Self-assembly of chlorophenols in water

PubMed Central

Rogalska, Ewa; Rogalski, Marek; Gulik-Krzywicki, Tadeusz; Gulik, Annette; Chipot, Christophe

1999-01-01

In saturated solutions of some di- and trichlorophenols, structures with complex morphologies, consisting of thin, transparent sheets often coiling into helices and ultimately twisting into filaments, were observed under the optical microscope. Freeze-fracture electron microscopy, x-ray diffraction, phase diagrams, and molecular modeling were performed to elucidate the observed phenomena. Here, we present evidence that the chlorophenols studied, when interacting with water, self-assemble into bilayers. The fact that some chlorophenols form the same supramolecular structures as those described previously for structurally nonrelated surfactants sheds light on the mechanisms of self-assembly. PMID:10359753

Drying techniques for the visualisation of agarose-based chromatography media by scanning electron microscopy.

PubMed

Nweke, Mauryn C; Turmaine, Mark; McCartney, R Graham; Bracewell, Daniel G

2017-03-01

The drying of chromatography resins prior to scanning electron microscopy is critical to image resolution and hence understanding of the bead structure at sub-micron level. Achieving suitable drying conditions is especially important with agarose-based chromatography resins, as over-drying may cause artefact formation, bead damage and alterations to ultrastructural properties; and under-drying does not provide sufficient resolution for visualization under SEM. This paper compares and contrasts the effects of two drying techniques, critical point drying and freeze drying, on the morphology of two agarose based resins (MabSelect™/d w ≈85 µm and Capto™ Adhere/d w ≈75 µm) and provides a complete method for both. The results show that critical point drying provides better drying and subsequently clearer ultrastructural visualization of both resins under SEM. Under this protocol both the polymer fibers (thickness ≈20 nm) and the pore sizes (diameter ≈100 nm) are clearly visible. Freeze drying is shown to cause bead damage to both resins, but to different extents. MabSelect resin encounters extensive bead fragmentation, whilst Capto Adhere resin undergoes partial bead disintegration, corresponding with the greater extent of agarose crosslinking and strength of this resin. While freeze drying appears to be the less favorable option for ultrastructural visualization of chromatography resin, it should be noted that the extent of fracturing caused by the freeze drying process may provide some insight into the mechanical properties of agarose-based chromatography media. © 2017 The Authors. Biotechnology Journal published by WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Environmental Scanning Electron Microscope Imaging of Vesicle Systems.

PubMed

Perrie, Yvonne; Ali, Habib; Kirby, Daniel J; Mohammed, Afzal U R; McNeil, Sarah E; Vangala, Anil

2017-01-01

The structural characteristics of liposomes have been widely investigated and there is certainly a strong understanding of their morphological characteristics. Imaging of these systems, using techniques such as freeze-fracturing methods, transmission electron microscopy, and cryo-electron imaging, has allowed us to appreciate their bilayer structures and factors which can influence this. However, there are few methods which all us to study these systems in their natural hydrated state; commonly the liposomes are visualized after drying, staining, and/or fixation of the vesicles. Environmental Scanning Electron Microscopy (ESEM) offers the ability to image a liposome in its hydrated state without the need for prior sample preparation. Within our studies we were the first to use ESEM to study liposomes and niosomes and we have been able to dynamically follow the hydration of lipid films and changes in liposome suspensions as water condenses on to, or evaporates from, the sample in real time. This provides insight into the resistance of liposomes to coalescence during dehydration, thereby providing an alternative assay of liposome formulation and stability.

Cell-cell interactions of isolated and cultured oligodendrocytes: formation of linear occluding junctions and expression of peculiar intramembrane particles.

PubMed

Massa, P T; Szuchet, S; Mugnaini, E

1984-12-01

Oligodendrocytes were isolated from lamb brain. Freshly isolated cells and cultured cells, either 1- to 4-day-old unattached or 1- to 5-week-old attached, were examined by thin section and freeze-fracture electron microscopy. Freeze-fracture of freshly isolated oligodendrocytes showed globular and elongated intramembrane particles similar to those previously described in oligodendrocytes in situ. Enrichment of these particles was seen at sites of inter-oligodendrocyte contact. Numerous gap junctions and scattered linear tight junctional arrays were apparent. Gap junctions were connected to blebs of astrocytic plasma membrane sheared off during isolation, whereas tight junctions were facing extracellular space or blebs of oligodendrocytic plasma membrane. Thin sections of cultured, unattached oligodendrocytes showed rounded cell bodies touching one another at points without forming specialized cell junctions. Cells plated on polylysine-coated aclar dishes attached, emanated numerous, pleomorphic processes, and expressed galactocerebroside and myelin basic protein, characteristic markers for oligodendrocytes. Thin sections showed typical oligodendrocyte ultrastructure but also intermediate filaments not present in unattached cultures. Freeze-fracture showed intramembrane particles similar to but more numerous, and with a different fracture face repartition, than those seen in oligodendrocytes, freshly isolated or in situ. Gap junctions were small and rare. Apposed oligodendrocyte plasma membrane formed linear tight junctions which became more numerous with time in culture. Thus, cultured oligodendrocytes isolated from ovine brains develop and maintain features characteristic of mature oligodendrocytes in situ and can be used to explore formation and maintenance of tight junctions and possibly other classes of cell-cell interactions important in the process of myelination.

Visualization of pores (export sites) correlated with cellulose production in the envelope of the gram-negative bacterium Acetobacter xylinum.

PubMed

Zaar, K

1979-03-01

The Gram-negative bacterium Acetobacter xylinum assembles a cellulse ribbon composed of a number of microfibrils in the longitudinal axis of its envelope. The zone of ribbon assembly was investigated by freeze-etch electron microscopy. Freeze-etching revealed, beneath the cellulose ribbons, a linear array of pores on the lipopolysaccharide membrane. These pores have a rim diameter of 120--150 A and a central hole or deepening of approximately 35 A. The axes of pore arrays closely coincide with linear arrays of 100 A particles on the E- and P-faces of the fractured lipopolysaccharide membranes. Pores and particles in the lipopolysaccharide membrane are probably congruent. The pores are hypothesized to be the export sites (penetration sites) for cellulose.

Characterization of new DOPC/DHPC platform for dermal applications.

PubMed

Rodríguez, Gelen; Rubio, Laia; Barba, Clara; López-Iglesias, Carmen; de la Maza, Alfons; López, Olga; Cócera, Mercedes

2013-05-01

Systems formed by mixtures of the phospholipids dioleoylphosphatidylcholine (DOPC) and dihexanoylphosphatidylcholine (DHPC) were characterized by use of differential scanning calorimetry, small angle X-ray scattering and two electron-microscopy techniques, freeze fracture electron microscopy and cryogenic transmission electron microscopy. These techniques allowed for the determination of the size, morphology, structural topology, self-assembly and thermotropic behavior of the nanostructures present in the mixtures. The interaction between the two phospholipids provides curvatures, irregularities and the increase of thickness and flexibility in the membrane. These effects led to the formation of different aggregates with a differential distribution of both phospholipids. The effect of these systems on the skin in vivo was evaluated by measurement of the biophysical skin parameters. Our results show that the DOPC/DHPC application induces a decrease in the permeability and in the hydration of the tissue. These effects in vivo are related to different microstructural changes promoted by these systems in the skin in vitro, published in a recent work. The fundamental biophysical analyses of DOPC/DHPC systems contribute to our understanding of the mechanisms that govern their interaction with the skin.

High-pressure freezing and freeze substitution of Arabidopsis for electron microscopy.

PubMed

Austin, Jotham R

2014-01-01

The objectives of electron microscopy ultrastructural studies are to examine cellular architecture and relate the cell's structural machinery to dynamic functional roles. This aspiration is difficult to achieve if specimens have not been adequately preserved in a "living state"; hence specimen preparation is of the utmost importance for the success of any electron micrographic study. High-pressure freezing (HPF)/freeze substitution (FS) has long been recognized as the primer technique for the preservation of ultrastructure in biological samples. In most cases a basic HPF/freeze substitution protocol is sufficient to obtain superior ultrastructural preservation and structural contrast, which allows one to use more advanced microscopy techniques such as 3D electron tomography. However, for plant tissues, which have a thick cell wall, large water-filled vacuoles, and air spaces (all of which are detrimental to cryopreservation), these basic HPF/FS protocols often yield undesirable results. In particular, ice crystal artifacts and the staining of membrane systems are often poorly or negatively stained, which make 3D segmentation of a tomogram difficult. To overcome these problems, various aspects of the HPF/FS protocol can be altered, including the cryo-filler(s) used, freeze substitution cocktail, and the resin infiltration process. This chapter will describe these modifications for the preparation of plant tissues for routine electron microscopic studies, immunocytochemistry, and 3D tomographic electron imaging.

A SIMPLE FREEZE-FRACTURE REPLICATION METHOD FOR ELECTRON MICROSCOPY

PubMed Central

Bullivant, Stanley; Ames, Adelbert

1966-01-01

A simple method to achieve results similar to the freeze-etching technique of Moor et al. (1961) is described. The frozen tissue is cut under liquid nitrogen with a razor blade outside the evaporator rather than inside with a cooled microtome. The conditions of the experiment do not favor sublimation, and it is proposed that the structure of the replica be explained by local faults in the cleavage plane which leaves structures, such as membranes, standing above the ice. Micrographs of replicas of glycerol-protected frozen small intestine of mouse prepared by the method are presented and the structural details they show are discussed. The problem of vapor-deposited contamination is discussed. It is concluded that this is a practical method for obtaining electron micrographs that are relatively free of artifact, and that further improvements may be expected from the use of rapidly frozen fresh tissue and a clean vacuum system, possibly of the ion-pumped type. PMID:5962938

Freeze-fracture scanning electron microscopy of Lemna minor L. (duckweed)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Echlin, P.; Pawley, J.B.; Hayes, T.L.

1979-01-01

A detailed study has been carried out on the frozen fracture faces of plant material. Roots of Lemna minor are encapsulated in different nonpenetrating polymeric cryoprotectants, rapidly cooled in melting nitrogen and transferred to the pre-cooled cold stage of the AMR Biochamber. The technique has been used to follow the course of development of the phloem tissue in the root tip. These studies have shown that the phloem parenchyma appears to develop in a regular sequence. Unetched surfaces are virtually featureless, and it is necessary to remove a surface layer of water in order to visualize the biological structure. Themore » amount of water sublimed from the fractured surface is a function of both the time of etching and the water binding capacity of the cell contents. It is not possible to etch cells infiltrated with a penetrating cryoprotectant as the glycerol-water eutectic is stable at low temperatures and no water is lost from the fractured surface. Several distinct stages have been observed during the etching process.« less

DOE Office of Scientific and Technical Information (OSTI.GOV)

Phuthong, Witchukorn; Huang, Zubin; Wittkopp, Tyler M.

To investigate the dynamics of photosynthetic pigment-protein complexes in vascular plants at high resolution in an aqueous environment, membrane-protruding oxygen-evolving complexes (OECs) associated with photosystem II (PSII) on spinach ( Spinacia oleracea) grana membranes were examined using contact mode atomic force microscopy. This study represents, to our knowledge, the first use of atomic force microscopy to distinguish the putative large extrinsic loop of Photosystem II CP47 reaction center protein (CP47) from the putative oxygen-evolving enhancer proteins 1, 2, and 3 (PsbO, PsbP, and PsbQ) and large extrinsic loop of Photosystem II CP43 reaction center protein (CP43) in the PSII-OEC extrinsicmore » domains of grana membranes under conditions resulting in the disordered arrangement of PSII-OEC particles. Moreover, we observed uncharacterized membrane particles that, based on their physical characteristics and electrophoretic analysis of the polypeptides associated with the grana samples, are hypothesized to be a domain of photosystem I that protrudes from the stromal face of single thylakoid bilayers. Furthermore, our results are interpreted in the context of the results of others that were obtained using cryo-electron microscopy (and single particle analysis), negative staining and freeze-fracture electron microscopy, as well as previous atomic force microscopy studies.« less

The Use of Contact Mode Atomic Force Microscopy in Aqueous Medium for Structural Analysis of Spinach Photosynthetic Complexes

DOE PAGES

Phuthong, Witchukorn; Huang, Zubin; Wittkopp, Tyler M.; ...

2015-07-28

To investigate the dynamics of photosynthetic pigment-protein complexes in vascular plants at high resolution in an aqueous environment, membrane-protruding oxygen-evolving complexes (OECs) associated with photosystem II (PSII) on spinach ( Spinacia oleracea) grana membranes were examined using contact mode atomic force microscopy. This study represents, to our knowledge, the first use of atomic force microscopy to distinguish the putative large extrinsic loop of Photosystem II CP47 reaction center protein (CP47) from the putative oxygen-evolving enhancer proteins 1, 2, and 3 (PsbO, PsbP, and PsbQ) and large extrinsic loop of Photosystem II CP43 reaction center protein (CP43) in the PSII-OEC extrinsicmore » domains of grana membranes under conditions resulting in the disordered arrangement of PSII-OEC particles. Moreover, we observed uncharacterized membrane particles that, based on their physical characteristics and electrophoretic analysis of the polypeptides associated with the grana samples, are hypothesized to be a domain of photosystem I that protrudes from the stromal face of single thylakoid bilayers. Furthermore, our results are interpreted in the context of the results of others that were obtained using cryo-electron microscopy (and single particle analysis), negative staining and freeze-fracture electron microscopy, as well as previous atomic force microscopy studies.« less

Fabrication and characterisation of a novel biomimetic anisotropic ceramic/polymer-infiltrated composite material.

PubMed

Al-Jawoosh, Sara; Ireland, Anthony; Su, Bo

2018-04-10

To fabricate and characterise a novel biomimetic composite material consisting of aligned porous ceramic preforms infiltrated with polymer. Freeze-casting was used to fabricate and control the microstructure and porosity of ceramic preforms, which were subsequently infiltrated with 40-50% by volume UDMA-TEGDMA polymer. The composite materials were then subjected to characterisation, namely density, compression, three-point bend, hardness and fracture toughness testing. Samples were also subjected to scanning electron microscopy and computerised tomography (Micro-CT). Three-dimensional aligned honeycomb-like ceramic structures were produced and full interpenetration of the polymer phase was observed using micro-CT. Depending on the volume fraction of the ceramic preform, the density of the final composite ranged from 2.92 to 3.36g/cm 3 , compressive strength ranged from 206.26 to 253.97MPa, flexural strength from 97.73 to 145.65MPa, hardness ranged from 1.46 to 1.62GPa, and fracture toughness from 3.91 to 4.86MPam 1/2 . Freeze-casting provides a novel method to engineer composite materials with a unique aligned honeycomb-like interpenetrating structure, consisting of two continuous phases, inorganic and organic. There was a correlation between the ceramic fraction and the subsequent, density, strength, hardness and fracture toughness of the composite material. Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.

Investigation on the modification behavior of A356.2 alloy with Yb-La composite modifier

NASA Astrophysics Data System (ADS)

Zhang, Shaochen; Leng, Jinfeng; Wang, Zhibin; Wang, Qi; Shao, Yuewen; Teng, Xinying

2018-01-01

In this work, an investigation was conducted to evaluate the influence of the Yb + La addition on the microstructures and mechanical properties of A356.2 alloy. The results show that the the grain size was refined and morphological structure of Si changed from needle-like to fine spheroidal particle by adding Yb + La. The eutectic temperature decreased from 589 °C to 581 °C and the freezing range of primary α-Al become wider after adding Yb + La. The mechanical properties showed great improvement after T6 heat treatment. The ultimate tensile strength and microhardness of the composites were increased to 279.0 MPa and 99.2 HB after T6 heat treatment which increased by 28.2% and 47.3%, respectively. The fracture mechanism was analyzed using the scanning electron microscopy and proved to be a transgranular/intergranular fracture modes.

Freeze-fracture studies of photoreceptor membranes: new observations bearing upon the distribution of cholesterol

PubMed Central

1983-01-01

We performed electron microscopy of replicas from freeze-fractured retinas exposed during or after fixation to the cholesterol-binding antibiotic, filipin. We observed characteristic filipin-induced perturbations throughout the disk and plasma membranes of retinal rod outer segments of various species. It is evident that a prolonged exposure to filipin in fixative enhances rather than reduces presumptive cholesterol detection in the vertebrate photoreceptor cell. In agreement with the pattern seen in our previous study (Andrews, L.D., and A. I. Cohen, 1979, J. Cell Biol., 81:215-228), filipin- binding in membranes exhibiting particle-free patches seemed largely confined to these patches. Favorably fractured photoreceptors exhibited marked filipin-binding in apical inner segment plasma membrane topologically confluent with and proximate to the outer segment plasma membrane, which was comparatively free of filipin binding. A possible boundary between these differing membrane domains was suggested in a number of replicas exhibiting lower filipin binding to the apical plasma membrane of the inner segment in the area surrounding the cilium. This area contains a structure (Andrews, L. D., 1982, Freeze- fracture studies of vertebrate photoreceptors, In Structure of the Eye, J. G. Hollyfield and E. Acosta Vidrio, editors, Elsevier/North-Holland, New York, 11-23) that resembles the active zones of the nerve terminals for the frog neuromuscular junction. These observations lead us to hypothesize that these structures may function to direct vesicle fusion to occur near them, in a domain of membrane more closely resembling outer than inner segment plasma membrane. The above evidence supports the views that (a) all disk membranes contain cholesterol, but the particle-free patches present in some disks trap cholesterol from contiguous particulate membrane regions; (b) contiguous inner and outer segment membranes may greatly differ in cholesterol content; and (c) the suggested higher cholesterol in the inner segment than in the outer segment plasma membrane may help direct newly inserted photopigment molecules to the outer segment. PMID:6411740

Scanning electron microscope fractography in failure analysis of steels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wouters, R.; Froyen, L.

1996-04-01

For many failure cases, macroscopic examination of the fracture surface permits discrimination of fatigue fractures from overload fractures. For clarifying fatigue fractures, the practical significance of microfractography is limited to an investigation of the crack initiation areas. Scanning electron microscopy is successfully used in tracing local material abnormalities that act as fatigue crack initiators. The task for the scanning electron microscope, however, is much more substantial in failure analysis of overload fractures, especially for steels. By revealing specific fractographic characteristics, complemented by information about the material and the loading conditions, scanning electron microscopy provides a strong indication of the probablemore » cause of failure. A complete dimple fracture is indicative of acceptable bulk material properties; overloading, by subdimensioning or excessive external loading, has to be verified. The presence of cleavage fracture makes the material properties questionable if external conditions causing embrittlement are absent. Intergranular brittle fracture requires verification of grain-boundary weakening conditions--a sensitized structure, whether or not combined with a local stress state or a specific environment. The role of scanning electron microscopy in failure analysis is illustrated by case histories of the aforementioned fracture types.« less

An ultrastructural analysis of the epithelial-fiber interface (EFI) in primate lenses.

