bluuebunny/crossref_metadata_2025_split huggingface dataset:
Application of EM to differentiate herpes virus from pox virus in genital specimens
Authors: Rekrut K. , Schleuter K. 
Published: June 2020

Confirmation of herpes simplex virus (HSV) from genital lesions of obstetrical (OB) patients may affect both the management of the delivery and of the neonate.(l,2) During 1992 and 1993, 4,450 genital specimens from OB patients were submitted in viral transport media for herpes culture. The specimens were inoculated into MRC-5, Vero, and A-549 tissue culture tubes, incubated, and examined daily for 7 days for cytopathic effect (CPE). The original specimens were frozen at −70° C until final reports were issued. Culture tubes with CPE were tested by the Dupont Herpchek enzyme immuno assay (EIA) to confirm the presence of herpes simplex virus (HSV). (3,4) 170 OB patient specimens were positive by culture and confirmed by EIA.There were also 63 cultures exhibiting CPE ressembling HSV which were negative by EIA testing, which failed to pass in fresh tissue culture cells or progress to more enhanced CPE in culture. These original specimens were screened by electron microscopy after direct ultracentrifugation employing the Beckman airfuge with the EM 90 rotor on to formvar carbon-coated 300 mesh copper grids and negatively stained with 2% PTA.
Structural integrity after cold storage of human epidermal model in hypothermosol: A correlative ultrastructural and fluorescent assay
Authors: Eichelberger Henry H. , Baust John G. , Van Buskirk Robert G. 
Published: June 2020

For research in cell differentiation and in vitro toxicology it is essential to provide a natural state of cell structure as a benchmark for interpreting results. Hypothermosol (Cryomedical Sciences, Rockville, MD) has proven useful in insuring the viability of synthetic human epidermis during cold-storage and in maintaining the epidermis’ ability to continue to differentiate following warming.Human epidermal equivalent, EpiDerm (MatTek Corporation, Ashland, MA) consisting of fully differentiated stratified human epidermal cells were grown on a microporous membrane. EpiDerm samples were fixed before and after cold-storage (4°C) for 5 days in Hypothermosol or skin culture media (MatTek Corporation) and allowed to recover for 7 days at 37°C. EpiDerm samples were fixed 1 hour in 2.5% glutaraldehyde in sodium cacodylate buffer (pH 7.2). A secondary fixation with 0.2% ruthenium tetroxide (Polysciences, Inc., Warrington, PA) in sodium cacodylate was carried out for 3 hours at 4°C. Other samples were similarly fixed, but with 1% Osmium tetroxide in place of ruthenium tetroxide. Samples were dehydrated through a graded acetone series, infiltrated with Spurrs resin (Polysciences Inc.) and polymerized at 70°C.
In vitro assembly of 2-D crystals from partially purified EA1 protein secreted by Bacillus anthracis strain Delta Sterne-1
Authors: Downs Mary Beth , Ribot Wilson , Farchaus Joseph W. 
Published: June 2020

Many bacteria possess surface layers (S-layers) that consist of a two-dimensional protein lattice external to the cell envelope. These S-layer arrays are usually composed of a single species of protein or glycoprotein and are not covalently linked to the underlying cell wall. When removed from the cell, S-layer proteins often reassemble into a lattice identical to that found on the cell, even without supporting cell wall fragments. S-layers exist at the interface between the cell and its environment and probably serve as molecular sieves that exclude destructive macromolecules while allowing passage of small nutrients and secreted proteins. Some S-layers are refractory to ingestion by macrophages and, generally, bacteria are more virulent when S-layers are present.When grown in rich medium under aerobic conditions, B. anthracis strain Delta Sterne-1 secretes large amounts of a proteinaceous extractable antigen 1 (EA1) into the growth medium. Immunocytochemistry with rabbit polyclonal anti-EAl antibody made against the secreted protein and gold-conjugated goat anti-rabbit IgG showed that EAI was localized at the cell surface (fig 1), which suggests its role as an S-layer protein.
Detection of atrial natriuretic peptides (ANP) in rat atria by immunogold-silver staining (IGSS)
Authors: Rufner Robyn , Hacker Gerhard W. , Forte Michele , Carson Nancyleigh E. , Xenachis Cristina , Gu Jiang 
Published: June 2020

