H2SO4 and electrolyzed for six minutes at about 0.6 A/cm2 and 3.65 V. An identical Pd cathode (cut from the same strip) was submerged at one end into an electrolyte containing 20 ml H2O and 3.5 H2SO4 and electrolyzed for six minutes using 0.6 A/cm2 and 4.11 V. A Pt foil acted as the anode in each case. The two cells were connected in series. An ionic current flowed between the electrodes for each system as the voltages were applied.
Simulation of Energy Deposition in E-Beam Irradiated Polymers
Authors: Horny Paula , Gauvin Raynald , Hovington Pierre , Besner Simon 
Published: July 2020

Abstract
Introduction
Radiation curing of polymers, by UV light or E-beam, is of growing interest in the state-of-the-art industry. Radiation curing is solvent free and leads to important energy and money savings. Improved materials characteristics, better control of doses and more accurate application allow radiation curing to take a big part of the surface coating industry and, for the last ten years, the polymeric and microelectronics (lithography) industry. But behaviors of polymers under radiant source is often empirically characterized. In this abstract, it is proposed a way to obtain accurate energy distribution from Monte Carlo simulations. For bulk or thin polymeric films, it is therefore essential to simulate energy distribution, in order to foresee what happens in the sample.
Results
CASINO is a single scattering Monte Carlo Simulation of electroN trajectory in sO lid -particularly designed for low energy beam interaction in a bulk and thin foil.
Trace Element Mapping of Minerals and Materials by Electron Microprobe
Authors: Fournelle J , Davidson C , Spear F , Kohn M , Guo H 
Published: July 2020

Abstract
A strength of the modern electron microprobe is its ability to provide 2D compositional information about materials. These images give the ability to observe features that might otherwise pass unseen. Elements at the trace element level are generally ignored due to the high detection limits imposed by mapping under “standard” EMP conditions.
Trace element mapping requires beam regulation at high (e.g. 300 nA) to very high (e.g. 3 μA) faraday cup currents, reliable beam and stage control, and suitable samples and mounting media. The ability to operate at high accelerating voltage to maximize Pk2/Bkg is desirable (Robinson and Graham, 1992), although we have encountered column difficulties above 25-30 kV.
We are mapping trace and minor elements including Y, Sc, P, Cr, Mn, Ca, in garnets. Fig. 1 shows Y, Sc and Cr maps (Spear and Kohn, 1996; Kohn, Spear and Valley, 1997), and Fig. 2 Y and Sc maps (Cameron, unpub. data), produced with a Cameca SX51.
Integration of a Monochromator in the Field Emission STEM
Authors: Mookb HW , Batsona PE 
Published: July 2020

Abstract
It would be very desirable to obtain EELS at 0. l-0.2eV resolution for bandstructure studies at interfaces and defects. [1,2] It has been possible to build spectrometers that perform in this energy range, but it has been difficult to utilize this resolution, because the energy width of the cold field emission source is about 0.3-0.4 eV for the currents needed to do practical microscopy. [3] Numerical methods have been used to sharpen the spectra, showing unambiguously that EELS spectra would benefit greatly if a higher resolution source were available. [4,5]
This situation has led one of us (H.W. Mook) to investigate electron monochromator optical designs which would be compatible with the high brightness and current required by the STEM. [6] This work produced a 4cm long monochromator that can be integrated into the field emission source. As shown in Fig. 1, the monochromator optics includes a field lens formed by deceleration of the electron beam from the 3 KeV extraction energy to about 350 eV inside the monochromator, a short Wien Filter velocity dispersive field, 150nm wide Si energy selecting slits, and an acceleration field lens at the monochromator exit.
The Electronic Band -Gap of Polycrystalline Diamond and Tetrahedral Amorphous Carbon by EELS in a STEM.
Authors: Papworth AJ , Kiely CJ , Amaratunga GAJ 
Published: July 2020

Abstract
Good quality polycrystalline diamond is transparent and has an optical band-gap very close to that of single crystal diamond, 5.5 eV. Tetrahedral amorphous carbon (ta-C) is a form of amorphous carbon which can have up to 90 % sp3 type bonds. It is the form of amorphous carbon, which is closest to having a true diamond-like structure. The optical band-gap of ta-C is however very different from that of polycrystalline diamond, typically in the range 2-3 eV. This difference in the optical bandgap has been ascribed to the electronic states formed by the residual graphitic bonding. The π − π* states at the graphitic sites effectively form tail states on the valance and conduction band edges created by the diamond-like bonding. These tail states extend deep into the band-gap, which would exist if there were only (100 %) diamond-like bonds. The poor electronic properties obtained thus far for ta-C are now accepted as being associated with localised conduction in the π − π* tail states.
Insights into the Electronic Structure of Ceramics Through Quantitative Analysis of Valence Electron Energy-Loss Spectroscopy (VEELS)
Authors: Müllejans Harald , H. French Roger 
Published: July 2020

Abstract
The electronic structure of ceramics can be extracted quantitatively from the valence electron energy-loss spectroscopy (VEELS) of transitions between the valence and conduction bands. We obtained VEEL spectra of several ceramics (FIG. 1) with a VG HB501 dedicated STEM equipped with Gatan PEELS. Improved data acquisition and new methods of data analysis allowed us to treat the data fully quantitatively. The reliable and accurate removal of the zero loss peak was crucial because intensities at energy losses just above the band gap of the ceramic material have a large influence on the results. An asymmetric Pearson VII function was fitted into the zero loss peak up to an energy loss for which no transitions are expected (an energy smaller than the band gap of the ceramic) and then extrapolated to higher energies. This limits the analysis to non-metallic materials, exhibiting non-zero band gap energies. We are currently developing methods to perform the analysis on metallic materials, using ellipsometric data in the visible and extrapolate the energy-loss function to 0 eV and thereby remove the need for the no transition energy.
Stem Without Spherical Aberration
Authors: Krivanek OL , Dellby N , Lupini AR 
Published: July 2020

Abstract
Even though two generations of electron microscopists have come to accept that the resolution of their instruments is limited by spherical aberration, three different aberration correctors showing that the aberration can be overcome have recently been built [1-3]. One of these correctors was developed by us specifically for forming small electron probes in a dedicated scanning transmission electron microscope (STEM) [3, 4]. It promises to revolutionize the way STEM instruments are built and the types of problems that they are applied to.
As was the case with the Berlin Wall, when a barrier that was once thought immovable finally crumbles, many of the consequences can be quite unexpected. For STEM, the removal of spherical aberration (Cs) as the main resolution limit is likely to lead to a new paradigm in which:
1) The resolution at a given operating voltage will improve by about 3x relative to today's best. When Cs can be adjusted arbitrarily in a STEM being used for microanalysis or dark field imaging, defocus and Cs are set to values that optimally oppose the effect of the 5th-order spherical aberration C5.