PubMed

Kuszak, J R; Novak, L A; Brown, H G

1995-11-01

The purpose of this study was to conduct a comprehensive ultrastructural analysis of the epithelial-fiber interface (EFI) in normal adult primate (Macaque nemestrina and fascicularis; 6-9 years old, n = 10) lenses. Scanning electron microscopy (SEM) was used to initially characterize the gross size, shape and three-dimensional organization of central zone (cz) epithelial cells and the anterior ends of elongating fibers beneath these cells. This fiducial information was essential to properly orient lens pieces in freeze fracture specimen carriers for the production of replicas with unambiguously identifiable EFI. Transmission electron microscopy (TEM) of replicas and thin-sectioned material were used to ultrastructurally analyse the cz EFI. TEM thin-sectioned material was also used to ultrastructurally analyse the pregerminative (pgz), germinative (gz) and transitional zone (tz) EFI. Correlative SEM and TEM of cz EFI components revealed that the apical membrane of both epithelial and elongating fiber cells were irregularly polygonal in shape, and aligned in parallel as smooth, concave-convex surfaces. However, whereas epithelial cell apical surfaces had minimal size variation, elongating fibers were larger and considerably variable in size. Quantitative analysis of > 10000 micron2 cz elongating fiber apical surfaces failed to detect any gap junctions defined in freeze fracture replicas as complementary aggregates of transmembrane proteins (connexons) conjoined across a narrowed extracellular space. However, a comparable frequency of vesicular events was noted in this region as quantified previously in adult and embryonic chick lens. Correlative TEM analysis > 1500 linear micrometers of thin-sectioned EFI from this region confirmed the presence of epithelial-epithelial gap junctions, elongating fiber-elongating fiber gap junctions, and an extreme paucity of epithelial-elongating fiber gap junctions. In contrast, TEM analysis of > 1000 linear micrometers of thin-sectioned pgz, gz and tz EFI, confirmed the presence of epithelial-epithelial gap junctions, elongating fiber-elongating fiber gap junctions, numerous epithelial-elongating fiber adherens junctions and a few epithelial-elongating fiber gap junctions. Thus, the results of this and previous quantitative morphological and physiological studies (electronic and dye coupling) demonstrate that there is limited coupling between cz epithelial cells and underlying elongating fibers. Furthermore, the absence of gap junctional plaques in cz EFI freeze-fracture replicas and either pentalaminar or septalaminar profiles in correlative thin-sections, suggests that this limited coupling could be mediated via isolated gap junction channels. However, the results of this and previous quantitative studies further show that a greater degree of coupling exists across the pgz, gz and tz regions of the EFI and that this coupling is likely to be mediated by gap junction plaques. Finally, this and other studies continue to demonstrate that transcytotic processes play a role in lens physiology at the EFI.

The application of scanning electron microscopy to fractography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brooks, C.R.; McGill, B.L.

1994-10-01

Many failures involve fracture, and determination of the fracture process is a key factor in understanding the failure. This is frequently accomplished by characterizing the topography of the fracture surface. Scanning electron microscopy has a prominent role in fractography due to three features of the scanning electron microscope (SEM): high resolution, great depth of field, and the ability to obtain chemical information via analysis of the X-rays generated by the electrons. A qualitative treatment is presented of the interaction of electrons with a sample and the effect of the SEM operating parameters on image formation, quality, and X-ray analysis. Fractographsmore » are presented to illustrate these features of scanning electron microscopy and to illustrate the limitations and precautions in obtaining fractographs and x-ray analyses. The review is concluded with examples of fracture surface features of metallic, ceramic, and polymeric materials.« less

Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy

PubMed Central

HÖHN, K.; FUCHS, J.; FRÖBER, A.; KIRMSE, R.; GLASS, B.; ANDERS‐ÖSSWEIN, M.; WALTHER, P.; KRÄUSSLICH, H.‐G.

2015-01-01

Summary In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV‐pulsed mature human dendritic cells. PMID:25786567

Cellulose biosynthesis in Acetobacter xylinum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lin, F.C.

1988-01-01

Time-lapse video microscopy has shown periodic reversals during the synthesis of cellulose. In the presence of Congo Red, Acetobacter produces a band of fine fibrils. The direction of cell movement is perpendicular to the longitudinal axis of cell, and the rate of movement was decreased. A linear row of particles, presumably the cellulose synthesizing complexes, was found on the outer membrane by freeze-fracture technique. During the cell cycle, the increase of particles in linear row, the differentiation to four linear rows and the separation of the linear rows have been observed. A digitonin-solubilized cellulose synthase was prepared from A. xylinum,more » and incubated under conditions known to lead to active in vitro synthesis of 1,4-{beta}-D-glucan polymer. Electron microscopy revealed that clusters of fibrils were assembled within minutes. Individual fibrils are 17 {plus minus} 2 angstroms in diameter. Evidence for the cellulosic composition of newly synthesized fibrils was based on incorporation of tritium from UDP-({sup 3}H) glucose binding of gold-labeled cellobiohydrolase, and an electron diffraction pattern identified as cellulose II polymorph instead of cellulose I.« less

Preservation of protein fluorescence in embedded human dendritic cells for targeted 3D light and electron microscopy.

PubMed

Höhn, K; Fuchs, J; Fröber, A; Kirmse, R; Glass, B; Anders-Össwein, M; Walther, P; Kräusslich, H-G; Dietrich, C

2015-08-01

In this study, we present a correlative microscopy workflow to combine detailed 3D fluorescence light microscopy data with ultrastructural information gained by 3D focused ion beam assisted scanning electron microscopy. The workflow is based on an optimized high pressure freezing/freeze substitution protocol that preserves good ultrastructural detail along with retaining the fluorescence signal in the resin embedded specimens. Consequently, cellular structures of interest can readily be identified and imaged by state of the art 3D confocal fluorescence microscopy and are precisely referenced with respect to an imprinted coordinate system on the surface of the resin block. This allows precise guidance of the focused ion beam assisted scanning electron microscopy and limits the volume to be imaged to the structure of interest. This, in turn, minimizes the total acquisition time necessary to conduct the time consuming ultrastructural scanning electron microscope imaging while eliminating the risk to miss parts of the target structure. We illustrate the value of this workflow for targeting virus compartments, which are formed in HIV-pulsed mature human dendritic cells. © 2015 The Authors Journal of Microscopy © 2015 Royal Microscopical Society.

Controlled microaspiration for high-pressure freezing: a new method for ultrastructural preservation of fragile and sparse tissues for TEM and electron tomography

PubMed Central

Triffo, W. J.; Palsdottir, H.; McDonald, K. L.; Lee, J. K.; Inman, J. L.; Bissell, M. J.; Raphael, R. M.; Auer, M.

2009-01-01

Summary High-pressure freezing is the preferred method to prepare thick biological specimens for ultrastructural studies. However, the advantages obtained by this method often prove unattainable for samples that are difficult to handle during the freezing and substitution protocols. Delicate and sparse samples are difficult to manipulate and maintain intact throughout the sequence of freezing, infiltration, embedding and final orientation for sectioning and subsequent transmission electron microscopy. An established approach to surmount these difficulties is the use of cellulose microdialysis tubing to transport the sample. With an inner diameter of 200 µm, the tubing protects small and fragile samples within the thickness constraints of high-pressure freezing, and the tube ends can be sealed to avoid loss of sample. Importantly, the transparency of the tubing allows optical study of the specimen at different steps in the process. Here, we describe the use of a micromanipulator and microinjection apparatus to handle and position delicate specimens within the tubing. We report two biologically significant examples that benefit from this approach, 3D cultures of mammary epithelial cells and cochlear outer hair cells. We illustrate the potential for correlative light and electron microscopy as well as electron tomography. PMID:18445158

Geometry of phase-separated domains in phospholipid bilayers by diffraction-contrast electron microscopy.

PubMed Central

Hui, S W

1981-01-01

The sizes and shapes of solidus (gel) phase domains in the hydrated molecular bilayers of dilauroylphosphatidylcholine/dipalmitoylphasphatidylcholine (DLPC/DPPC) (1:1) and phosphatidylserine (PS)/DPPC (1:2) are visualized directly by low dose diffraction-contrast electron microscopy. The temperature and humidity of the bilayers are controlled by an environmental chamber set in an electron microscope. The contrast between crystalline domains is enhanced by electron optical filtering of the diffraction patterns of the bilayers. The domains are seen as a patchwork in the plane of the bilayer, with an average width of 0.2-0.5 micrometer. The percentage of solidus area measured from diffraction-contrast micrographs at various temperatures agrees in general with those depicted by known phase diagrams. The shape and size of the domains resemble those seen by freeze-fracture in multilamellar vesicles. Temperature-related changes in domain size and in phase boundary per unit area are more pronounced in the less miscible DLPC/DPPC mixture. No significant change in these geometric parameters with temperature is found in the PS/DPPC mixture. Mapping domains by their molecular diffraction signals not only verifies the existance of areas of different molecular packing during phase separation but also provides a quantitative measurement of structural boundaries and defects in lipid bilayers. Images FIGURE 1 FIGURE 3 FIGURE 6 PMID:6894707

Chiral lyotropic chromonic liquid crystals composed of disodium cromoglycate doped with water-soluble chiral additives.

PubMed

Shirai, Tatsuya; Shuai, Min; Nakamura, Keita; Yamaguchi, Akihiro; Naka, Yumiko; Sasaki, Takeo; Clark, Noel A; Le, Khoa V

2018-02-28

We investigated the pitches of cholesteric liquid crystals prepared by mixing disodium cromoglycate (DSCG) in water with 5 different water-soluble chiral additives. The measurements are based on the Grandjean-Cano wedge cell method. Overall, the twisting effect is weak, and the shortest pitch of 2.9 ± 0.2 μm is obtained using trans-4-hydroxy-l-proline, by which the cholesteric sample is iridescent at certain viewing angles. Freeze-fracture transmission electron microscopy (FFTEM) was also performed for the first time on both the nematic and cholesteric phases, revealing that stacked chromonic aggregates are very long, up to a few hundred nm, which explains why cholesteric chromonic liquid crystals hardly have pitches in the visible wavelength region.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kononchik, Joseph P.; Vancini, Ricardo; Brown, Dennis T., E-mail: dennis_brown@ncsu.edu

Sindbis Virus (SV), the prototype alphavirus in the family togaviridae, infects both mammalian and insect cells. The ability of SV to infect cells possessing significantly different biochemical environments suggests that there may be a common mode of entry into each cell type. Previous studies show that up to 4 h post infection cells are permeable to small ions and alpha sarcin suggesting that the plasma membrane is compromised as infection takes place. Thin-section electron microscopy has also shown SV to bind to the plasma membrane and lose its electron dense core through a pore like structure developed upon interaction ofmore » the virus with the cell surface. Using freeze-fracture replicas, thin-sections and antibody labeling the data presented herein show virus associated with intramembrane particles on mosquito cells. These data suggest that the intramembrane particles associated with SV may be part of the pore structure consisting of virus proteins and cell receptor.« less

The electron microscope appearance of the subchondral bone plate in the human femoral head in osteoarthritis and osteoporosis

PubMed Central

LI, BAOHUA; MARSHALL, DEBORAH; ROE, MARTIN; ASPDEN, RICHARD M.

1999-01-01

The subchondral bone plate supports the articular cartilage in diarthrodial joints. It has a significant mechanical function in transmitting loads from the cartilage into the underlying cancellous bone and has been implicated in the destruction of cartilage in osteoarthritis (OA) and its sparing in osteoporosis (OP), but little is known of its composition, structure or material properties. This study investigated the microscopic appearance and mineral composition of the subchondral bone plate in femoral heads from patients with OA or OP to determine how these correspond to changes in composition and stiffness found in other studies. Freeze-fractured full-depth samples of the subchondral bone plate from the femoral heads of patients with osteoarthritis, osteoporosis or a matched control group were examined using back scattered and secondary emission scanning electron microscopy. Other samples were embedded and polished and examined using back-scattered electron microscopy and electron probe microanalysis. The appearances of the samples from the normal and osteoporotic patients were very similar, with the subchondral bone plate overlayed by a layer of calcified cartilage. Osteoporotic samples presented a more uniform fracture surface and the relative thicknesses of the layers appeared to be different. In contrast, the OA bone plate appeared to be porous and have a much more textured surface. There were occasional sites of microtrabecular bone formation between the trabeculae of the underlying cancellous bone, which were not seen in the other groups, and more numerous osteoclast resorption pits. The calcified cartilage layer was almost absent and the bone plate was apparently thickened. The appearance of the osteoarthritic subchondral bone plate was, therefore, considerably different from both the normal and the osteoporotic, strongly indicative of abnormal cellular activity. PMID:10473297

Freeze drying vs microwave drying-methods for synthesis of sinteractive thoria powders

NASA Astrophysics Data System (ADS)

Annie, D.; Chandramouli, V.; Anthonysamy, S.; Ghosh, Chanchal; Divakar, R.

2017-02-01

Thoria powders were synthesized by oxalate precipitation from an aqueous solution of the nitrate. The filtered precipitates were freeze dried or microwave dried before being calcined at 1073 K. The thoria powders obtained were characterized for crystallite size, specific surface area, bulk density, particle size distribution and residual carbon. Microstructure of the product was studied using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Sinterability of the synthesized powders was studied by measuring the density of the sintered compacts. Powders that can be consolidated and sintered to densities ∼96% theoretical density (TD) at 1773 K were obtained.

Histophilus somni biofilm formation in cardiopulmonary tissue of the bovine host following respiratory challenge.

PubMed

Sandal, Indra; Shao, Jian Q; Annadata, Satish; Apicella, Michael A; Boye, Mette; Jensen, Tim K; Saunders, Geoffrey K; Inzana, Thomas J

2009-02-01

Biofilms form in a variety of host sites following infection with many bacterial species. However, the study of biofilms in a host is hindered due to the lack of protocols for the proper experimental investigation of biofilms in vivo. Histophilus somni is an agent of respiratory and systemic diseases in bovines, and readily forms biofilms in vitro. In the present study the capability of H. somni to form biofilms in cardiopulmonary tissue following experimental respiratory infection in the bovine host was examined by light microscopy, transmission electron microscopy, immunoelectron microscopy of ultrathin cryosections, scanning electron microscopy of freeze-fractured samples, and fluorescent in situ hybridization. Biofilms were evident and most prominent in the myocardium, and were associated with a large amount of amorphous extracellular material. Furthermore, Pasteurella multocida was often cultured with H. somni from heart and lung samples. Transposon mutagenesis of H. somni strain 2336 resulted in the generation of mutants that expressed more or less biofilm than the parent strain. Six mutants deficient in biofilm formation had an insertion in the gene encoding for a homolog of filamentous haemagglutinin (FHA), predicted to be involved in attachment. Thus, this investigation demonstrated that H. somni is capable of forming a biofilm in its natural host, that such a biofilm may be capable of harboring other bovine respiratory disease pathogens, and that the genes responsible for biofilm formation can be identified by transposon mutagenesis.

Candidacidal effects of two antimicrobial peptides: histatin 5 causes small membrane defects, but LL-37 causes massive disruption of the cell membrane

PubMed Central

2005-01-01

The effects of antimicrobial peptides on artificial membranes have been well-documented; however, reports on the ultrastructural effects on the membranes of micro-organisms are relatively scarce. We compared the effects of histatin 5 and LL-37, two antimicrobial peptides present in human saliva, on the functional and morphological properties of the Candida albicans cell membrane. Fluorescence microscopy and immunogold transmission electron microscopy revealed that LL-37 remained associated with the cell wall and cell membrane, whereas histatin 5 transmigrated over the membrane and accumulated intracellularly. Freeze-fracture electron microscopy revealed that LL-37 severely affected the membrane morphology, resulting in the disintegration of the membrane bilayer into discrete vesicles, and an instantaneous efflux of small molecules such as ATP as well as larger molecules such as proteins with molecular masses up to 40 kDa. The effects of histatin 5 on the membrane morphology were less pronounced, but still resulted in the efflux of nucleotides. As the morphological defects induced by histatin 5 are much smaller than those induced by LL-37, but the efflux of nucleotides is similar at comparable candidacidal concentrations, we suggest that the loss of nucleotides plays an important role in the killing process. PMID:15707390

Microtubules in Plant Cells: Strategies and Methods for Immunofluorescence, Transmission Electron Microscopy and Live Cell Imaging

PubMed Central

Celler, Katherine; Fujita, Miki; Kawamura, Eiko; Ambrose, Chris; Herburger, Klaus; Wasteneys, Geoffrey O.

2016-01-01

Microtubules are required throughout plant development for a wide variety of processes, and different strategies have evolved to visualize and analyze them. This chapter provides specific methods that can be used to analyze microtubule organization and dynamic properties in plant systems and summarizes the advantages and limitations for each technique. We outline basic methods for preparing samples for immunofluorescence labelling, including an enzyme-based permeabilization method, and a freeze-shattering method, which generates microfractures in the cell wall to provide antibodies access to cells in cuticle-laden aerial organs such as leaves. We discuss current options for live cell imaging of MTs with fluorescently tagged proteins (FPs), and provide chemical fixation, high pressure freezing/freeze substitution, and post-fixation staining protocols for preserving MTs for transmission electron microscopy and tomography. PMID:26498784

The detection of Giardia muris and Giardia lamblia cysts by immunofluorescence in animal tissues and fecal samples subjected to cycles of freezing and thawing.

PubMed

Erlandsen, S L; Sherlock, L A; Bemrick, W J

1990-04-01

The effects of freezing and thawing on the detection of selected Giardia spp. cysts were investigated using immunofluorescence, bright field microscopy, and low voltage scanning electron microscopy (SEM). Giardia muris cysts were obtained from either animal carcasses, fecal pellets, or isolated cyst preparations, whereas Giardia lamblia cysts were isolated from fecal samples. These samples were stained using an immunofluorescence technique after 1-3 freezing (-16 C) and thawing (20 C) cycles. Cysts were detected successfully by immunofluorescence in all samples. However, in those samples subjected to freeze-thawing, the cyst walls often became distorted and then were not detectable by bright field microscopy. Low voltage SEM demonstrated that the filaments in the distorted cyst wall underwent rearrangements of interfilament spacing. Quantitation of cyst recovery after freezing and thawing demonstrated that a substantial loss occurred after 1 cycle of alternating temperature when low concentrations of cysts were used, but not with high concentrations of cysts. Cyst recovery, after 3 freezing and thawing cycles, was dramatically lowered irrespective of the initial cyst concentration. These results demonstrated that immunofluorescence was an effective technique for the detection of Giardia spp. cysts in frozen samples and would suggest that freezing and thawing of fecal samples could prevent the detection of cysts when only bright field microscopy was employed.

Scanning electron microscopy fractography analysis of fractured hollow implants.