The use of immunogold-silver staining (IGSS) to enhance label penetration and Localization for immunocytochemistry or in situ hybridization utilizing a variety of metallic salts has been documented. In this morphological study, the effects of silver acetate, silver lactate and silver nitrate were evaluated for immunogold-labeling of a trial natriuretic peptides (ANP) in rat right atria.Eight Wistar Kyoto retired breeders were sedated with pentobarbital, perfused with either 4% paraformaldehyde (LM) or Karnovsky's fixative (EM), and right atria were dissected, processed, embedded in paraffin or epon, respectively and sectioned according to conventional methods. For light microscopy, an indirect IGSS method according to Hacker (3) was performed. Paraffin sections on glass slides were washed in ddH2O, immersed in Lugol's iodine, washed in ddH2O and treated with 2.5% aqueous sodium thiosulfate for 20 sec. After additional washes in ddH2O and TBS-0.1% fish gelatin, 10% normal goat serum (PBS with 1% BSA) was applied for 20 min before an overnight incubation at 4°C with a polyclonal α-ANP primary antibody (Peninsula Labs, 1:1000 in TBS/BSA).
Three-Dimensional reconstruction of isolated centrosomes by automated electron tomography
Authors: Braunfeld M.B. , Moritz M. , Alberts B.M. , Sedat J.W. , Agard D.A. 
Published: June 2020

In animal cells, the centrosome functions as the primary microtubule organizing center (MTOC). As such the centrosome plays a vital role in determining a cell's shape, migration, and perhaps most importantly, its division. Despite the obvious importance of this organelle little is known about centrosomal regulation, duplication, or how it nucleates microtubules. Furthermore, no high resolution model for centrosomal structure exists.We have used automated electron tomography, and reconstruction techniques in an attempt to better understand the complex nature of the centrosome. Additionally we hope to identify nucleation sites for microtubule growth.Centrosomes were isolated from early Drosophila embryos. Briefly, after large organelles and debris from homogenized embryos were pelleted, the resulting supernatant was separated on a sucrose velocity gradient. Fractions were collected and assayed for centrosome-mediated microtubule -nucleating activity by incubating with fluorescently-labeled tubulin subunits. The resulting microtubule asters were then spun onto coverslips and viewed by fluorescence microscopy.
A rapid method for negative staining of fecal samples for diagnosis of viral-induced gastroenteritis by Transmission Electron Microscopy
Authors: Barrish J. P. , Hicks MJ , Leer LC , Hayes ES , Estes MK , Cubitt WD 
Published: June 2020

The most common etiologic agents for viral gastroenteritis are rotavirus, adenovirus, and members of the “small round structure virus” category (enterovirus, norwalk virus, calicivirus, astrovirus). A rapid diagnosis of viral infection can avoid unnecessary antibiotic therapy, extensive and costly medical workups and reduce the patient’s hospital stay. Pseudoreplication has been the preparation method of choice at many institutions, because of its concentrating effect on viral particles. Unfortunately, in addition to concentrating viral particles, this method also concentrates fecal debris, making the screening process quite difficult at times and time consuming.A rapid method of negative staining for viral particle identification in fecal samples is presented. The method first described by Cubitt et al has been modified, resulting in a reduction in the amount of debris and improved staining quality of the viral particles, while still concentrating the number of viral particles. This method requires less than 1 ml of stool specimen and may be prepared in less than three minutes.
Three-Dimensional reconstruction of protein bodies in developing maize endosperm using immunocytochemical techniques
Authors: Lending C. , Spinelli S. 
Published: June 2020