PubMed

Sbordone, Ludovico; Traini, Tonino; Caputi, Sergio; Scarano, Antonio; Bortolaia, Claudia; Piattelli, Adriano

2010-01-01

Fracture of the implant is one of the possible complications affecting dental implants; it is a rare event but of great clinical relevance. The aim of the present study was to perform a scanning electron microscopy (SEM) fractography evaluation of 7 International Team for oral Implantology (ITI) hollow implants removed because of fracture. The most common clinical risk factors, such as malocclusion, bruxism, and cantilevers on the prosthesis, were absent. Seven fractured ITI hollow implants were retrieved from 5 patients and were analyzed with the use of SEM. SEM analysis showed typical signs of a cleavage-type fracture. Fractures could be due to an association of multiple factors such as fatigue, inner defects, material electrochemical problems, and tensocorrosion.

Triblock copolyampholytes from 5-(N,N-dimethyl amino)isoprene styrene, and methacrylic acid: Synthesis and solution properties

NASA Astrophysics Data System (ADS)

Bieringer, R.; Abetz, V.; Müller, A. H. E.

ABC triblock copolymers of the type poly[5-(N,N-dimethyl amino)isoprene]-block-polystyrene-block-poly(tert-butyl methacrylate) (AiST) were synthesized and hydrolyzed to yield poly[5-(N,N-dimethyl amino)isoprene]-block-polystyrene-block-poly(methacrylic acid) (AiSA) triblock copolyampholytes. Due to a complex solubility behavior the solution properties of these materials had to be investigated in THF/water solvent mixtures. Potentiometric titrations of AiSA triblock copolyampholytes showed two inflection points with the A block being deprotonated prior to the Ai hydrochloride block thus forming a polyzwitterion at the isoelectric point (iep). The aggregation behavior was studied by dynamic light scattering (DLS) and freeze-fracture/transmission electron microscopy (TEM). Large vesicular structures with almost pH-independent radii were observed.

Nanosized cancer cell-targeted polymeric immunomicelles loaded with superparamagnetic iron oxide nanoparticles

NASA Astrophysics Data System (ADS)

Sawant, Rishikesh M.; Sawant, Rupa R.; Gultepe, Evin; Nagesha, Dattatri; Papahadjopoulos-Sternberg, Brigitte; Sridhar, Srinivas; Torchilin, Vladimir P.

2009-10-01

Stable 30-50 nm polymeric polyethylene glycol-phosphatidylethanolamine (PEG-PE)-based micelles entrapping superparamagnetic iron oxide nanoparticles (SPION) have been prepared. At similar concentrations of SPION, the SPION-micelles had significantly better magnetic resonance imaging (MRI) T2 relaxation signal compared to `plain' SPION. Freeze-fracture electron microscopy confirmed SPION entrapment in the lipid core of the PEG-PE micelles. To enhance the targeting capability of these micelles, their surface was modified with the cancer cell-specific anti-nucleosome monoclonal antibody 2C5 (mAb 2C5). Such mAb 2C5-SPION immunomicelles demonstrated specific binding with cancer cells in vitro and were able to bring more SPION to the cancer cells thus demonstrating the potential to be used as targeted MRI contrast agents for tumor imaging.

77 FR 12240 - Application(s) for Duty-Free Entry of Scientific Instruments

Federal Register 2010, 2011, 2012, 2013, 2014

2012-02-29

... Institute, 5353 Parkside Dr MC 19-RE, Jupiter, FL 33458. Instrument: Freeze Fracture/Freeze Etch device... localization of membrane proteins using freeze fracture replica immuno- gold labeling, including all kinds of receptors and channels. Because freeze-fracture replica immuno-gold labeling has a high sensitivity for the...

Correlative fractography: combining scanning electron microscopy and light microscopes for qualitative and quantitative analysis of fracture surfaces.

PubMed

Hein, Luis Rogerio de Oliveira; de Oliveira, José Alberto; de Campos, Kamila Amato

2013-04-01

Correlative fractography is a new expression proposed here to describe a new method for the association between scanning electron microscopy (SEM) and light microscopy (LM) for the qualitative and quantitative analysis of fracture surfaces. This article presents a new method involving the fusion of one elevation map obtained by extended depth from focus reconstruction from LM with exactly the same area by SEM and associated techniques, as X-ray mapping. The true topographic information is perfectly associated to local fracture mechanisms with this new technique, presented here as an alternative to stereo-pair reconstruction for the investigation of fractured components. The great advantage of this technique resides in the possibility of combining any imaging methods associated with LM and SEM for the same observed field from fracture surface.

Polyhedral protein cages encase synaptic vesicles and participate in their attachment to the active zone.

PubMed

Zampighi, G A; Fisher, R S

1997-08-01

In an effort to elucidate the interactions between synaptic vesicles and the membrane of the active zone, we have investigated the structure of interneuronal asymmetric synapses in the neocortex of adult rats using thin-sectioning, freeze-fracture, and negative staining electron microscopy. We identified three subtypes of spherical synaptic vesicles. Type I were agranular vesicles of 47.5 +/- 3.8 nm (mean SD, n = 24) in diameter usually seen aggregated in clusters in the presynaptic bouton. Type II synaptic vesicles were composed of a approximately 45-nm-diameter lipid bilayer sphere encased in a cage 77 +/- 4.6 nm (mean SD, n = 42) in diameter. The cage was composed of open-faced pentamers 20-22 nm/side arranged as a regular polyhedron. Type II caged vesicles were found in clusters at the boutons, adhered to the active zone, and were also present in axons. Type III synaptic vesicles appeared as electron-dense spheres 60-75 nm in diameter abutted to the membrane of the active zone. Clathrin-coated vesicles and pits of 116.6 +/- 9 nm (mean SD, n = 14) in diameter were also present in both the pre- and postsynaptic sides. Freeze-fracture showed that some intrinsic membrane proteins in the active zone were arranged as pentamers exhibiting the same dimension of those forming cages (approximately 22 nm/side). From these data, we concluded that: (a) the presynaptic bouton contains a heterogeneous population of "caged" and "plain" synaptic vesicles and (b) type II synaptic vesicles bind to receptors in the active zone. Therefore, current models of transmitter release should take into account the substantial heterogeneity of the vesicle population and the binding of vesicular cages to the membrane of the active zone.

Preservation of EDTA-expanded grid-mounted chromosomes and nuclei for electron microscopy using a specially designed freeze-dryer.

PubMed

Woods, P S; Ledbetter, M C; Tempel, N

1991-06-01

We describe methods for freezing and drying EDTA-expanded, fixed metaphase chromosomes and nuclei, attached to grids as whole-mounts, for transmission electron microscopy. These methods use a special apparatus that is simple to construct. While separate freezers and dryers are commercially available, one for freezing blocks of tissue by slamming them against a cold metal surface, and the other for vacuum drying the frozen tissue, our apparatus is designed for gentler, cryogenic liquid plunge freezing and drying, sequentially, in the same apparatus, thus avoiding any compression or damage to the specimen. Use of a cryoprotectant is not essential; however, good results are obtained more often when 20% ethanol is used. Freezing is accomplished by rapid propulsion of the grid, with specimens attached, into slushy N2 (-210 degrees C) within the drying chamber; drying is automatic, by either sublimation under vacuum or by solvent substitution using absolute ethanol followed by acetone, which, in turn, is removed with a critical-point dryer. The apparatus offers a means of drying chromosomes and nuclei in an expanded state, and avoids the shrinkage of these structures that occurs during stepwise passage through increasing concentrations of ethanol or acetone.

Animal-vegetal polarity in the plasma membrane of a molluscan egg: a quantitative freeze-fracture study.

PubMed

Speksnijder, J E; Mulder, M M; Dohmen, M R; Hage, W J; Bluemink, J G

1985-03-01

Using freeze-fracture electron microscopy, the numerical particle distribution in the fertilized Nassarius egg plasma membrane has been analyzed in four areas at different positions along the animal-vegetal axis of the egg. These areas can be distinguished by distinct microvilli patterns and differences in microvilli densities. In all areas, more IMPs (intramembrane particles) are present on the P face than on the corresponding E face. The ratio of the number of IMPs present on E and P face is similar in all areas (0.48-0.55) except for the most animal part of the vegetal hemisphere, where relatively more IMPs remain attached to the exterior half of the fractured membrane (E/P ratio = 0.88). The IMP density at the vegetal pole of the egg is considerably higher than in the animal hemisphere and in the animal part of the vegetal hemisphere. This difference is due to an increased number of IMPs in all size classes (4-18 nm). In the area adjacent to the vegetal pole the density of particles is also higher than in the two more animal areas, but here the difference is exclusively due to the smaller IMP size classes (4-8 nm). Statistical analysis of our data reveals that the area adjacent to the vegetal pole patch is significantly different from the other areas with respect to the distribution of the IMPs over the different IMP size classes. These results demonstrate the polar organization of the Nassarius egg plasma membrane. The possible role of this surface heterogeneity in the spatial organization of the egg cell and the later embryo is discussed.

Microstructure of Mixed Surfactant Solutions by Electron Microscopy

NASA Astrophysics Data System (ADS)

Naranjo, Edward

1995-01-01

Surfactant mixtures add a new dimension to the design of complex fluid microstructure. By combining different surfactants it is not only possible to modify aggregate morphology and control the macrascopic properties of colloidal dispersions but also to produce a variety of novel synergistic phases. Mixed systems produce new microstructures by altering the intermolecular and interaggregate forces in ways impossible for single component systems. In this dissertation, we report on the phase behavior and microstructure of several synthetic and biological surfactant mixtures as elucidated by freeze-fracture and cryo-transmission electron microscopy. We have discovered that stable, spontaneous unilamellar vesicles can be prepared from aqueous mixtures of commercially available single-tailed cationic and anionic surfactants. Vesicle stability is determined by the length and volume of the hydrocarbon chains of the "catanionic" pairs. Mixtures containing bulky or branched surfactant pairs (C _{16}/C_{12 -14}) in water produce defect-free fairly monodisperse equilibrium vesicles at high dilution. In contrast, mixtures of catanionic surfactants with highly asymmetric tails (C_{16}/C_8 ) form phases of porous vesicles, dilute lamellar L_{alpha}, and anomalous isotropic L_3 phases. Images of the microstructure by freeze-fracture microscopy show that the L_3 phase consists of multiconnected self-avoiding bilayers with saddle shaped curvature. The forces between bilayers of vesicle-forming cationic and anionic surfactant mixtures were also measured using the Surface Force Apparatus (SFA). We find that the vesicles are stabilized by a long range electrostatic repulsion at large separations (>20 A) and an additional salt-independent repulsive force below 20 A. The measured forces correlate very well with the ternary phase diagram and the vesicle microstructures observed by electron microscopy. In addition to studying ionic surfactants, we have also done original work with biological surfactants. We have found that subtle changes by surfactant additives to phosphatidylcholines (PC) produce dramatic changes in the microstructure of the composite that are impossible to determine from simple scattering experiments. Novel microstructures were observed at mole ratios from 4/1 to 9/1 long chain (Di-C_{16}PC)/short chain lipid (Di-C_7PC), including disc-like micelles and rippled bilayers at room temperature. We have also observed for the first time the formation of single layered ripple phase bilayer fragments. The formation of such fragments eliminates a number of theories of formation of this unique structure that depend on coupling between bilayers. In a similar system, dimyristoyl phosphatidylcholine (DMPC) mixed with the branched alcohol geraniol produces a bluish and extremely viscoelastic phase of giant multilamellar wormy vesicles. This phase shows the Weissenberg effect under flow due to the distortion of the entangled vesicles and may be related to fluid lamellar phases and L _3 phases often seen in surfactant-alcohol -water systems. Lysophosphatidylcholine, the single-chain counterpart of the diacyl phospholipids, can also form bilayer phases when combined with long-chain fatty acids in water. The phase transition characteristics and appearance of the bilayers in equimolar mixtures of lysolipid and fatty acid are similar to those of the diacyl-PC. Electron microscopy reveals large extended multilayers in mixtures with excess lysolipid and multilamellar vesicles in mixtures with excess fatty acid.

Xylem Surfactants Introduce a New Element to the Cohesion-Tension Theory1[OPEN

PubMed Central

Espino, Susana; Nima, Neda; Do, Aissa Y.T.; Michaud, Joseph M.; Papahadjopoulos-Sternberg, Brigitte; Yang, Jinlong; Steppe, Kathy

2017-01-01

Vascular plants transport water under negative pressure without constantly creating gas bubbles that would disable their hydraulic systems. Attempts to replicate this feat in artificial systems almost invariably result in bubble formation, except under highly controlled conditions with pure water and only hydrophilic surfaces present. In theory, conditions in the xylem should favor bubble nucleation even more: there are millions of conduits with at least some hydrophobic surfaces, and xylem sap is saturated or sometimes supersaturated with atmospheric gas and may contain surface-active molecules that can lower surface tension. So how do plants transport water under negative pressure? Here, we show that angiosperm xylem contains abundant hydrophobic surfaces as well as insoluble lipid surfactants, including phospholipids, and proteins, a composition similar to pulmonary surfactants. Lipid surfactants were found in xylem sap and as nanoparticles under transmission electron microscopy in pores of intervessel pit membranes and deposited on vessel wall surfaces. Nanoparticles observed in xylem sap via nanoparticle-tracking analysis included surfactant-coated nanobubbles when examined by freeze-fracture electron microscopy. Based on their fracture behavior, this technique is able to distinguish between dense-core particles, liquid-filled, bilayer-coated vesicles/liposomes, and gas-filled bubbles. Xylem surfactants showed strong surface activity that reduces surface tension to low values when concentrated as they are in pit membrane pores. We hypothesize that xylem surfactants support water transport under negative pressure as explained by the cohesion-tension theory by coating hydrophobic surfaces and nanobubbles, thereby keeping the latter below the critical size at which bubbles would expand to form embolisms. PMID:27927981

Xylem Surfactants Introduce a New Element to the Cohesion-Tension Theory.

PubMed

Schenk, H Jochen; Espino, Susana; Romo, David M; Nima, Neda; Do, Aissa Y T; Michaud, Joseph M; Papahadjopoulos-Sternberg, Brigitte; Yang, Jinlong; Zuo, Yi Y; Steppe, Kathy; Jansen, Steven

2017-02-01

Vascular plants transport water under negative pressure without constantly creating gas bubbles that would disable their hydraulic systems. Attempts to replicate this feat in artificial systems almost invariably result in bubble formation, except under highly controlled conditions with pure water and only hydrophilic surfaces present. In theory, conditions in the xylem should favor bubble nucleation even more: there are millions of conduits with at least some hydrophobic surfaces, and xylem sap is saturated or sometimes supersaturated with atmospheric gas and may contain surface-active molecules that can lower surface tension. So how do plants transport water under negative pressure? Here, we show that angiosperm xylem contains abundant hydrophobic surfaces as well as insoluble lipid surfactants, including phospholipids, and proteins, a composition similar to pulmonary surfactants. Lipid surfactants were found in xylem sap and as nanoparticles under transmission electron microscopy in pores of intervessel pit membranes and deposited on vessel wall surfaces. Nanoparticles observed in xylem sap via nanoparticle-tracking analysis included surfactant-coated nanobubbles when examined by freeze-fracture electron microscopy. Based on their fracture behavior, this technique is able to distinguish between dense-core particles, liquid-filled, bilayer-coated vesicles/liposomes, and gas-filled bubbles. Xylem surfactants showed strong surface activity that reduces surface tension to low values when concentrated as they are in pit membrane pores. We hypothesize that xylem surfactants support water transport under negative pressure as explained by the cohesion-tension theory by coating hydrophobic surfaces and nanobubbles, thereby keeping the latter below the critical size at which bubbles would expand to form embolisms. © 2017 American Society of Plant Biologists. All Rights Reserved.

Characterization of outer membranes isolated from Treponema pallidum, the syphilis spirochete.

PubMed

Radolf, J D; Robinson, E J; Bourell, K W; Akins, D R; Porcella, S F; Weigel, L M; Jones, J D; Norgard, M V

1995-11-01

Previous freeze-fracture electron microscopy (EM) studies have shown that the outer membrane (OM) of Treponema pallidum contains sparse transmembrane proteins. One strategy for molecular characterization of these rare OM proteins involves isolation of T. pallidum OMs. Here we describe a simple and extremely gentle method for OM isolation based upon isopycnic sucrose density gradient ultracentrifugation of treponemes following plasmolysis in 20% sucrose. Evidence that T. pallidum OMs were isolated included (i) the extremely low protein/lipid ratio of the putative OM fraction, (ii) a paucity of antigenic and/or biochemical markers for periplasmic, cytoplasmic membrane, and cytosolic compartments, and (iii) freeze-fracture EM demonstrating that the putative OMs contained intramembranous particles highly similar in size and density to those in native T. pallidum OMs. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed that the OMs contained a relatively small number of treponemal proteins, including several which did not appear to correspond to previously characterized T. pallidum antigens. Interestingly, these candidate rare OM proteins reacted poorly with syphilitic sera as determined by both conventional immunoblotting and enhanced chemiluminescence. Compared with whole cells, T. pallidum OMs were deficient in cardiolipin, the major lipoidal antigen reactive with antibodies in syphilitic sera. Also noteworthy was that other lipoidal constituents of OMs, including the recently discovered glycolipids, did not react with human syphilitic sera. These latter observations suggest that the poor antigenicity of virulent T. pallidum is a function of both the lipid composition and the low protein content of its OM.

A Global Approach for Quantitative Super Resolution and Electron Microscopy on Cryo and Epoxy Sections Using Self-labeling Protein Tags.

PubMed

Müller, Andreas; Neukam, Martin; Ivanova, Anna; Sönmez, Anke; Münster, Carla; Kretschmar, Susanne; Kalaidzidis, Yannis; Kurth, Thomas; Verbavatz, Jean-Marc; Solimena, Michele

2017-02-02

Correlative light and electron microscopy (CLEM) is a powerful approach to investigate the molecular ultrastructure of labeled cell compartments. However, quantitative CLEM studies are rare, mainly due to small sample sizes and the sensitivity of fluorescent proteins to strong fixatives and contrasting reagents for EM. Here, we show that fusion of a self-labeling protein to insulin allows for the quantification of age-distinct insulin granule pools in pancreatic beta cells by a combination of super resolution and transmission electron microscopy on Tokuyasu cryosections. In contrast to fluorescent proteins like GFP organic dyes covalently bound to self-labeling proteins retain their fluorescence also in epoxy resin following high pressure freezing and freeze substitution, or remarkably even after strong chemical fixation. This enables for the assessment of age-defined granule morphology and degradation. Finally, we demonstrate that this CLEM protocol is highly versatile, being suitable for single and dual fluorescent labeling and detection of different proteins with optimal ultrastructure preservation and contrast.

Cold-induced ultrastructural changes in bull and boar sperm plasma membranes.

PubMed

De Leeuw, F E; Chen, H C; Colenbrander, B; Verkleij, A J

1990-04-01

The effect of low temperatures on the ultrastructure of the plasma membrane of bull and boar spermatozoa was investigated. Cold-induced changes in the organization of sperm plasma membrane components were demonstrated by the use of fast-freezing combined with freeze-fracture electron microscopy. This preparation technique ensures fixation without artifacts. At 38 degrees C bull and boar spermatozoa exhibited a random distribution of intramembranous particles over the plasma membrane of both head and tail. Exposure to 0 degree C resulted in redistribution of the intramembranous particles: on the head and principal piece of bull spermatozoa and on the principal piece of boar spermatozoa, particle-free areas were observed, whereas on the boar sperm head, particle aggregates were present. The original particle distribution was restored upon rewarming of bull and boar spermatozoa to 38 degrees C, as well as after freezing and thawing of bull spermatozoa. Dilution of bull and boar semen into Tris-dilution buffer and Beltsville Thaw Solution-dilution buffer, respectively, could not prevent cold-induced redistribution of intramembranous particles. The observed particle reorganization upon cooling was interpreted as the result of lateral phase separation in the plasma membrane. Species-dependent differences in cold-induced ultrastructural changes were considered to be determined by lipid composition and asymmetry of the plasma membrane, and might be related to differences in cold resistance between species.

Aquaporin-0 Targets Interlocking Domains to Control the Integrity and Transparency of the Eye Lens

PubMed Central

Lo, Woo-Kuen; Biswas, Sondip K.; Brako, Lawrence; Shiels, Alan; Gu, Sumin; Jiang, Jean X.

2014-01-01

Purpose. Lens fiber cell membranes contain aquaporin-0 (AQP0), which constitutes approximately 50% of the total fiber cell membrane proteins and has a dual function as a water channel protein and an adhesion molecule. Fiber cell membranes also develop an elaborate interlocking system that is required for maintaining structural order, stability, and lens transparency. Herein, we used an AQP0-deficient mouse model to investigate an unconventional adhesion role of AQP0 in maintaining a normal structure of lens interlocking protrusions. Methods. The loss of AQP0 in AQP0−/− lens fibers was verified by Western blot and immunofluorescence analyses. Changes in membrane surface structures of wild-type and AQP0−/− lenses at age 3 to 12 weeks were examined with scanning electron microscopy. Preferential distribution of AQP0 in wild-type fiber cell membranes was analyzed with immunofluorescence and immunogold labeling using freeze-fracturing transmission electron microscopy. Results. Interlocking protrusions in young differentiating fiber cells developed normally but showed minor abnormalities at approximately 50 μm deep in the absence of AQP0 in all ages studied. Strikingly, protrusions in maturing fiber cells specifically underwent uncontrolled elongation, deformation, and fragmentation, while cells still retained their overall shape. Later in the process, these changes eventually resulted in fiber cell separation, breakdown, and cataract formation in the lens core. Immunolabeling at the light microscopy and transmission electron microscopy levels demonstrated that AQP0 was particularly enriched in interlocking protrusions in wild-type lenses. Conclusions. This study suggests that AQP0 exerts its primary adhesion or suppression role specifically to maintain the normal structure of interlocking protrusions that is critical to the integrity and transparency of the lens. PMID:24458158

Filling the gap: adding super-resolution to array tomography for correlated ultrastructural and molecular identification of electrical synapses at the C. elegans connectome.

PubMed

Markert, Sebastian Matthias; Britz, Sebastian; Proppert, Sven; Lang, Marietta; Witvliet, Daniel; Mulcahy, Ben; Sauer, Markus; Zhen, Mei; Bessereau, Jean-Louis; Stigloher, Christian

2016-10-01

Correlating molecular labeling at the ultrastructural level with high confidence remains challenging. Array tomography (AT) allows for a combination of fluorescence and electron microscopy (EM) to visualize subcellular protein localization on serial EM sections. Here, we describe an application for AT that combines near-native tissue preservation via high-pressure freezing and freeze substitution with super-resolution light microscopy and high-resolution scanning electron microscopy (SEM) analysis on the same section. We established protocols that combine SEM with structured illumination microscopy (SIM) and direct stochastic optical reconstruction microscopy (dSTORM). We devised a method for easy, precise, and unbiased correlation of EM images and super-resolution imaging data using endogenous cellular landmarks and freely available image processing software. We demonstrate that these methods allow us to identify and label gap junctions in Caenorhabditis elegans with precision and confidence, and imaging of even smaller structures is feasible. With the emergence of connectomics, these methods will allow us to fill in the gap-acquiring the correlated ultrastructural and molecular identity of electrical synapses.

Effect of microstructural anisotropy on fracture toughness of hot rolled 13Cr ODS steel - The role of primary and secondary cracking

NASA Astrophysics Data System (ADS)

Das, A.; Viehrig, H. W.; Bergner, F.; Heintze, C.; Altstadt, E.; Hoffmann, J.

2017-08-01

ODS steels have been known to exhibit anisotropic fracture behaviour and form secondary cracks. In this work, the factors responsible for the anisotropic fracture behaviour have been investigated using scanning electron microscopy and electron backscatter microscopy. Fracture toughness of hot rolled 13Cr ODS steel was determined using unloading compliance method for L-T and T-L orientations at various temperatures. L-T orientation had higher fracture toughness than T-L orientation and also contained more pronounced secondary cracking. Secondary cracks appeared at lower loads than primary cracks in both orientations. Primary crack propagation was found to be preferentially through fine grains in a bimodal microstructure. Grains were aligned and elongated the most towards rolling direction followed by T and S directions resulting in fracture anisotropy. Crystallographic texture and preferential alignment of Ti enriched particles parallel to rolling direction also contributed towards fracture anisotropy.

The control of ice crystal growth and effect on porous structure of konjac glucomannan-based aerogels.

PubMed

Ni, Xuewen; Ke, Fan; Xiao, Man; Wu, Kao; Kuang, Ying; Corke, Harold; Jiang, Fatang

2016-11-01

Konjac glucomannan (KGM)-based aerogels were prepared using a combination of sol-gel and freeze-drying methods. Preparation conditions were chosen to control ice crystal growth and aerogel structure formation. The ice crystals formed during pre-freezing were observed by low temperature polarizing microscopy, and images of aerogel pores were obtained by scanning electron microscopy. The size of ice crystals were calculated and size distribution maps were drawn, and similarly for aerogel pores. Results showed that ice crystal growth and aerogel pore sizes may be controlled by varying pre-freezing temperatures, KGM concentration and glyceryl monostearate concentration. The impact of pre-freezing temperatures on ice crystal growth was explained as combining ice crystal growth rate with nucleation rate, while the impacts of KGM and glyceryl monostearate concentration on ice crystal growth were interpreted based on their influences on sol network structure. Copyright © 2016 Elsevier B.V. All rights reserved.

Inactivation of Kudoa septempunctata in olive flounder meat by liquid freezing.

PubMed

Ohnishi, Takahiro; Akuzawa, Sayuri; Furusawa, Hiroko; Yoshinari, Tomoya; Kamata, Yoichi; Sugita-Konishi, Yoshiko

2014-01-01

Kudoa septempunctata in olive flounder meat was inactivated using 3 distinct freezing methods：liquid freezing for 5 min, air blast freezing at －30℃ for 5 h, and －80℃ for 1 h. The fracture curve of olive flounder meat subjected to liquid freezing resembled that of meat stored at 4℃, indicating that the structure of olive flounder muscle was well preserved. In contrast, air blast freezing induced the disappearance of the fracture point in the fracture curve, indicating that there was deterioration in the meat quality. Liquid freezing preserved the transparency of olive flounder meat to the same degree as that of meat stored at 4°C. However, air blast freezing induced meat cloudiness. These results indicate that liquid freezing can be used for K. septempunctata inactivation without affecting the meat quality.

Freeze-drying synthesis of three-dimensional porous LiFePO4 modified with well-dispersed nitrogen-doped carbon nanotubes for high-performance lithium-ion batteries

NASA Astrophysics Data System (ADS)

Tu, Xiaofeng; Zhou, Yingke; Song, Yijie

2017-04-01

The three-dimensional porous LiFePO4 modified with uniformly dispersed nitrogen-doped carbon nanotubes has been successfully prepared by a freeze-drying method. The morphology and structure of the porous composites are characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), and the electrochemical performances are evaluated using the constant current charge/discharge tests, cyclic voltammetry and electrochemical impedance spectroscopy. The nitrogen-doped carbon nanotubes are uniformly dispersed inside the porous LiFePO4 to construct a superior three-dimensional conductive network, which remarkably increases the electronic conductivity and accelerates the diffusion of lithium ion. The porous composite displays high specific capacity, good rate capability and excellent cycling stability, rendering it a promising positive electrode material for high-performance lithium-ion batteries.

Fracture resistance of dental nickel-titanium rotary instruments with novel surface treatment: Thin film metallic glass coating.

PubMed

Chi, Chih-Wen; Deng, Yu-Lun; Lee, Jyh-Wei; Lin, Chun-Pin

2017-05-01

Dental nickel-titanium (NiTi) rotary instruments are widely used in endodontic therapy because they are efficient with a higher success rate. However, an unpredictable fracture of instruments may happen due to the surface characteristics of imperfection (or irregularity). This study assessed whether a novel surface treatment could increase fatigue fracture resistance of dental NiTi rotary instruments. A 200- or 500-nm thick Ti-zirconium-boron (Ti-Zr-B) thin film metallic glass was deposited on ProTaper Universal F2 files using a physical vapor deposition process. The characteristics of coating were analyzed by scanning electron microscopy, transmission electron microscopy, and X-ray diffractometry. In cyclic fatigue tests, the files were performed in a simulated root canal (radius=5 mm, angulation=60°) under a rotating speed of 300rpm. The fatigue fractured cross sections of the files were analyzed with their fractographic performances through scanning electron microscopy images. The amorphous structure of the Ti-Zr-B coating was confirmed by transmission electron microscopy and X-ray diffractometry. The surface of treated files presented smooth morphologies without grinding irregularity. For the 200- and 500-nm surface treatment groups, the coated files exhibited higher resistance of cyclic fatigue than untreated files. In fractographic analysis, treated files showed significantly larger crack-initiation zone; however, no significant differences in the areas of fatigue propagation and catastrophic fracture were found compared to untreated files. The novel surface treatment of Ti-Zr-B thin film metallic glass on dental NiTi rotary files can effectively improve the fatigue fracture resistance by offering a smooth coated surface with amorphous microstructure. Copyright © 2016. Published by Elsevier B.V.

Aquaporin-Based Biomimetic Polymeric Membranes: Approaches and Challenges

PubMed Central

Habel, Joachim; Hansen, Michael; Kynde, Søren; Larsen, Nanna; Midtgaard, Søren Roi; Jensen, Grethe Vestergaard; Bomholt, Julie; Ogbonna, Anayo; Almdal, Kristoffer; Schulz, Alexander; Hélix-Nielsen, Claus

2015-01-01

In recent years, aquaporin biomimetic membranes (ABMs) for water separation have gained considerable interest. Although the first ABMs are commercially available, there are still many challenges associated with further ABM development. Here, we discuss the interplay of the main components of ABMs: aquaporin proteins (AQPs), block copolymers for AQP reconstitution, and polymer-based supporting structures. First, we briefly cover challenges and review recent developments in understanding the interplay between AQP and block copolymers. Second, we review some experimental characterization methods for investigating AQP incorporation including freeze-fracture transmission electron microscopy, fluorescence correlation spectroscopy, stopped-flow light scattering, and small-angle X-ray scattering. Third, we focus on recent efforts in embedding reconstituted AQPs in membrane designs that are based on conventional thin film interfacial polymerization techniques. Finally, we describe some new developments in interfacial polymerization using polyhedral oligomeric silsesquioxane cages for increasing the physical and chemical durability of thin film composite membranes. PMID:26264033

Niemann-Pick type C proteins promote microautophagy by expanding raft-like membrane domains in the yeast vacuole

PubMed Central

Tsuji, Takuma; Fujimoto, Megumi; Tatematsu, Tsuyako; Cheng, Jinglei; Orii, Minami; Takatori, Sho; Fujimoto, Toyoshi

2017-01-01

Niemann-Pick type C is a storage disease caused by dysfunction of NPC proteins, which transport cholesterol from the lumen of lysosomes to the limiting membrane of that compartment. Using freeze fracture electron microscopy, we show here that the yeast NPC orthologs, Ncr1p and Npc2p, are essential for formation and expansion of raft-like domains in the vacuolar (lysosome) membrane, both in stationary phase and in acute nitrogen starvation. Moreover, the expanded raft-like domains engulf lipid droplets by a microautophagic mechanism. We also found that the multivesicular body pathway plays a crucial role in microautophagy in acute nitrogen starvation by delivering sterol to the vacuole. These data show that NPC proteins promote microautophagy in stationary phase and under nitrogen starvation conditions, likely by increasing sterol in the limiting membrane of the vacuole. DOI: http://dx.doi.org/10.7554/eLife.25960.001 PMID:28590904

A comparison between rib fracture patterns in peri- and post-mortem compressive injury in a piglet model.

PubMed

Bradley, Amanda L; Swain, Michael V; Neil Waddell, J; Das, Raj; Athens, Josie; Kieser, Jules A

2014-05-01

Forensic biomechanics is increasingly being used to explain how observed injuries occur. We studied infant rib fractures from a biomechanical and morphological perspective using a porcine model. We used 24, 6th ribs of one day old domestic pigs Sus scrofa, divided into three groups, desiccated (representing post-mortem trauma), fresh ribs with intact periosteum (representing peri-mortem trauma) and those stored at -20°C. Two experiments were designed to study their biomechanical behaviour fracture morphology: ribs were axially compressed and subjected to four-point bending in an Instron 3339 fitted with custom jigs. Morphoscopic analysis of resultant fractures consisted of standard optical methods, micro-CT (μCT) and Scanning Electron Microscopy (SEM). During axial compression fresh ribs did not fracture because of energy absorption capabilities of their soft and fluidic components. In flexure tests, dry ribs showed typical elastic-brittle behaviour with long linear load-extension curves, followed by short non-linear elastic (hyperelastic) behaviour and brittle fracture. Fresh ribs showed initial linear-elastic behaviour, followed by strain softening and visco-plastic responses. During the course of loading, dry bone showed minimal observable damage prior to the onset of unstable fracture. Frozen then thawed bone showed similar patterns to fresh bone. Morphologically, fresh ribs showed extensive periosteal damage to the tensile surface with areas of collagen fibre pull-out along the tensile surface. While all dry ribs fractured precipitously, with associated fibre pull-out, the latter feature was absent in thawed ribs. Our study highlights the fact that under controlled loading, fresh piglet ribs (representing perimortem trauma) did not fracture through bone, but was associated with periosteal tearing. These results suggest firstly, that complete lateral rib fracture in infants may in fact not result from pure compression as has been previously assumed; and secondly, that freezing of bone during storage may affect its fracture behaviour. Copyright © 2013 Elsevier Ltd. All rights reserved.

In vitro early changes in intercellular junctions by treatment with a chemical carcinogen.

PubMed

Tachikawa, T; Kohno, Y; Matsui, Y; Yoshiki, S

1986-06-01

To examine early intercellular junction changes caused by treatment with 9,10-dimethyl-1,2-benzanthracene (DMBA), rat lingual epithelium was cultivated in isolation and observed by electrophysiological, freeze-fracture and whole-mount electron microscopy. Electrophysiological measurements showed a transient decrease in membrane potential of -10.2 mV 6 h after the treatment. It returned to almost the same level as that of the control group 1 day later. Six hours after treatment, input resistance decreased rapidly to 5.3 M omega but increased to 18.0 M omega 12 h after treatment. Transient reduction of input resistance and membrane potential occurred prior to the decrease in the coupling ratio 6 h after treatment with DMBA. In freeze-fracture replicas, the number of gap junctions decreased by approximately 45% of the control value 6 h after treatment with DMBA. At 12 h and thereafter, the number and area of gap junctions subsequently decreased by 60-80% of the control value. Alterations in the number and area of desmosomes were similar to those of the gap junctions. The formation of epithelial cytoskeletons, partially devoid of the 2-4 and 5-8 nm filaments was also observed. A decrease in the density of filament networks beneath the plasma membranes was especially apparent. Treatment with a carcinogen brought about morphological cellular changes as early as 6 h after treatment, and such early changes might trigger metabolic cellular abnormalities. Affected cells appear to move away from normal cells in a process of repeated destruction and revision of intercellular junctions, and cytoskeletons.

Raman microscopy of freeze-dried mouse eyeball-slice in conjunction with the "in vivo cryotechnique".

PubMed

Terada, Nobuo; Ohno, Nobuhiko; Saitoh, Sei; Fujii, Yasuhisa; Ohguro, Hiroshi; Ohno, Shinichi

2007-07-01

The wavelength of Raman-scattered light depends on the molecular composition of the substance. This is the first attempt to acquire Raman spectra of a mouse eyeball removed from a living mouse, in which the eyeball was preserved using the "in vivo cryotechnique" followed by freeze-drying. Eyeballs were cryofixed using a rapid freezing cryotechnique, and then sliced in the cryostat machine. The slices were sandwiched between glass slides, freeze-dried, and analyzed with confocal Raman microscopy. Important areas including various eyeball tissue layers were selected using bright-field microscopy, and then the Raman spectra were obtained at 240 locations. Four typical patterns of Raman spectra were electronically mapped on the specimen images obtained by the bright-field microscopy. Tissue organization was confirmed by embedding the same eyeball slice used for Raman spectra into epoxy resin and the thick sections were prepared with the inverted capsule method. Each Raman spectral pattern represents a different histological layer in the eyeball which was mapped by comparing the images of toluidine blue staining and Raman mapping with different colors. In the choroid and pigment cell layer, the Raman spectrum had two peaks, corresponding to melanin. Some of the peaks of the Raman spectra obtained from the blood vessels in sclera and the photoreceptor layer were similar to those obtained from the purified hemoglobin and rhodopsin proteins, respectively. Our experimental protocol can distinguish different tissue components with Raman microscopy; therefore, this method can be very useful for examining the distribution of a biological structures and/or chemical components in rapidly frozen freeze-dried tissue.

The dynamic failure behavior of tungsten heavy alloys subjected to transverse loads

NASA Astrophysics Data System (ADS)

Tarcza, Kenneth Robert

Tungsten heavy alloys (WHA), a category of particulate composites used in defense applications as kinetic energy penetrators, have been studied for many years. Even so, their dynamic failure behavior is not fully understood and cannot be predicted by numerical models presently in use. In this experimental investigation, a comprehensive understanding of the high-rate transverse-loading fracture behavior of WHA has been developed. Dynamic fracture events spanning a range of strain rates and loading conditions were created via mechanical testing and used to determine the influence of surface condition and microstructure on damage initiation, accumulation, and sample failure under different loading conditions. Using standard scanning electron microscopy metallographic and fractographic techniques, sample surface condition is shown to be extremely influential to the manner in which WHA fails, causing a fundamental change from externally to internally nucleated failures as surface condition is improved. Surface condition is characterized using electron microscopy and surface profilometry. Fracture surface analysis is conducted using electron microscopy, and linear elastic fracture mechanics is used to understand the influence of surface condition, specifically initial flaw size, on sample failure behavior. Loading conditions leading to failure are deduced from numerical modeling and experimental observation. The results highlight parameters and considerations critical to the understanding of dynamic WHA fracture and the development of dynamic WHA failure models.