The storage proteins of maize (Zea mays L.), like many other cereals, are a group of alcohol-soluble proteins classified as prolamines. In maize these proteins comprise four structurally distinct types and are termed zeins. Zein synthesis is initiated in developing maize endosperm and continues until the seed reaches maturity (between approximately 12 to 50 days after pollination). Zeins are synthesized by polysomes bound to the rough endoplasmic reticulum (RER) and they accumulate as insoluble aggregates called protein bodies. Our previous studies have shown that the zeins are deposited within protein bodies in a defined order in normal genotypes. However, the actual three-dimensional organization of the zeins within protein bodies and the interactions that occur between the various zeins is not known. Our goal is to understand protein body organization and the interactions that occur between the various proteins.The four types of zeins are identified by their molecular weight after separation by SDS-PAGE, and are designated α-, β-, γ- and 6-zeins. The β-, γ-, and δ-zeins are all sulfur-rich proteins, while the α-zeins contain only low amounts of cysteine and methionine. These proteins can be individually detected by immunocytochemical staining of ultrathin sections. Additionally, ultrathin sections poststained with lead citrate and uranyl acetate demonstrate that there are two distinct regions that correlate with the immunocytochemical staining patterns.
An SEM and EDS study of secretions from the leaves of two halophytes
Authors: Naidoo Yougasphree , Gregory Michael A. , Naidoo Gonasagren 
Published: June 2020

Salt glands are secretory organs that occur in several angiosperm families including the Plumbaginaceae and Poaceae. They are particularly prevalent on the leaves of plants growing in saline habitats where by the secretion of ions, they prevent toxic levels of salts from accumulating in their tissues. Whether different species of halophytes containing morphologically disparate glands preferentially secrete different cocktails of ions is not known. This question was addressed by employing SEM and EDS to determine the morphology and elemental constituents of crystalline deposits over the salt glands of two species of salt marsh halophytes: Limonium scabrum (Thunb.) Kuntze (Plumbaginaceae) and Sporobolus virginicus (L.) Kunth (Poaceae).The two halophytes, collected from intertidal marshes and cultivated in a mixture of coarse sand and compost (3:1 by volume) were irrigated daily with tap water and fertilised every two weeks with full strength Hoagland’s nutrient solution. Plants were watered weekly for 3 months with 50 mol m−3 NaCl supplied as diluted sea water.
Scanning electron microscope study of brochosomes of cercopidae
Authors: Frost A. S. , Gardner J. S. , Nielson M. 
Published: June 2020

Brochosomes are minute granules secreted by specialized cells lining the lumen of Malphigian tubules in homopteran insects. They have been reported on two different homopteran families, the Cicadellidae (leaf-hoppers) and Membracidae (tree-hoppers), but they have never been reported on members of the family Cercopidae (froghoppers), although they have been suspected to be produced by other homopteran families. These three families all belong to the subgroup Auchenorryncha, and often coexist in the same habitat. Many theories have been proposed regarding brochosome function, but no conclusive results have ever been reported.Scanning electron microscopy was used to study legs of five species of Cercopidae. Brochosomes were observed on legs of Lepyronid batrachoidea (Fig. 1), Clastoptera brunnea (Fig. 2), Philaronia bilineata (Fig. 3), and Aphozphoza media (Fig. 4). None was found on Clastoptera juniperina. The shape of the brochosomes remained fairly consistent on all species except Clastoptera brunnea. Normally the structure was a polyhedron consisting of 12 pentagons and 20 hexagons, appearing hollow and nearly spherical. However, on Clastoptera brunnea, the brochosomes were larger and appeared to be more faceted.
Early ossicle growth in the regenerating disc of a brittle star
Authors: Shlepr M. , Turner R. L. 
Published: June 2020

Calcification in the echinoderms occurs within a limited-volume cavity enclosed by cytoplasmic extensions of the mineral depositing cells, the sclerocytes. The current model of this process maintains that the sheath formed from these cytoplasmic extensions is syncytial. Prior studies indicate that syncytium formation might be dependent on sclerocyte density and not required for calcification. This model further envisions that ossicles formed de novo nucleate and grow intracellularly until the ossicle effectively outgrows the vacuole. Continued ossicle growth occurs within the sheath but external to the cell membrane. The initial intracellular location has been confirmed only for elements of the echinoid tooth.The regenerating aboral disc integument of ophiophragmus filograneus was used to test the current echinoderm calcification model. This tissue is free of calcite fragments, thus avoiding questions of cellular engulfment, and ossicles are formed de novo. The tissue calcification pattern was followed by light microscopy in both living and fixed preparations.
Cytolysis caused by selenite uptake in the Bacillus subtilis
Authors: Yang W. H. , Yang J. R. 
Published: June 2020