New advances in scanning microscopy and its application to study parasitic protozoa.

PubMed

de Souza, Wanderley; Attias, Marcia

2018-07-01

Scanning electron microscopy has been used to observe and study parasitic protozoa for at least 40 years. However, field emission electron sources, as well as improvements in lenses and detectors, brought the resolution power of scanning electron microscopes (SEM) to a new level. Parallel to the refinement of instruments, protocols for preservation of the ultrastructure, immunolabeling, exposure of cytoskeleton and inner structures of parasites and host cells were developed. This review is focused on protozoan parasites of medical and veterinary relevance, e.g., Toxoplasma gondii, Tritrichomonas foetus, Giardia intestinalis, and Trypanosoma cruzi, compilating the main achievements in describing the fine ultrastructure of their surface, cytoskeleton and interaction with host cells. Two new resources, namely, Helium Ion Microscopy (HIM) and Slice and View, using either Focused Ion Beam (FIB) abrasion or Microtome Serial Sectioning (MSS) within the microscope chamber, combined to backscattered electron imaging of fixed (chemically or by quick freezing followed by freeze substitution and resin embedded samples is bringing an exponential amount of valuable information. In HIM there is no need of conductive coating and the depth of field is much higher than in any field emission SEM. As for FIB- and MSS-SEM, high resolution 3-D models of areas and volumes larger than any other technique allows can be obtained. The main results achieved with all these technological tools and some protocols for sample preparation are included in this review. In addition, we included some results obtained with environmental/low vacuum scanning microscopy and cryo-scanning electron microscopy, both promising, but not yet largely employed SEM modalities. Copyright © 2018. Published by Elsevier Inc.

Investigation of high temperature fracture of T-111 and ASTAR-811C

NASA Technical Reports Server (NTRS)

Gold, R. E.

1971-01-01

The high temperature deformation and fracture behavior of T-111 and ASTAR-811C were studied over the temperature range 982 to 2205 C (1800 to 4000 F). As-cast and wrought-recrystallized material as well as GTA welds in sheet and plate were evaluated using conventional tensile and creep tests. Post test examinations were performed using optical metallography, scanning electron microscopy and transmission electron microscopy. A high temperature region of reduced ductility, in terms of tensile elongation, was identified for both alloys. The reduction in tensile elongation became more severe with increase in grain size, being near catastrophic for the as-cast specimens. Optical and electron metallography indicated that even for failures at very low total strain, considerable deformation of a very localized nature had occurred prior to fracture.

Tandem High-pressure Freezing and Quick Freeze Substitution of Plant Tissues for Transmission Electron Microscopy

PubMed Central

Bobik, Krzysztof; Dunlap, John R.; Burch-Smith, Tessa M.

2014-01-01

Since the 1940s transmission electron microscopy (TEM) has been providing biologists with ultra-high resolution images of biological materials. Yet, because of laborious and time-consuming protocols that also demand experience in preparation of artifact-free samples, TEM is not considered a user-friendly technique. Traditional sample preparation for TEM used chemical fixatives to preserve cellular structures. High-pressure freezing is the cryofixation of biological samples under high pressures to produce very fast cooling rates, thereby restricting ice formation, which is detrimental to the integrity of cellular ultrastructure. High-pressure freezing and freeze substitution are currently the methods of choice for producing the highest quality morphology in resin sections for TEM. These methods minimize the artifacts normally associated with conventional processing for TEM of thin sections. After cryofixation the frozen water in the sample is replaced with liquid organic solvent at low temperatures, a process called freeze substitution. Freeze substitution is typically carried out over several days in dedicated, costly equipment. A recent innovation allows the process to be completed in three hours, instead of the usual two days. This is typically followed by several more days of sample preparation that includes infiltration and embedding in epoxy resins before sectioning. Here we present a protocol combining high-pressure freezing and quick freeze substitution that enables plant sample fixation to be accomplished within hours. The protocol can readily be adapted for working with other tissues or organisms. Plant tissues are of special concern because of the presence of aerated spaces and water-filled vacuoles that impede ice-free freezing of water. In addition, the process of chemical fixation is especially long in plants due to cell walls impeding the penetration of the chemicals to deep within the tissues. Plant tissues are therefore particularly challenging, but this protocol is reliable and produces samples of the highest quality. PMID:25350384

Fine structures and ion images on fresh frozen dried ultrathin sections by transmission electron and scanning ion microscopy

NASA Astrophysics Data System (ADS)

Takaya, K.; Okabe, M.; Sawataishi, M.; Takashima, H.; Yoshida, T.

2003-01-01

Ion microscopy (IM) of air-dried or freeze-dried cryostat and semi-thin cryosections has provided ion images of elements and organic substances in wide areas of the tissue. For reproducible ion images by a shorter time of exposure to the primary ion beam, fresh frozen dried ultrathin sections were prepared by freezing the tissue in propane chilled with liquid nitrogen, cryocut at 60 nm, mounted on grids and silicon wafer pieces, and freeze-dried. Rat Cowper gland and sciatic nerve, bone marrow of the rat administered of lithium carbonate, tree frog and African toad spleen and buffy coat of atopic dermatitis patients were examined. Fine structures and ion images of the corresponding areas in the same or neighboring sections were observed by transmission electron microscopy (TEM) followed by sector type and time-of-flight type IM. Cells in the buffy coat contained larger amounts of potassium and magnesium while plasma had larger amounts of sodium and calcium. However, in the tissues, lithium, sodium, magnesium, calcium and potassium were distributed in the cell and calcium showed a granular appearance. A granular cell of the tree frog spleen contained sodium and potassium over the cell and magnesium and calcium were confined to granules.

Freezing without Ice Crystal Damage: Semithin and Ultrathin Frozen Sections of Ethanol-Infiltrated Tissue for Microscopy, with Applications to Immunocytochemistry

NASA Astrophysics Data System (ADS)

Christensen, A. Kent; Lowry, Terry B.

1995-10-01

Ethanol (ethyl alcohol) has long been a standard reagent used in preparing tissues for light and electron microscopy. After fixation, tissues are usually dehydrated with ethanol before being embedded in paraffin or plastic. In this study we show that the ethanol-infiltrated tissue can be frozen and sectioned directly without embedding. When tissue impregnated with ethanol is cooled below about [minus sign]117°C with liquid nitrogen, the ethanol solidifies without appreciable crystallization. The frozen tissue can then be sectioned in a commercial cryoultramicrotome that is set at [minus sign]155 to [minus sign]170°C to produce semithin frozen sections (0.5 to 3 [mu]m thick) for light microscopy or ultrathin frozen sections (50 to 100 nm thick) for electron microscopy. Sections are picked up and mounted on glass slides or EM grids by means that are in current use for ice ultrathin frozen sectioning. Because there is no apparent freezing damage, the morphology in these ethanol frozen sections of unembedded tissue appears generally quite good, often resembling that obtained by conventional EM techniques. Examples are provided that illustrate the use of this material for immunocytochemistry at the light and electron microscope levels.

Improvement of the fracture toughness of hydroxyapatite (HAp) by incorporation of carboxyl functionalized single walled carbon nanotubes (CfSWCNTs) and nylon

DOE PAGES

Khanal, Suraj P.; Mahfuz, Hassan; Rondinone, Adam Justin; ...

2015-11-12

The potential of improving the fracture toughness of synthetic hydroxyapatite (HAp) by incorporating carboxyl functionalized single walled carbon nanotubes (CfSWCNTs) and polymerized ε-caprolactam (nylon) was researched. A series of HAp samples with CfSWCNTs concentrations varying from 0 to 1.5 wt.%, without, and with nylon addition was prepared. X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM) were used to characterize the samples. The three point bending test was applied to measure the fracture toughness of the composites. A reproducible value of 3.6 ± 0.3 MPa.√m was found for samples containing 1 wt.% CfSWCNTs and nylon. This valuemore » is in the range of the cortical bone fracture toughness. Lastly, the increase of the CfSWCNTs content results to decrease of the fracture toughness, and formation of secondary phases.« less

Freeze drying-assisted synthesis of Pt@reduced graphene oxide nanocomposites as excellent hydrogen sensor

NASA Astrophysics Data System (ADS)

Lu, Xiaojing; Song, Xinjie; Gu, Cuiping; Ren, Haibo; Sun, Yufeng; Huang, Jiarui

2018-05-01

Quick and efficient detection of low concentrations of hydrogen remains a challenge because of the stability of hydrogen. A sensor based on reduced oxide graphene functionalized with Pt nanoparticles is successfully fabricated using a freeze-drying method followed by heat treatment. The structure and morphology of the Pt@rGO nanocomposites are well analyzed by X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, Raman spectroscopy, and X-ray photoelectron spectroscopy. The as-prepared Pt@rGO nanocomposites show excellent hydrogen gas sensing properties at a low working temperature of 50 °C. The sensitivity toward 0.5% hydrogen is 8%. The response and recovery times of the sensor exposed to 0.5% hydrogen are 63 and 104 s, respectively. The gas-sensing mechanism of Pt@rGO sensor is also discussed.

Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast Saccharomyces cerevisiae.

PubMed

Fišerová, Jindřiška; Richardson, Christine; Goldberg, Martin W

2016-01-01

Immunolabeling electron microscopy is a challenging technique with demands for perfect ultrastructural and antigen preservation. High-pressure freezing offers an excellent way to fix cellular structure. However, its use for immunolabeling has remained limited because of the low frequency of labeling due to loss of protein antigenicity or accessibility. Here we present a protocol for immunogold labeling of the yeast Saccharomyces cerevisiae that gives specific and multiple labeling while keeping the finest structural details. We use the protocol to reveal the organization of individual nuclear pore complex proteins and the position of transport factors in the yeast Saccharomyces cerevisiae in relation to actual transport events.

Analysis of rolling fracture of the conticasted and tandem rolled blanks of low alloyed aluminum

NASA Astrophysics Data System (ADS)

Li, Yong; Zeng, Lingping; Jiao Xie, Xian

2018-01-01

Optical microscopy, electron microscopy and energy spectrum were used to test the morphology of grains, as-cast microstructure and secondary phases in confiscated and tandem rolled planks of 8011 low alloying aluminum alloy. It can be concluded that the existence of inhomogeneous secondary FeSiAl phases lead to the fracture of planks during rolling.

Reversible Cryopreservation of Living Cells Using an Electron Microscopy Cryo-Fixation Method.

PubMed

Huebinger, Jan; Han, Hong-Mei; Grabenbauer, Markus

2016-01-01

Rapid cooling of aqueous solutions is a useful approach for two important biological applications: (I) cryopreservation of cells and tissues for long-term storage, and (II) cryofixation for ultrastructural investigations by electron and cryo-electron microscopy. Usually, both approaches are very different in methodology. Here we show that a novel, fast and easy to use cryofixation technique called self-pressurized rapid freezing (SPRF) is-after some adaptations-also a useful and versatile technique for cryopreservation. Sealed metal tubes with high thermal diffusivity containing the samples are plunged into liquid cryogen. Internal pressure builds up reducing ice crystal formation and therefore supporting reversible cryopreservation through vitrification of cells. After rapid rewarming of pressurized samples, viability rates of > 90% can be reached, comparable to best-performing of the established rapid cooling devices tested. In addition, the small SPRF tubes allow for space-saving sample storage and the sealed containers prevent contamination from or into the cryogen during freezing, storage, or thawing.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khanal, Suraj P.; Mahfuz, Hassan; Rondinone, Adam Justin

The potential of improving the fracture toughness of synthetic hydroxyapatite (HAp) by incorporating carboxyl functionalized single walled carbon nanotubes (CfSWCNTs) and polymerized ε-caprolactam (nylon) was researched. A series of HAp samples with CfSWCNTs concentrations varying from 0 to 1.5 wt.%, without, and with nylon addition was prepared. X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM) were used to characterize the samples. The three point bending test was applied to measure the fracture toughness of the composites. A reproducible value of 3.6 ± 0.3 MPa.√m was found for samples containing 1 wt.% CfSWCNTs and nylon. This valuemore » is in the range of the cortical bone fracture toughness. Lastly, the increase of the CfSWCNTs content results to decrease of the fracture toughness, and formation of secondary phases.« less

Effect of Solvent Dielectric Constant on the Formation of Large Flat Bilayer Stacks in a Lecithin/Hexadecanol Hydrogel.

PubMed

Nakagawa, Yasuharu; Nakazawa, Hiromitsu; Kato, Satoru

2016-07-12

We investigated the effect of dielectric properties of the aqueous medium on the novel type of hydrogel composed of a crude lecithin mixture (PC70) and hexadecanol (HD), in which charged sheet-like bilayers are kept far apart due to interbilayer repulsive interaction. We used dipropylene glycol (DPG) as a modifier of the dielectric properties and examined its effect on the hydrogel by synchrotron X-ray diffraction, differential scanning calorimetry (DSC), polarized optical microscopy, and freeze-fracture electron microscopy. We found that at a DPG weight fraction in the aqueous medium WDPG ≈ 0.4, the bilayer organization is transformed into unusually large flat bilayer stacks with a regular lamellar spacing of 6.25 nm and consequently disintegration of the hydrogel takes place. Semiquantitative calculation of the interbilayer interaction energy based on the Deyaguin-Landau-Verwey-Overbeek (DLVO) theory suggested that the reduction of the aqueous medium dielectric constant ε by DPG may lower the energy barrier preventing flat bilayers from coming closer together. We inferred that the size of the bilayer sheet increases because the reduction of ε promotes protonation of acidic lipids that work as edge-capping molecules.

Corrigendum to 'On the influence of microstructure on the fracture behaviour of hot extruded ferritic ODS steels' [J. Nucl. Mater. 497 (2017) 60-75

NASA Astrophysics Data System (ADS)

Das, A.; Viehrig, H. W.; Altstadt, E.; Heintze, C.; Hoffmann, J.

2018-02-01

ODS steels are known to show inferior fracture properties as compared to ferritic martensitic non-ODS steels. Hot extruded 13Cr ODS steel however, showed excellent fracture toughness at a temperature range from room temperature to 400 °C. In this work, the factors which resulted in superior and anisotropic fracture behaviour were investigated by comparing different orientations of two hot extruded materials using scanning electron, electron backscatter and transmission electron microscopy. Fracture behaviour of the two materials was compared using unloading compliance fracture toughness tests. Anisotropic fracture toughness was predominantly influenced by grain morphology. Superior fracture toughness in 13Cr ODS-KIT was predominantly influenced by factors such as smaller void inducing particle size and higher sub-micron particle-matrix interfacial strength.

Crystal Initiation Structures in Developing Enamel: Possible Implications for Caries Dissolution of Enamel Crystals

PubMed Central

Robinson, Colin; Connell, Simon D.

2017-01-01

Investigations of developing enamel crystals using Atomic and Chemical Force Microscopy (AFM, CFM) have revealed a subunit structure. Subunits were seen in height images as collinear swellings about 30 nM in diameter on crystal surfaces. In friction mode they were visible as positive regions. These were similar in size (30–50 nM) to collinear spherical structures, presumably mineral matrix complexes, seen in developing enamel using a freeze fracturing/freeze etching procedure. More detailed AFM studies on mature enamel suggested that the 30–50 nM structures were composed of smaller units, ~10–15 nM in diameter. These were clustered in hexagonal or perhaps a spiral arrangement. It was suggested that these could be the imprints of initiation sites for mineral precipitation. The investigation aimed at examining original freeze etched images at high resolution to see if the smaller subunits observed using AFM in mature enamel were also present in developing enamel i.e., before loss of the organic matrix. The method used was freeze etching. Briefly samples of developing rat enamel were rapidly frozen, fractured under vacuum, and ice sublimed from the fractured surface. The fractured surface was shadowed with platinum or gold and the metal replica subjected to high resolution TEM. For AFM studies high-resolution tapping mode imaging of human mature enamel sections was performed in air under ambient conditions at a point midway between the cusp and the cervical margin. Both AFM and freeze etch studies showed structures 30–50 nM in diameter. AFM indicated that these may be clusters of somewhat smaller structures ~10–15 nM maybe hexagonally or spirally arranged. High resolution freeze etching images of very early enamel showed ~30–50 nM spherical structures in a disordered arrangement. No smaller units at 10–15 nM were clearly seen. However, when linear arrangements of 30–50 nM units were visible the picture was more complex but also smaller units including ~10–15 nM units could be observed. Conclusions: Structures ~10–15 nM in diameter were detected in developing enamel. While the appearance was complex, these were most evident when the 30–5 nM structures were in linear arrays. Formation of linear arrays of subunits may be associated with the development of mineral initiation sites and attendant processing of matrix proteins. PMID:28670283

State transitions and physicochemical aspects of cryoprotection and stabilization in freeze-drying of Lactobacillus rhamnosus GG (LGG).

PubMed

Pehkonen, K S; Roos, Y H; Miao, S; Ross, R P; Stanton, C

2008-06-01

The frozen and dehydrated state transitions of lactose and trehalose were determined and studied as factors affecting the stability of probiotic bacteria to understand physicochemical aspects of protection against freezing and dehydration of probiotic cultures. Lactobacillus rhamnosus GG was frozen (-22 or -43 degrees C), freeze-dried and stored under controlled water vapour pressure (0%, 11%, 23% and 33% relative vapour pressure) conditions. Lactose, trehalose and their mixture (1 : 1) were used as protective media. These systems were confirmed to exhibit relatively similar state transition and water plasticization behaviour in freeze-concentrated and dehydrated states as determined by differential scanning calorimetry. Ice formation and dehydrated materials were studied using cold-stage microscopy and scanning electron microscopy. Trehalose and lactose-trehalose gave the most effective protection of cell viability as observed from colony forming units after freezing, dehydration and storage. Enhanced cell viability was observed when the freezing temperature was -43 degrees C. State transitions of protective media affect ice formation and cell viability in freeze-drying and storage. Formation of a maximally freeze-concentrated matrix with entrapped microbial cells is essential in freezing prior to freeze-drying. Freeze-drying must retain a solid amorphous state of protectant matrices. Freeze-dried matrices contain cells entrapped in the protective matrices in the freezing process. The retention of viability during storage seems to be controlled by water plasticization of the protectant matrix and possibly interactions of water with the dehydrated cells. Highest cell viability was obtained in glassy protective media. This study shows that physicochemical properties of protective media affect the stability of dehydrated cultures. Trehalose and lactose may be used in combination, which is particularly important for the stabilization of probiotic bacteria in dairy systems.

Nanoscale patterning of STIM1 and Orai1 during store-operated Ca2+ entry.

PubMed

Perni, Stefano; Dynes, Joseph L; Yeromin, Andriy V; Cahalan, Michael D; Franzini-Armstrong, Clara

2015-10-06

Stromal interacting molecule (STIM) and Orai proteins constitute the core machinery of store-operated calcium entry. We used transmission and freeze-fracture electron microscopy to visualize STIM1 and Orai1 at endoplasmic reticulum (ER)-plasma membrane (PM) junctions in HEK 293 cells. Compared with control cells, thin sections of STIM1-transfected cells possessed far more ER elements, which took the form of complex stackable cisternae and labyrinthine structures adjoining the PM at junctional couplings (JCs). JC formation required STIM1 expression but not store depletion, induced here by thapsigargin (TG). Extended molecules, indicative of STIM1, decorated the cytoplasmic surface of ER, bridged a 12-nm ER-PM gap, and showed clear rearrangement into small clusters following TG treatment. Freeze-fracture replicas of the PM of Orai1-transfected cells showed extensive domains packed with characteristic "particles"; TG treatment led to aggregation of these particles into sharply delimited "puncta" positioned upon raised membrane subdomains. The size and spacing of Orai1 channels were consistent with the Orai crystal structure, and stoichiometry was unchanged by store depletion, coexpression with STIM1, or an Orai1 mutation (L273D) affecting STIM1 association. Although the arrangement of Orai1 channels in puncta was substantially unstructured, a portion of channels were spaced at ∼15 nm. Monte Carlo analysis supported a nonrandom distribution for a portion of channels spaced at ∼15 nm. These images offer dramatic, direct views of STIM1 aggregation and Orai1 clustering in store-depleted cells and provide evidence for the interaction of a single Orai1 channel with small clusters of STIM1 molecules.

Effects of freezing and thawing on texture, microstructure and cell wall composition changes in papaya tissues.

PubMed

Phothiset, Suphatta; Charoenrein, Sanguansri

2014-01-30

During storage, frozen fruit may be thawed and refrozen many times before consumption, which may be extremely damaging to the texture of the frozen fruit and reverse the advantage of fast freezing. The effects of freezing and thawing on texture, microstructure and cell wall composition changes in papaya tissues were investigated. The frozen-thawed papayas had an increase in drip loss and a decrease in firmness with increasing number of freeze-thaw cycles. Light microscopy showed irregular shapes and cell damage in parenchyma cells of frozen-thawed papayas, whereas transmission electron microscopy showed loss of cell wall materials in middle lamella. Moreover, destruction of cell wall was observed after being subjected to five freeze-thaw cycles. These changes related with a significant decrease in alcohol-insoluble solids, Na₂CO₃- and 24% KOH-soluble fractions and an increase in the water-, EDTA- and 4% KOH-soluble fractions. This was due to a decrease in the molecular mass of pectic and hemicellulosic polymers in frozen-thawed papayas using high-performance size-exclusion chromatography. The freezing and thawing processes caused fine structural damage and cell wall composition changes which contributed to a loss of drip volume and firmness of papaya tissues. © 2013 Society of Chemical Industry.

Microstructure and mechanical properties of quenched and tempered 300M steel

NASA Technical Reports Server (NTRS)

Youngblood, J. L.; Raghavan, M.

1978-01-01

Type 300M steel, which is being used for the landing gear on the space shuttle orbiter, was subjected to a wide range of quenched and tempered heat treatments. The plane-strain fracture toughness and the tensile ultimate and yield strengths were evaluated. Cryogenic mechanical properties were obtained for conventionally heat-treated steel. The microstructure of all heat-treated test coupons was studied both optically and by transmission electron microscopy. Fracture surfaces were studied by means of scanning electron microscopy. Results indicate that substantial improvement in toughness with no loss in strength can be accomplished in quenched and tempered steel by austenitizing at 1255 K or higher. Low fracture toughness in conventionally austenitized 300M steel (1144 K) appears to be caused by undissolved precipitates, seen both in the submicrostructure and on the fracture surface, which promote failure by quasi-cleavage. The precipitates appeared to dissolve in the range 1200 to 1255 K.

Fractographic examination of racing greyhound central (navicular) tarsal bone failure surfaces using scanning electron microscopy.

PubMed

Tomlin, J L; Lawes, T J; Blunn, G W; Goodship, A E; Muir, P

2000-09-01

The greyhound is a fatigue fracture model of a short distance running athlete. Greyhounds have a high incidence of central (navicular) tarsal bone (CTB) fractures, which are not associated with overt trauma. We wished to determine whether these fractures occur because of accumulation of fatigue microdamage. We hypothesized that bone from racing dogs would show site-specific microdamage accumulation, causing predisposition to structural failure. We performed a fractographic examination of failure surfaces from fractured bones using scanning electron microscopy and assessed microcracking observed at the failure surface using a visual analog scale. Branching arrays of microcracks were seen in failure surfaces of CTB and adjacent tarsal bones, suggestive of compressive fatigue failure. Branching arrays of microcracks were particularly prevalent in remodeled trabecular bone that had become compact. CTB fractures showed increased microdamage when compared with other in vivo fractures (adjacent tarsal bone and long bone fractures), and ex vivo tarsal fractures induced by monotonic loading (P < 0.02). It was concluded that greyhound racing and training often results in CTB structural failure, because of accumulation and coalescence of branching arrays of fatigue microcracks, the formation of which appears to be predisposed to adapted bone.

The ability of the Antarctic nematode Panagrolaimus davidi to survive intracellular freezing is dependent upon nutritional status.

PubMed

Raymond, Mélianie R; Wharton, David A

2013-02-01

The Antarctic nematode Panagrolaimus davidi is the best documented example of an animal surviving intracellular freezing and the only animal so far shown to survive such freezing throughout its tissues. However, a recent study found that after exposure to a freezing stress that produced intracellular freezing in a proportion of nematodes, the resulting survival levels could be explained if those nematodes that froze intracellularly had died. We have thus re-examined the survival of intracellular freezing in this nematode. The ability to survive a freezing exposure that is likely to produce intracellular freezing (freezing at -10 °C) declines with culture age. In cultures that are fed regularly, the ability to survive freezing at -10 °C increases, but in starved cultures freezing survival declines. Survival of intracellular freezing in fed cultures was confirmed using cryomicroscopy, staining of cells with vital dyes and by freeze substitution and transmission electron microscopy. We have thus confirmed that P. davidi can survive intracellular freezing and shown that this ability is dependent upon them being well fed. The effect of culture conditions on the nutrient status of the nematodes should thus be an important factor in the design of experiments.

Effects of freezing, freeze drying and convective drying on in vitro gastric digestion of apples.

PubMed

Dalmau, Maria Esperanza; Bornhorst, Gail M; Eim, Valeria; Rosselló, Carmen; Simal, Susana

2017-01-15

The influence of processing (freezing at -196°C in liquid N2, FN sample; freeze-drying at -50°C and 30Pa, FD sample; and convective drying at 60°C and 2m/s, CD sample) on apple (var. Granny Smith) behavior during in vitro gastric digestion was investigated. Dried apples (FD and CD samples) were rehydrated prior to digestion. Changes in carbohydrate composition, moisture, soluble solids, acidity, total polyphenol content (TPC), and antioxidant activity (AA) of apple samples were measured at different times during digestion. Processing resulted in disruption of the cellular structure during digestion, as observed by scanning electron microscopy, light microscopy, and changes in carbohydrate composition. Moisture content increased (6-11% dmo), while soluble solids (55-78% dmo), acidity (44-72% dmo), total polyphenol content (30-61% dmo), and antioxidant activity (41-87%) decreased in all samples after digestion. Mathematical models (Weibull and exponential models) were used to better evaluate the influence of processing on apple behavior during gastric digestion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Inter-wall bridging induced peeling of multi-walled carbon nanotubes during tensile failure in aluminum matrix composites.

PubMed

Chen, Biao; Li, Shufeng; Imai, Hisashi; Umeda, Junko; Takahashi, Makoto; Kondoh, Katsuyoshi

2015-02-01

In situ scanning electron microscopy (SEM) observation of a tensile test was performed to investigate the fracturing behavior of multi-walled carbon nanotubes (MWCNTs) in powder metallurgy Al matrix composites. A multiple peeling phenomenon during MWCNT fracturing was clearly observed. Its formation mechanism and resultant effect on the composite strength were examined. Through transition electron microscopy characterizations, it was observed that defective structures like inter-wall bridges cross-linked adjacent walls of MWCNTs. This structure was helpful to improve the inter-wall bonding conditions, leading to the effective load transfer between walls and resultant peeling behaviors of MWCNTs. These results might provide new understandings of the fracturing mechanisms of carbon nanotube reinforcements for designing high-performance nanocomposites. Copyright © 2014 Elsevier Ltd. All rights reserved.

Osmotically Induced Reversible Transitions in Lipid-DNA Mesophases

PubMed Central

Danino, Dganit; Kesselman, Ellina; Saper, Gadiel; Petrache, Horia I.; Harries, Daniel

2009-01-01

We follow the effect of osmotic pressure on isoelectric complexes that self-assemble from mixtures of DNA and mixed neutral and cationic lipids. Using small angle x-ray diffraction and freeze-fracture cryo-electron microscopy, we find that lamellar complexes known to form in aqueous solutions can reversibly transition to hexagonal mesophases under high enough osmotic stress exerted by adding a neutral polymer. Using molecular spacings derived from x-ray diffraction, we estimate the reversible osmotic pressure-volume (Π-V) work needed to induce this transition. We find that the transition free energy is comparable to the work required to elastically bend lipid layers around DNA. Consistent with this, the required work is significantly lowered by an addition of hexanol, which is known to soften lipid bilayers. Our findings not only help to resolve the free-energy contributions associated with lipid-DNA complex formation, but they also demonstrate the importance that osmotic stress can have to the macromolecular phase geometry in realistic biological environments. PMID:19348739

Size Effects on Deformation and Fracture of Scandium Deuteride Films.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Teresi, C. S.; Hintsala, E.; Adams, David P.

Metal hydride films have been observed to crack during production and use, prompting mechanical property studies of scandium deuteride films. The following focuses on elastic modulus, fracture, and size effects observed in the system for future film mechanical behavior modeling efforts. Scandium deuteride films were produced through the deuterium charging of electron beam evaporated scandium films using X-ray diffraction, scanning Auger microscopy, and electron backscatter diffraction to monitor changes in the films before and after charging. Scanning electron microscopy, nanoindentation, and focused ion beam machined micropillar compression tests were used for mechanical characterization of the scandium deuteride films. The micropillarsmore » showed a size effect for flow stress, indicating that film thickness is a relevant tuning parameter for film performance, and that fracture was controlled by the presence of grain boundaries. Elastic modulus was determined by both micropillar compression and nanoindentation to be approximately 150 GPa, Fracture studies of bulk film channel cracking as well as compression induced cracks in some of the pillars yielded a fracture toughness around 1.0 MPa-m1/2. Preliminary Weibull distributions of fracture in the micropillars are provided. Despite this relatively low value of fracture toughness, scandium deuteride micropillars can undergo a large degree of plasticity in small volumes and can harden to some degree, demonstrating the ductile and brittle nature of this material« less

Effect of Nb on Delayed Fracture Resistance of Ultra-High Strength Martensitic Steels

NASA Astrophysics Data System (ADS)

Song, Rongjie; Fonstein, Nina; Pottore, Narayan; Jun, Hyun Jo; Bhattacharya, Debanshu; Jansto, Steve

Ultra-high strength steels are materials of considerable interest for automotive and structural applications and are increasingly being used in those areas. Higher strength, however, makes steels more prone to hydrogen embrittlement (HE). The effects of Nb and other alloying elements on the hydrogen-induced delayed fracture resistance of cold rolled martensitic steels with ultra-high strength 2000 MPa were studied using an acid immersion test, thermal desorption analysis (TDA) and measuring of permeation. The microstructure was characterized by high resolution field emission Scanning Electron Microscopy (SEM) with Electron Backscattered Diffraction (EBSD) and Transmission Electron Microscopy (TEM). It was shown that the combined addition of Nb significantly improved the delayed fracture resistance of investigated steel. The addition of Nb to alloyed martensitic steels resulted in very apparent grain refinement of the prior austenite grain size. The Nb microalloyed steel contained a lower diffusible hydrogen content during thermal desorption analysis as compared to the base steel and had a higher trapped hydrogen amount after charging. The reason that Nb improved the delayed fracture resistance of steels can be attributed mostly to both hydrogen trapping and grain refinement.

A pore-scale study of fracture dynamics in rock using X-ray micro-CT under ambient freeze-thaw cycling.

PubMed

De Kock, Tim; Boone, Marijn A; De Schryver, Thomas; Van Stappen, Jeroen; Derluyn, Hannelore; Masschaele, Bert; De Schutter, Geert; Cnudde, Veerle

2015-03-03

Freeze-thaw cycling stresses many environments which include porous media such as soil, rock and concrete. Climate change can expose new regions and subject others to a changing freeze-thaw frequency. Therefore, understanding and predicting the effect of freeze-thaw cycles is important in environmental science, the built environment and cultural heritage preservation. In this paper, we explore the possibilities of state-of-the-art micro-CT in studying the pore scale dynamics related to freezing and thawing. The experiments show the development of a fracture network in a porous limestone when cooling to -9.7 °C, at which an exothermal temperature peak is a proxy for ice crystallization. The dynamics of the fracture network are visualized with a time frame of 80 s. Theoretical assumptions predict that crystallization in these experiments occurs in pores of 6-20.1 nm under transient conditions. Here, the crystallization-induced stress exceeds rock strength when the local crystal fraction in the pores is 4.3%. The location of fractures is strongly related to preferential water uptake paths and rock texture, which are visually identified. Laboratory, continuous X-ray micro-CT scanning opens new perspectives for the pore-scale study of ice crystallization in porous media as well as for environmental processes related to freeze-thaw fracturing.

[Production and characteristics of bacteria-labeled talc dust for experimental air hygiene studies].

PubMed

Ohgke, H; Oldenburg, B; Gropengiesser, R; Herbst, M

1983-04-01

Freeze-drying of suspensions of Micrococcus luteus together with talc yields bacteria-labelled dust. This material can be used in experimental air hygiene. Loss of viability due to drying in air during experiments can be expected to be negligible. A wide range of particle diameters (1 to greater than 23 micron) is available. Scanning electron microscopy shows the bacteria sticking on talc particles after freeze-drying (Fig. 3a + b). Viable counts of the material decreased very slowly on storage.

Electron microscopy of Drosophila garland cell nephrocytes: Optimal preparation, immunostaining and STEM tomography.

PubMed

Hochapfel, Florian; Denk, Lucia; Maaßen, Christine; Zaytseva, Yulia; Rachel, Reinhard; Witzgall, Ralph; Krahn, Michael P

2018-01-29

Due to its structural and molecular similarities to mammalian podocytes, the Drosophila nephrocyte emerged as a model system to study podocyte development and associated diseases. Similar to podocytes, nephrocytes establish a slit diaphragm between foot process-like structures in order to filter the hemolymph. One major obstacle in nephrocyte research is the distinct visualization of this subcellular structure to assess its integrity. Therefore, we developed a specialized dissection and fixation protocol, including high pressure freezing and freeze substitution techniques, to improve the preservation of the intricate ultrastructural details necessary for electron microscopic assessment. By means of scanning transmission electron microscopy (STEM) tomography, a three-dimensional dataset was generated to further understand the complex architecture of the nephrocyte channel system. Moreover, a staining protocol for immunolabeling of ultrathin sections of Epon-embedded nephrocytes is discussed, which allows the reliable detection of GFP-tagged fusion proteins combined with superior sample preservation. Due to the growing number of available GFP-trap fly lines, this approach is widely applicable for high resolution localization studies in wild type and mutant nephrocytes. © 2018 Wiley Periodicals, Inc.

Evaluation of poly (vinyl alcohol) based cryogel-zinc oxide nanocomposites for possible applications as wound dressing materials.

PubMed

Chaturvedi, Archana; Bajpai, Anil K; Bajpai, Jaya; K Singh, Sunil

2016-08-01

In this investigation cryogels composed of poly (vinyl alcohol) (PVA) were prepared by repeated freeze thaw method followed by in situ precipitation of zinc oxide nanoparticles within the cryogel networks. Fourier transformed infrared spectroscopy (FTIR), scanning electron microscopy (SEM), transmission electron microscopy (TEM), and X-ray diffraction (XRD), Energy dispersive X-ray spectroscopy (EDX) were used to characterize the nanocomposites. The morphologies of native PVA cryogels and PVA cryogel-ZnO nanocomposites were observed by scanning electron microscopy (SEM), transmission electron microscopy (TEM) techniques. The SEM analysis suggested that cryogels show a well-defined porous morphology whereas TEM micrographs revealed the presence of nearly spherical and well separated zinc oxide nanoparticles with diameter<100nm. XRD results showed all relevant Bragg's reflections for crystal structure of zinc oxide nanoparticles. Thermo gravimetric-differential thermal analysis (TG-DTA) was conducted to evaluate thermal stability of the nanocomposites. Mechanical properties of nanocomposites were determined in terms of tensile strength and percent elongation. Biocompatible nature was ascertained by anti-haemolytic activity, bovine serum albumin (blood protein) adsorption and in vitro cytotoxicity tests. The prepared nanocomposites were also investigated for swelling and deswelling behaviours. The results revealed that both the swelling and deswelling process depend on the chemical composition of the nanocomposites, number of freeze-thaw cycles, pH and temperature of the swelling medium. The developed biocompatible PVA cryogel-ZnO nanocomposites were also tested for antibacterial activities against both Gram-negative and Gram-positive bacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

ON THE FREEZING AND IDENTIFICATION OF LIPID MONOLAYER 2-D ARRAYS FOR CRYOELECTRON MICROSCOPY

PubMed Central

Taylor, Dianne W.; Kelly, Deborah F.; Cheng, Anchi; Taylor, Kenneth A.

2008-01-01

Lipid monolayers provide a convenient vehicle for the crystallization of biological macromolecules for 3-D electron microscopy. Although numerous examples of 3-D images from 2-D protein arrays have been described from negatively stained specimens, only six structures have been done from frozen hydrated specimens. We describe here a method that makes high quality frozen-hydrated specimens of lipid monolayer arrays for cryoelectron microscopy. The method uses holey carbon films with patterned holes for monolayer recovery, blotting and plunge freezing to produce thin aqueous films which cover >90% of the available grid area. With this method, even specimens with relatively infrequent crystals can be screened using automated data collection techniques. Though developed for microscopic examination of 2-D arrays, the method may have wider application to the preparation of single particle specimens for 3-D image reconstruction. PMID:17561414

Effects of stripped oil shale retort water on fishes, birds, and mammals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nystrom, R.R.