The toxic effects of selenium compounds on animals are well documented. However, the cytotoxic effects of these compounds are not well known. Using electron microscopy and atomic absorption spectrophotometry, we studied the effect of sodium selenite on the fine structure of Bacillus subtilis.Bacteria were cultured with sodium selenite in LB broth at 38°C in a shaker culture system. At concentrations of selenite greater than 1mM, an inhibitory effect on bacterial growth was seen during the first few hours of culture. Using the atomic absorption assay, no uptake of selenite into bacteria during the lag phase was demonstrable. However, during the exponential growth phase, a marked increase in the selenium content of the cell pellet was measured along with a decrease in selenite in the supernatant (Figure 1). A reddish discoloration of the culture media was observed which corresponded with increasing selenite uptake by bacteria. After 24 hours in culture, an increase in particle density and viscosity of the culture media was measured which suggested fragmentation of the bacteria resulting in the release of DNA into the media.
Automated linescan analysis in the electron microprobe
Authors: Spinnler G. E. , Christenson D. , Nielsen C. H. 
Published: June 2020

Elemental linescan analysis in the electron microprobe (EMPA) or the SEM is useful for determining athe spatial distribution of elements. In many supported catalyst systems, the elemental distribution in pellets is important for their manufacture and performance. More important than determining the distribution of an individual pellet, is determining the distribution of many pellets in a sample to represent the lot. For this reason, multiple pellets must be analyzed. Various manufactures have systems that provide various degrees of automation for this type of analysis, however, there are no systems that allow complete unattended operation necessary to collect data on many samples.The hardware consists of a JEOL JXM 8600 with four wavelength spectrometers, an EDX detector and a NORAN Voyager. Special software was written to completely automate the linescan procedure from finding the catalyst pellet to printing the report. This software combines the imaging and other analytical functions of the Voyager with a search and identify routine that locates the catalyst pellets and determines the positions of the linescan in accordance with predetermined criteria.
Crystal tectonics: Novel routes to the ordered aggregation and self assembly of inorganic solids
Authors: Heywood Brigid R. 
Published: June 2020

Oriented materials attract considerable attention since thay have the potential to exhibit collective properties which exceed those of the isotropic species by several orders of magnitude. Much success has already been achieved with organic materials, e.g. liquid crystals, conducting polymers, but reliable protocols for the construction of organised crystal micro-architectures from inorganic solids have yet to be established. Given the potential advantages of translating molecular properties (optical, piezoelectric, catalytic) to the macroscopic scale strategies for the construction of hierarchical crystal assemblies, crystal tectonics, merit particular consideration.This crystal tectonics route to the synthesis of anisotropic inorganic materials remains entirely untested, but draws much of its inspiration from the study of deterministic self-organisation in biological systems. Such self-organisation relies on a series of highly specific “host-guest”, ligandreceptor type interactions (more typically cited examples of such include, enzyme-substrate-cofactor binding, antibody-antigen complexation, and triplet/base matching during polypeptide synthesis). The biogenic formation of hierarchical inorganic arrays, biomineralization, is remarkable not only for its control of crystallisation to yield solids of uniform size and unusual habit, but equally for the construction of elaborate functional micro-architectures from these biosolids.
Digital image acquisition, processing, and analysis in a polymer microscopy laboratory
Authors: Berry Vinod K. , Zhang Xiao 
Published: June 2020

In recent years it became apparent that we needed to improve productivity and efficiency in the Microscopy Laboratories in GE Plastics. It was realized that digital image acquisition, archiving, processing, analysis, and transmission over a network would be the best way to achieve this goal. Also, the capabilities of quantitative image analysis, image transmission etc. available with this approach would help us to increase our efficiency. Although the advantages of digital image acquisition, processing, archiving, etc. have been described and are being practiced in many SEM, laboratories, they have not been generally applied in microscopy laboratories (TEM, Optical, SEM and others) and impact on increased productivity has not been yet exploited as well.In order to attain our objective we have acquired a SEMICAPS imaging workstation for each of the GE Plastic sites in the United States. We have integrated the workstation with the microscopes and their peripherals as shown in Figure 1.
Using the atomic-force microscope to measure roughness characteristics of acetabular prosthetic shells used as hip-socket replacements
Authors: Williams Gary. , Jani Shilesh , Marchese-Ragona Silvio P. 
Published: June 2020