1983-01-01

Golden hamsters (Mesocricetus auratus Water), coturnix quail (Coturnix coturnix Teminck and Schlegal), fathead minnows (Pimphales promelas Rafinesque), and rainbow trout (Salmo gairdneri Richardson) were subjected to various exposures of stripped oil shale retort water (SRW). Chronic low-level exposures of all experimental animals to SRW revealed no adverse histological effects attributable to SRW. Also, production and development of second generation fathead minnows and coturnix quail exposed to SRW was normal. Subacute exposure of rainbow trout to SRW produced ultrastructural changes detected by transmission, scanning, and freeze fracture electron microscopy) in the gill, liver, and kidney tissues. The gills showed a swellingmore » of secondary lamellae, disorganization of normal tissue architecture, and sloughing of respiratory cells. The liver contained lamellar bodies not seen in the controls. Relatively large, electron dense, membrane-bounded deposits were present in proximal tubule cells of the kidney. Sodium arsenite (a significant component of SRW) was shown to cause swelling of granular endosplasmic reticulum in quail liver tissue with an acute exposure. This effect could be related to the fact that arsenic inhibits ATP production, which would decrease the ability of the sodium pumps to maintain a normal osmotic balance.« less

Correlative light-electron fractography for fatigue striations characterization in metallic alloys.

PubMed

Hein, Luis Rogerio de Oliveira; de Oliveira, José Alberto; de Campos, Kamila Amato

2013-09-01

The correlative light-electron fractography technique combines correlative microscopy concepts to the extended depth-from-focus reconstruction method, associating the reliable topographic information of 3-D maps from light microscopy ordered Z-stacks to the finest lateral resolution and large focus depth from scanning electron microscopy. Fatigue striations spacing analysis can be precisely measured, by correcting the mean surface tilting with the knowledge of local elevation data from elevation maps. This new technique aims to improve the accuracy of quantitative fractography in fatigue fracture investigations. Copyright © 2013 Wiley Periodicals, Inc.

The effects of alumina nanofillers on mechanical properties of high-performance epoxy resin.

PubMed

Zhang, Hui; Zhang, Hui; Tang, Longcheng; Liu, Gang; Zhang, Daijun; Zhou, Lingyun; Zhang, Zhong

2010-11-01

In the past decade extensive studies have been focused on mechanical properties of inorganic nanofiller/epoxy matrices. In this work we systematically investigated the mechanical properties of nano-alumina-filled E-54/4, 4-diaminodiphenylsulphone (DDS) epoxy resins, which were prepared via combining high-speed mixing with three-roll milling. Homogeneous dispersion of nano-alumina with small agglomerates was obtained in epoxy resin, which was confirmed using transmission electron microscopy (TEM). The static/dynamic modulus, tensile strength and fracture toughness of the nanocomposites were found to be simultaneously enhanced with addition of nano-alumina fillers. About 50% and 80% increases of K(IC) and G(IC) were achieved in nanocomposite filled with 18.4 wt% alumina nanofillers, as compared to that of the unfilled epoxy resin. Furthermore, the corresponding fracture surfaces of tensile and compact tension samples were examined by scanning electron microscopy (SEM) and atomic force microscopy (AFM) techniques in order to identify the relevant fracture mechanisms involved. Various fracture features including cavities/debonding of nanofiller, local plastic deformation as well as crack pinning/deflection were found to be operative in the presence of nano-alumina fillers.

Cryo-scanning electron microscopy investigation of the Octopus Vulgaris arm structures for the design of an octopus-like arm artefact.

PubMed

Minnocci, Antonio; Cianchetti, Matteo; Mazzolai, Barbara; Sebastiani, Luca; Laschi, Cecilia

2015-12-01

Octopus vulgaris is a cephalopod of the Octopodidae family. It has four pairs of arms and two rows of suckers which perform many functions, including bending and elongation. For this reason the octopus was chosen as model to develop a new generation of soft-body robots. In order to explain some of the fine structures of the octopus arm in relation to its specific ability, we examined the external and internal structures of O. vulgaris arms in a frozen-hydrated state using cryo-scanning electron microscopy. The arms showed skin with a very complex design that is useful to elongation, and a pore pattern distribution on their surface which is functional to cutaneous oxygen uptake. The analysis of freeze-fractured frozen-hydrated arm samples allowed us to describe the developmental differences in the relative proportion of the areas of axial nerve cord, intrinsic and extrinsic musculature, in relation to the growth of the arms and of the increase in functional capability. In the suckers, we analyzed the shedding mechanisms in the outer part of the infundibulum and described the outer and inner characteristics of the denticles, showing in detail their pore system, which is fundamental for their ability to explore the environment. These results are discussed by considering their possible application in the design of new octopus-like artefacts, which will be able to take advantage of some of these ultrastructure characteristics and achieve advanced bioinspired functionalities. © 2015 Wiley Periodicals, Inc.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brummett, A.R.; Dumont, J.N.

A comparative study of the chorions of eggs of northern and southern populations of Fundulus heteroclitus by scanning and transmission electron microscopy reveals striking differences. Chorionic fibrils of eggs of the northern (Woods Hole) population are very long, approx. 1.5 ..mu.. in diameter, and very sparsely distributed; the chorionic surface between attached fibrils is dotted with small protuberances. Most of the fibrils of the eggs of a southern (South Carolina) population are shorter, approx. 0.5 ..mu.. in diameter, and very densely distributed. The South Carolina eggs have a few longer and thicker (approx. 1.0 ..mu..) fibrils in the vicinity ofmore » the micropyle. The fibrils of the Woods Hole eggs are club-shaped at their bases, surrounded by a collar of ''jelly'' at their attachment points, and are seated in an indentation in the chorion. Those of the South Carolina eggs show no such basal modifications and appear to extend from a small chorionic hillock. A surface coat of jelly is present on the ovulated eggs of both populations but appears to be thicker and denser on the eggs of southern origin. Scanning electron microscopy of freeze-fractured preparations of ovarian tissue from the two populations shows that the chorionic fibrils are present and attached to the developing chorion as soon as it is visible. Jelly is not present on the surface on the unovulated eggs. The data are discussed from the standpoint of considerations of the taxonomy and distribution of the species, and questions are raised concerning the possible significance of the structural differences observed.« less

Self-assembled multicompartment liquid crystalline lipid carriers for protein, peptide, and nucleic acid drug delivery.

PubMed

Angelova, Angelina; Angelov, Borislav; Mutafchieva, Rada; Lesieur, Sylviane; Couvreur, Patrick

2011-02-15

Lipids and lipopolymers self-assembled into biocompatible nano- and mesostructured functional materials offer many potential applications in medicine and diagnostics. In this Account, we demonstrate how high-resolution structural investigations of bicontinuous cubic templates made from lyotropic thermosensitive liquid-crystalline (LC) materials have initiated the development of innovative lipidopolymeric self-assembled nanocarriers. Such structures have tunable nanochannel sizes, morphologies, and hierarchical inner organizations and provide potential vehicles for the predictable loading and release of therapeutic proteins, peptides, or nucleic acids. This Account shows that structural studies of swelling of bicontinuous cubic lipid/water phases are essential for overcoming the nanoscale constraints for encapsulation of large therapeutic molecules in multicompartment lipid carriers. For the systems described here, we have employed time-resolved small-angle X-ray scattering (SAXS) and high-resolution freeze-fracture electronic microscopy (FF-EM) to study the morphology and the dynamic topological transitions of these nanostructured multicomponent amphiphilic assemblies. Quasi-elastic light scattering and circular dichroism spectroscopy can provide additional information at the nanoscale about the behavior of lipid/protein self-assemblies under conditions that approximate physiological hydration. We wanted to generalize these findings to control the stability and the hydration of the water nanochannels in liquid-crystalline lipid nanovehicles and confine therapeutic biomolecules within these structures. Therefore we analyzed the influence of amphiphilic and soluble additives (e.g. poly(ethylene glycol)monooleate (MO-PEG), octyl glucoside (OG), proteins) on the nanochannels' size in a diamond (D)-type bicontinuous cubic phase of the lipid glycerol monooleate (MO). At body temperature, we can stabilize long-living swollen states, corresponding to a diamond cubic phase with large water channels. Time-resolved X-ray diffraction (XRD) scans allowed us to detect metastable intermediate and coexisting structures and monitor the temperature-induced phase sequences of mixed systems containing glycerol monooleate, a soluble protein macromolecule, and an interfacial curvature modulating agent. These observed states correspond to the stages of the growth of the nanofluidic channel network. With the application of a thermal stimulus, the system becomes progressively more ordered into a double-diamond cubic lattice formed by a bicontinuous lipid membrane. High-resolution freeze-fracture electronic microscopy indicates that nanodomains are induced by the inclusion of proteins into nanopockets of the supramolecular cubosomic assemblies. These results contribute to the understanding of the structure and dynamics of functionalized self-assembled lipid nanosystems during stimuli-triggered LC phase transformations.

Torsion fracture of carbon nanocoils

NASA Astrophysics Data System (ADS)

Yonemura, Taiichiro; Suda, Yoshiyuki; Tanoue, Hideto; Takikawa, Hirofumi; Ue, Hitoshi; Shimizu, Kazuki; Umeda, Yoshito

2012-10-01

We fix a carbon nanocoil (CNC) on a substrate in a focused ion beam instrument and then fracture the CNC with a tensile load. Using the CNC spring index, we estimate the maximum to average stress ratio on the fractured surface to range from 1.3 to 1.7, indicating stress concentration on the coil wire inner edge. Scanning electron microscopy confirms a hollow region on the inner edge of all fractured surfaces.

High-quality ultrastructural preservation using cryofixation for 3D electron microscopy of genetically labeled tissues

PubMed Central

Boassa, Daniela; Hu, Junru; Romoli, Benedetto; Phan, Sebastien; Dulcis, Davide

2018-01-01

Electron microscopy (EM) offers unparalleled power to study cell substructures at the nanoscale. Cryofixation by high-pressure freezing offers optimal morphological preservation, as it captures cellular structures instantaneously in their near-native state. However, the applicability of cryofixation is limited by its incompatibility with diaminobenzidine labeling using genetic EM tags and the high-contrast en bloc staining required for serial block-face scanning electron microscopy (SBEM). In addition, it is challenging to perform correlated light and electron microscopy (CLEM) with cryofixed samples. Consequently, these powerful methods cannot be applied to address questions requiring optimal morphological preservation. Here, we developed an approach that overcomes these limitations; it enables genetically labeled, cryofixed samples to be characterized with SBEM and 3D CLEM. Our approach is broadly applicable, as demonstrated in cultured cells, Drosophila olfactory organ and mouse brain. This optimization exploits the potential of cryofixation, allowing for quality ultrastructural preservation for diverse EM applications. PMID:29749931

Ultrafast endocytosis at mouse hippocampal synapses

NASA Astrophysics Data System (ADS)

Watanabe, Shigeki; Rost, Benjamin R.; Camacho-Pérez, Marcial; Davis, M. Wayne; Söhl-Kielczynski, Berit; Rosenmund, Christian; Jorgensen, Erik M.

2013-12-01

To sustain neurotransmission, synaptic vesicles and their associated proteins must be recycled locally at synapses. Synaptic vesicles are thought to be regenerated approximately 20s after fusion by the assembly of clathrin scaffolds or in approximately 1s by the reversal of fusion pores via `kiss-and-run' endocytosis. Here we use optogenetics to stimulate cultured hippocampal neurons with a single stimulus, rapidly freeze them after fixed intervals and examine the ultrastructure using electron microscopy--`flash-and-freeze' electron microscopy. Docked vesicles fuse and collapse into the membrane within 30ms of the stimulus. Compensatory endocytosis occurs within 50 to 100ms at sites flanking the active zone. Invagination is blocked by inhibition of actin polymerization, and scission is blocked by inhibiting dynamin. Because intact synaptic vesicles are not recovered, this form of recycling is not compatible with kiss-and-run endocytosis; moreover, it is 200-fold faster than clathrin-mediated endocytosis. It is likely that `ultrafast endocytosis' is specialized to restore the surface area of the membrane rapidly.

Effects of Cryogenic Temperature on Fracture Toughness of Core-Shell Rubber (CSR) Toughened Epoxy Nanocomposites

NASA Technical Reports Server (NTRS)

Wang, J.; Cannon, S. A.; Magee, D.; Schneider, J. A.

2008-01-01

This study investigated the effects of core-shell rubber (CSR) nanoparticles on the mechanical properties and fracture toughness of an epoxy resin at ambient and liquid nitrogen (LN2) temperatures. Varying amounts of Kane Ace MX130 toughening agent were added to a commercially available EPON 862/Epikure W epoxy resin. Elastic modulus was calculated using quasi-static tensile data. Fracture toughness was evaluated by the resulting breaking energy measured in Charpy impact tests conducted on an instrumented drop tower. The size and distribution of the CSR nanoparticles were characterized using Transmission Electron Microscopy (TEM) and Small Angle X-ray Scattering (SAXS). Scanning Electron Microscopy (SEM) was used to study the fracture surface morphology. The addition of the CSR nanoparticles increased the breaking energy with negligible change in elastic modulus and ultimate tensile stress (UTS). At ambient temperature the breaking energy increased with increasing additions of the CSR nanoparticles, while at LN2 temperatures, it reached a maximum at 5 wt% CSR concentration. KEY WORDS: liquid nitrogen (LN2) properties, fracture toughness, core-shell rubber (CSR).

Some personal and historical notes on the utility of "deep-etch" electron microscopy for making cell structure/function correlations.

PubMed

Heuser, John E

2014-11-01

This brief essay talks up the advantages of metal replicas for electron microscopy and explains why they are still the best way to image frozen cells in the electron microscope. Then it explains our approach to freezing, namely the Van Harreveld trick of "slamming" living cells onto a supercold block of metal sprayed with liquid helium at -269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment. This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most "true to life" views of cells out of them with a minimum of hassle. Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the "founding fathers" that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are "irresistible and half transparent … their meaning buried under only a few years of work," and "reasonable working hypotheses are already suggested by the ultrastructural organization itself." © 2014 Heuser.

Freeze-drying of silica nanoparticles: redispersibility toward nanomedicine applications.

PubMed

Picco, Agustin S; Ferreira, Larissa F; Liberato, Michelle S; Mondo, Gabriela B; Cardoso, Mateus B

2018-01-01

To study freeze-drying of silica nanoparticles (SiO 2 NPs) in order to find suitable conditions to produce lyophilized powders with no aggregation after resuspension and storage. SiO 2 NPs were synthesized using a Stöber-based procedure, and characterized by scanning electron microscopy, dynamic light scattering and nitrogen adsorption/desorption isotherms. SiO 2 NPs hydrodynamic diameters were compared prior and after freeze-drying in the presence/absence of carbohydrate protectants. Glucose was found to be the most suitable protectant against the detrimental effects of lyophilization. The minimum concentration of carbohydrate required to effectively protect SiO 2 NPs from aggregation during freeze-drying is influenced by the nanoparticle's size and texture. Negligible aggregation was observed during storage. Carbohydrates can be used during SiO 2 NPs freeze-drying process to obtain redispersable solids that maintain original sizes without residual aggregation.

Some Structural Observations of Self-Assembling, Fibrillar Gels Composed of Two-Directional Bolaform Arborols

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, J.

2005-01-12

Arborols are dumbbell shaped molecules (bolaform amphiphiles) in which a hydrophobic spacer separates two hydrophilic end groups. They are a valuable model for naturally occurring fibers, such as actin or amyloid. Applications to materials science can be envisioned. On cooling from warm aqueous or methanolic solutions, arborols spontaneously assemble into long fibers. When the solutions are above a certain concentration that depends on the hydrophilic/hydrophobic balance, this leads to thermally reversible gels stabilized by a mechanism that is poorly understood. With the help of wide angle X-ray scattering, details of the arborol fiber and gel structure were obtained on wetmore » gels. The characteristic dimensions of the fibers vary in a sensible fashion with the molecular specifics. Solvent character appears to affect the average domain length of arborols stacked into fibers. Fluorescently labeled arborols were prepared. The label does not prevent incorporation into the fibrillar structure, rendering fibril bundles visible in wet gels. Bundles are visible in concentrated gels, but not in less concentrated sols. These results are consistent with observations of dried arborols using atomic force microscopy and with previously published freeze-fracture electron microscopy and small angle X-ray scattering experiments on dried gels.« less

Failure analysis of explanted sternal wires.

PubMed

Shih, Chun-Ming; Su, Yea-Yang; Lin, Shing-Jong; Shih, Chun-Che

2005-05-01

To classify and understand the mechanisms of surface damages and fracture mechanisms of sternal wires, explanted stainless steel sternal wires were collected from patients with sternal dehiscence following open-heart surgery. Surface alterations and fractured ends of sternal wires were examined and analyzed. Eighty fractured wires extracted from 25 patients from January 1999 to December 2003, with mean implantation interval of 55+/-149 days (range 5-729 days) after cardiac surgery, were studied by various techniques. The extracted wires were cleaned and the fibrotic tissues were removed. Irregularities and fractured ends were assayed by a scanning electron microscopy. After stereomicroscopy and documentation, the explants were cleaned with 1% sodium hypochlorite to remove the blood and tissues and was followed by cleaned with deionized water and alcohol. The explants were examined by stereomicroscopy, and irregularities on surface and fracture surfaces of sternal wires were assayed by scanning electron microscopy, energy dispersive X-ray analysis (EDAX) and X-ray mapping. The explants with surrounding fibrotic tissue were stained and examined with stereomicroscopy and transmission electronic microscopy. Corrosion pits were found on the surface of explanted sternal wires. EDAX and X-ray mapping examinations revealed diminution of nickel concentration in the severely corroded pits on sternal wires. A feature of transgranular cracking was observed for stress corrosion cracking and striation character for typical corrosion fatigue was also identified. TEM examination of tissue showed the metallic particles in phagolysosomes of macrophages inside the surrounding sternal tissue. The synergic effect of hostile environment and the stress could be the precursors of failures for sternal wires.

A Freeze Substitution Fixation-Based Gold Enlarging Technique for EM Studies of Endocytosed Nanogold-Labeled Molecules

PubMed Central

He, Wanzhong; Kivork, Christine; Machinani, Suman; Morphew, Mary K.; Gail, Anna M.; Tesar, Devin B.; Tiangco, Noreen E.; McIntosh, J. Richard; Bjorkman, Pamela J.

2007-01-01

We have developed methods to locate individual ligands that can be used for electron microscopy studies of dynamic events during endocytosis and subsequent intracellular trafficking. The methods are based on enlargement of 1.4 nm Nanogold attached to an endocytosed ligand. Nanogold, a small label that does not induce misdirection of ligand-receptor complexes, is ideal for labeling ligands endocytosed by live cells, but is too small to be routinely located in cells by electron microscopy. Traditional pre-embedding enhancement protocols to enlarge Nanogold are not compatible with high pressure freezing/freeze substitution fixation (HPF/FSF), the most accurate method to preserve ultrastructure and dynamic events during trafficking. We have developed an improved enhancement procedure for chemically-fixed samples that reduced autonucleation, and a new pre-embedding gold-enlarging technique for HPF/FSF samples that preserved contrast and ultrastructure and can be used for high-resolution tomography. We evaluated our methods using labeled Fc as a ligand for the neonatal Fc receptor. Attachment of Nanogold to Fc did not interfere with receptor binding or uptake, and gold-labeled Fc could be specifically enlarged to allow identification in 2D projections and in tomograms. These methods should be broadly applicable to many endocytosis and transcytosis studies. PMID:17723309

Applications of synchrotron x-ray diffraction topography to fractography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bilello, J.C.