Total replacement of a diseased or traumatized hip joint is fast becoming one of the most common surgical procedures. The acetabular prosthetic shell is gaining widespread acceptance as a socket replacement for this procedure. Bio-materials research is providing a wealth of information regarding the life span, wear characteristics, and performance of these replacements, with a major clinical finding being the destruction of bone (osteolysis) at interfaces with the implants. Osteolysis is widely thought to be associated with the particulate wear debris released from articulation of the ball and socket. Formation and release of these particulates will depend not only on the relative motion occurring but also the surface roughness of the metal.In this study we used the Atomic Force Microscope (AFM) to compare the internal surface topography of two acetabular shells, one polished to a mirror finish and the other unpolished. The ultimate goal of the study was to determine whether a polished shell reduces the amount of wear and tear on the ultra-high molecular weight polyethylene (UHMWPE) insert that is fitted between the shell and the Femoral ball (Figure 1).
Energy-dispersive x-ray spectroscopy: Escape peaks of some biomaterials in the light-element region
Authors: Okerstrom Susan J. 
Published: June 2020

The phenomena of escape peaks is well known, but caution must be exercised to avoid confusion in peak identification. A summary of our experiences with certain escape peaks may be useful to others.In biomedical devices, metal components are often used in pure elemental form or as alloys with high concentrations of a particular element. Noble metals such as gold, platinum and iridium are commonly used in pacemaker components. These metals are body compatible, have high corrosion resistance and are good electrical conductors. Pacemaker electrodes are commonly made of platinum, used alone or alloyed with about 10% iridium. These materials are routinely analyzed using the scanning electron microscope (SEM) with energy-dispersive x-ray spectroscopy (EDS).While analyzing platinum electrodes at 20 keV with the Beryllium window open, it was noted that an unexpected peak was seen in the spectrum near where carbon would be expected at 0.310 keV. The same peak was noted when the Be window was closed (figure 1). It was first thought to be a detector problem.
Gas reaction chamber for gas-solid interaction studies by high-resolution TEM
Authors: Sharma Renu , Weiss Karl , McKelvy Michael , Glaunsinger William 
Published: June 2020

An environmental cell (E-cell) is a gas reaction chamber mounted inside an electron microscope column where thin solid samples can be observed under various gases (O2, H2, N2, NH3etc.) at selected temperatures. Even though the idea of having an E-cell incorporated in the microscope column is as old as transmission electron microscopy itself, recent developments in the instrumentation and designs of both the microscopes and E-cells have made it possible to obtain high resolution images (0.3-0.6 nm). We have used the differentially pumped model proposed by Swan to modify a PHILLIPS 400T transmission electron microscope for gas-solid studies.Figure la shows a side view cross section schematic of the E-cell fitted in the 9 mm gap between twin lens objective pole pieces. It consists of a small chamber with 200 and 400 μm apertures on sides a and a’ respectively. The walls are machined at the same angle as the pole pieces for an optimum fit to the conical exterior of the pole pieces and the cell is held firmly in place with o-rings (b).
Development of UHVTEM with UHV compatible goniometer and specimen-treatment chamber for surface studies
Authors: Kondo Y. , Kobayashi H. , Ishibashi Y. , Kasai T. , Nunome H. , Honda H. , Kersker M. , Ishida Y. 
Published: June 2020

JEOL has previously been involved in the development of various ultrahigh vacuum transmission electron microscopes (UHVTEM). Interest in UHVTEM is increasing, and for higher vacuum, higher resolution and specimen treatment capability, we have developed a new type of UHVTEM, the JEM- 2000FXVII. This paper reports the construction and performance of this new UHVTEM including a newly developed UHV compatible goniometer, specimen treatment chamber and various types of specimen heating holders.Figure 1 shows the overview of the JEM-2000FXVII with a specimen pre-treatment chamber (PTC) and UHV compatible goniometer. The PTC consists of two chambers: one for specimen treatment and the other for pre-evacuation. Specimen treatment is accomplished by using any of 7 attachment ports (six 70ϕ ICFs and a 152ϕ ICFs) in the PTC (Fig. 2). The PTC is attached directly to the TEM column and quick exchange of the specimen from PTC to column is thus possible. In-situ observation is also possible using the three attachment ports (70ϕ ICFs) in the column.