1983-01-01

Fractographs have been taken using a variety of probes each of which produces different types of information. Methods which have been used to examine fracture surfaces include: (a) optical microscopy, particularly interference contrast methods, (b) scanning electron microscopy (SEM), (c) SEM with electron channelling, (d) SEM with selected-area electron channelling, (e) Berg-Barrett (B-B) topography, and now (f) synchrotron x-radiation fractography (SXRF). This review concentrated on the role that x-ray methods can play in such studies. In particular, the ability to nondestructively assess the subsurface microstructure associated with the fracture to depths of the order of 5 to 10 ..mu..m becomesmore » an important attribute for observations of a large class of semi-brittle metals, semiconductors and ceramics.« less

Comparative Proteomic and Morphological Change Analyses of Staphylococcus aureus During Resuscitation From Prolonged Freezing

PubMed Central

Suo, Biao; Yang, Hua; Wang, Yuexia; Lv, Haipeng; Li, Zhen; Xu, Chao; Ai, Zhilu

2018-01-01

When frozen, Staphylococcus aureus survives in a sublethally injured state. However, S. aureus can recover at a suitable temperature, which poses a threat to food safety. To elucidate the resuscitation mechanism of freezing survived S. aureus, we used cells stored at -18°C for 90 days as controls. After resuscitating the survived cells at 37°C, the viable cell numbers were determined on tryptic soy agar with 0.6% yeast extract (TSAYE), and the non-injured-cell numbers were determined on TSAYE supplemented with 10% NaCl. The results showed that the total viable cell number did not increase within the first 3 h of resuscitation, but the osmotic regulation ability of freezing survived cells gradually recovered to the level of healthy cells, which was evidenced by the lack of difference between the two samples seen by differential cell enumeration. Scanning electron microscopy (SEM) showed that, compared to late exponential stage cells, some frozen survived cells underwent splitting and cell lysis due to deep distortion and membrane rupture. Transmission electron microscopy (TEM) showed that, in most of the frozen survived cells, the nucleoids (low electronic density area) were loose, and the cytoplasmic matrices (high electronic density area) were sparse. Additionally, a gap was seen to form between the cytoplasmic membranes and the cell walls in the frozen survived cells. The morphological changes were restored when the survived cells were resuscitated at 37°C. We also analyzed the differential proteome after resuscitation using non-labeled high-performance liquid chromatography–mass spectrometry (HPLC-MS). The results showed that, compared with freezing survived S. aureus cells, the cells resuscitated for 1 h had 45 upregulated and 73 downregulated proteins. The differentially expressed proteins were functionally categorized by gene ontology enrichment, KEGG pathway, and STRING analyses. Cell membrane synthesis-related proteins, oxidative stress resistance-related proteins, metabolism-related proteins, and virulence factors exhibited distinct expression patterns during resuscitation. These findings have implications in the understanding of the resuscitation mechanism of freezing survived S. aureus, which may facilitate the development of novel technologies for improved detection and control of foodborne pathogens in frozen food. PMID:29774015

High-resolution scanning electron microscopy of frozen-hydrated cells.

PubMed

Walther, P; Chen, Y; Pech, L L; Pawley, J B

1992-11-01

Cryo-fixed yeast Paramecia and sea urchin embryos were investigated with an in-lens type field-emission SEM using a cold stage. The goal was to further develop and investigate the processing of frozen samples for the low-temperature scanning electron microscope (LTSEM). Uncoated frozen-hydrated samples were imaged with the low-voltage backscattered electron signal (BSE). Resolution and contrast were sufficient to visualize cross-fractured membranes, nuclear pores and small vesicles in the cytoplasm. It is assumed that the resolution of this approach is limited by the extraction depth of the BSE which depends upon the accelerating voltage of the primary beam (V0). In this study, the lowest possible V0 was 2.6 kV because below this value the sensitivity of the BSE detector is insufficient. It is concluded that the resolution of the uncoated specimen could be improved if equipment were available for high-resolution BSE imaging at 0.5-2 kV. Higher resolution was obtained with platinum cryo-coated samples, on which intramembranous particles were easily imaged. These images even show the ring-like appearance of the hexagonally arranged intramembranous particles known from high-resolution replica studies. On fully hydrated samples at high magnification, the observation time for a particular area is limited by mass loss caused by electron irradiation. Other potential sources of artefacts are the deposition of water vapour contamination and shrinkage caused by the sublimation of ice. Imaging of partially dehydrated (partially freeze-dried) samples, e.g. high-pressure frozen Paramecium and sea urchin embryos, will probably become the main application in cell biology. In spite of possible shrinkage problems, this approach has a number of advantages compared with any other electron microscopy preparation method: no chemical fixation is necessary, eliminating this source of artefacts; due to partial removal of the water additional structures in the cytoplasm can be investigated; and finally, the mass loss due to electron beam irradiation is greatly reduced compared to fully frozen-hydrated specimens.

Fabrication, characterization and fracture study of a machinable hydroxyapatite ceramic.

PubMed

Shareef, M Y; Messer, P F; van Noort, R

1993-01-01

In this study the preparation of a machinable hydroxyapatite from mixtures of a fine, submicrometer powder and either a coarse powder composed of porous aggregates up to 50 microns or a medium powder composed of dense particles of 3 microns median size is described. These were characterized using X-ray diffraction, transmission and scanning electron microscopy and infra-red spectroscopy. Test-pieces were formed by powder pressing and slip casting mixtures of various combinations of the fine, medium and coarse powders. The fired test-pieces were subjected to measurements of firing shrinkage, porosity, bulk density, tensile strength and fracture toughness. The microstructure and composition were examined using scanning electron microscopy and X-ray diffraction. For both processing methods, a uniform interconnected microporous structure was produced of a high-purity hydroxyapatite. The maximum tensile strength and fracture toughness that could be attained while retaining machinability were 37 MPa and 0.8 MPa m1/2 respectively.

Vacuum brazing of high volume fraction SiC particles reinforced aluminum matrix composites

NASA Astrophysics Data System (ADS)

Cheng, Dongfeng; Niu, Jitai; Gao, Zeng; Wang, Peng

2015-03-01

This experiment chooses A356 aluminum matrix composites containing 55% SiC particle reinforcing phase as the parent metal and Al-Si-Cu-Zn-Ni alloy metal as the filler metal. The brazing process is carried out in vacuum brazing furnace at the temperature of 550°C and 560°C for 3 min, respectively. The interfacial microstructures and fracture surfaces are investigated by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and energy spectrum analysis (EDS). The result shows that adequacy of element diffusion are superior when brazing at 560°C, because of higher activity and liquidity. Dislocations and twins are observed at the interface between filler and composite due to the different expansion coefficient of the aluminum alloy matrix and SiC particles. The fracture analysis shows that the brittle fracture mainly located at interface of filler and composites.

NCI Scientists Get Deep Look at CRISPR Complex Through Deep Freeze | Poster

Cancer.gov

To get a closer look at one CRISPR complex, researchers from NCI’s Center for Cancer Research and their collaborators recently put it “on ice” with cryo-electron microscopy, creating highly detailed images that show its biological structures in multiple states at a molecular level.

Cryopreservation of boar semen in mini- and maxi-straws.

PubMed

Bwanga, C O; de Braganca, M M; Einarsson, S; Rodriguez-Martinez, H

1990-10-01

Split ejaculates from four boars were frozen with a programmable freezing machine, in mini- (0.25 ml) and maxi- (5 ml) plastic straws with an extender at either acidic (6.3) or alkaline (7.4) pH. Glycerol (3%) was used as cryoprotectant. The freezing of the semen was monitored by way of thermocouples placed in the straws. Post-thaw motility and acrosome integrity were evaluated; the latter using phase contrast microscopy, eosin-nigrosin stain and electron microscopy. Post-thaw sperm motility was significantly higher when semen was frozen in mini-straws than in maxi-straws. For the mini-straws, the motility was better when semen was exposed to an acidic environment during freezing, but this beneficial effect of the low extracellular pH was not evident when maxi-straws were thawed. The motility of the spermatozoa diminished significantly during the thermoresistance test (0 h and 2 h time) at 37 degrees C in a similar way for both straws and extracellular pH's. The freezing procedure, no matter the extracellular pH, did not cause major acrosomal damages, but significantly more normal apical ridges were present in the mini-straws than in the maxi-straws. This in vitro evaluation indicated that the freezing method employed was better for mini- than for maxi-straws since the freezing of the 5 ml volumes was not homogeneous, due to the large section area between the surface and the core of the straw.

Development of structure in natural silk spinning and poly(vinyl alcohol) hydrogel formation

NASA Astrophysics Data System (ADS)

Willcox, Patricia Jeanene

This research involves the characterization of structure and structure formation in aqueous systems. Particularly, these studies investigate the effect of various processing variables on the structure formation that occurs upon conversion from aqueous solution to fiber or hydrogel. The two processes studied include natural silk fiber spinning and physical gelation of poly(vinyl alcohol), PVOH, in water. The techniques employed combine cryogenic technology for sample preparation and direct observation by transmission electron microscopy with electron diffraction, atomic force microscopy, optical rheometry, X-ray scattering and optical microscopy. In order to explore the full range of structure formation in natural silk spinning, studies are conducted in vivo and in vitro. In vivo structural investigations are accomplished through the cryogenic quenching and subsequent microtoming of live silk-spinning animals, Nephila clavipes (spider) and Bombyx mori (silkworm). Observations made using transmission electron microscopy, electron diffraction and atomic force microscopy indicate a cholesteric liquid crystalline mesophase of aqueous silk fibroin in both species. The mechanism of structure formation in solution is studied in vitro using optical rheometry on aqueous solutions made from regenerated Bombyx mori cocoon silk. Concentrated solutions exhibit birefringence under flow, with a wormlike conformation of the silk molecules in concentrated salt solution. Changes in salt concentration and pH of the aqueous silk solutions result in differing degrees of alignment and aggregation. These results suggest that structural control in the natural silk spinning process is accomplished by chemical manipulation of the electrostatic interactions and hydrogen bonding between chains. Application of cryogenic methods in transmission electron microscopy also provides a unique look at hydration-dependent structures in gels of poly(vinyl alcohol) produced by freeze-thaw processing. Morphologies ranging from circular pores to fibrillar networks are observed in gels formed from aqueous PVOH solutions subjected to cycles of freezing and thawing. These morphologies can be directly associated with the progressive nature of the mechanism of gelation as it proceeds from liquid-liquid phase separation to crystallization with increased cycling. A comparison of the structures produced by cycling and by aging suggests that there is a similarity in structural changes, but a superposition of the effects of cycling and aging is not possible.

Heave, settlement and fracture of chalk during physical modelling experiments with temperature cycling above and below 0 °C

NASA Astrophysics Data System (ADS)

Murton, Julian B.; Ozouf, Jean-Claude; Peterson, Rorik

2016-10-01

To elucidate the early stages of heave, settlement and fracture of intact frost-susceptible rock by temperature cycling above and below 0 °C, two physical modelling experiments were performed on 10 rectangular blocks 450 mm high of fine-grained, soft limestone. One experiment simulated 21 cycles of bidirectional freezing (upward and downward) of an active layer above permafrost, and the other simulated 26 cycles of unidirectional freezing (downward) of a seasonally frozen bedrock in a non-permafrost region. Heave and settlement of the top of the blocks were monitored in relation to rock temperature and unfrozen water content, which ranged from almost dry to almost saturated. In the bidirectional freezing experiment, heave of the wettest block initially occurred abruptly at the onset of freezing periods and gradually during thawing periods (summer heave). After the crossing of a threshold marked by the appearance of a macrocrack in the upper layer of permafrost, summer heave increased by an order of magnitude as segregated ice accumulated incrementally in macrocracks, interrupted episodically by abrupt settlement that coincided with unusually high air temperatures. In the unidirectional freezing experiment, the wet blocks heaved during freezing periods and settled during thawing periods, whereas the driest blocks showed the opposite behaviour. The two wettest blocks settled progressively during the first 15 freeze-thaw cycles, before starting to heave progressively as macrocracks developed. Four processes, operating singly or in combination in the blocks account for their heave and settlement: (1) thermal expansion and contraction caused heave and settlement when little or no water-ice phase change was involved; (2) volumetric expansion of water freezing in situ caused short bursts of heave of the outer millimetres of wet rock; (3) ice segregation deeper in the blocks caused sustained heave during thawing and freezing periods; and (4) freeze-thaw cycling caused consolidation and settlement of wet blocks prior to macrocracking in the unidirectional freezing experiment. Rock fracture developed by growth of segregated ice in microcracks and macrocracks at depths determined by the freezing regime. Overall, the heave, settlement and fracture behaviour of the limestone is similar to that of frost-susceptible soil.

Microstructure-Sensitive Investigation of Fracture Using Acoustic Emission Coupled With Electron Microscopy

NASA Technical Reports Server (NTRS)

Wisner, Brian; Cabal, Mike; Vanniamparambiland, Prashanth A.; Leser, William; Hochhalter, Jacob; Kontsos, Antonios

2015-01-01

A novel technique using Scanning Electron Microscopy (SEM) in conjunction with Acoustic Emission (AE) monitoring is proposed to investigate microstructure-sensitive fatigue and fracture of metals. The coupling between quasi in situ microscopy with actual in situ nondestructive evaluation falls into the ICME framework and the idea of quantitative data-driven characterization of material behavior. To validate the use of AE monitoring inside the SEM chamber, Aluminum 2024-B sharp notch specimen were tested both inside and outside the microscope using a small scale mechanical testing device. Subsequently, the same type of specimen was tested inside the SEM chamber. Load data were correlated with both AE information and observations of microcracks around grain boundaries as well as secondary cracks, voids, and slip bands. The preliminary results are in excellent agreement with similar findings at the mesoscale. Extensions of the application of this novel technique are discussed.

Robustness testing in pharmaceutical freeze-drying: inter-relation of process conditions and product quality attributes studied for a vaccine formulation.

PubMed

Schneid, Stefan C; Stärtzel, Peter M; Lettner, Patrick; Gieseler, Henning

2011-01-01

The recent US Food and Drug Administration (FDA) legislation has introduced the evaluation of the Design Space of critical process parameters in manufacturing processes. In freeze-drying, a "formulation" is expected to be robust when minor deviations of the product temperature do not negatively affect the final product quality attributes. To evaluate "formulation" robustness by investigating the effect of elevated product temperature on product quality using a bacterial vaccine solution. The vaccine solution was characterized by freeze-dry microscopy to determine the critical formulation temperature. A conservative cycle was developed using the SMART™ mode of a Lyostar II freeze dryer. Product temperature was elevated to imitate intermediate and aggressive cycle conditions. The final product was analyzed using X-ray powder diffraction (XRPD), scanning electron microscopy (SEM), Karl Fischer, and modulated differential scanning calorimetry (MDSC), and the life cell count (LCC) during accelerated stability testing. The cakes processed at intermediate and aggressive conditions displayed larger pores with microcollapse of walls and stronger loss in LCC than the conservatively processed product, especially during stability testing. For all process conditions, a loss of the majority of cells was observed during storage. For freeze-drying of life bacterial vaccine solutions, the product temperature profile during primary drying appeared to be inter-related to product quality attributes.

Cryo-Scanning Electron Microscopy (SEM) and Scanning Transmission Electron Microscopy (STEM)-in-SEM for Bio- and Organo-Mineral Interface Characterization in the Environment.

PubMed

Wille, Guillaume; Hellal, Jennifer; Ollivier, Patrick; Richard, Annie; Burel, Agnes; Jolly, Louis; Crampon, Marc; Michel, Caroline

2017-12-01

Understanding biofilm interactions with surrounding substratum and pollutants/particles can benefit from the application of existing microscopy tools. Using the example of biofilm interactions with zero-valent iron nanoparticles (nZVI), this study aims to apply various approaches in biofilm preparation and labeling for fluorescent or electron microscopy and energy dispersive X-ray spectrometry (EDS) microanalysis for accurate observations. According to the targeted microscopy method, biofilms were sampled as flocs or attached biofilm, submitted to labeling using 4',6-diamidino-2-phenylindol, lectins PNA and ConA coupled to fluorescent dye or gold nanoparticles, and prepared for observation (fixation, cross-section, freezing, ultramicrotomy). Fluorescent microscopy revealed that nZVI were embedded in the biofilm structure as aggregates but the resolution was insufficient to observe individual nZVI. Cryo-scanning electron microscopy (SEM) observations showed nZVI aggregates close to bacteria, but it was not possible to confirm direct interactions between nZVI and cell membranes. Scanning transmission electron microscopy in the SEM (STEM-in-SEM) showed that nZVI aggregates could enter the biofilm to a depth of 7-11 µm. Bacteria were surrounded by a ring of extracellular polymeric substances (EPS) preventing direct nZVI/membrane interactions. STEM/EDS mapping revealed a co-localization of nZVI aggregates with lectins suggesting a potential role of EPS in nZVI embedding. Thus, the combination of divergent microscopy approaches is a good approach to better understand and characterize biofilm/metal interactions.

Novel features of Brassica napus embryogenic microspores revealed by high pressure freezing and freeze substitution: evidence for massive autophagy and excretion-based cytoplasmic cleaning.

PubMed

Corral-Martínez, Patricia; Parra-Vega, Verónica; Seguí-Simarro, Jose M

2013-07-01

Induction of embryogenesis from isolated microspore cultures is a complex experimental system where microspores undergo dramatic changes in developmental fate. After ~40 years of application of electron microscopy to the study of the ultrastructural changes undergone by the induced microspore, there is still room for new discoveries. In this work, high pressure freezing and freeze substitution (HPF/FS), the best procedures known to date for ultrastructural preservation, were used to process Brassica napus microspore cultures covering all the stages of microspore embryogenesis. Analysis of these cultures by electron microscopy revealed massive processes of autophagy exclusively in embryogenic microspores, but not in other microspore-derived structures also present in cultures. However, a significant part of the autophagosomal cargo was not recycled. Instead, it was transported out of the cell, producing numerous deposits of extracytoplasmic fibrillar and membranous material. It was shown that commitment of microspores to embryogenesis is associated with both massive autophagy and excretion of the removed material. It is hypothesized that autophagy would be related to the need for a profound cytoplasmic cleaning, and excretion would be a mechanism to avoid excessive growth of the vacuolar system. Together, the results also demonstrate that the application of HPF/FS to the study of the androgenic switch is the best option currently available to identify the complex and dramatic ultrastructural changes undergone by the induced microspore. In addition, they provide significant insights to understand the cellular basis of induction of microspore embryogenesis, and open a new door for the investigation of this intriguing developmental pathway.

Intracellular Freezing in the Infective Juveniles of Steinernema feltiae: An Entomopathogenic Nematode

PubMed Central

Ali, Farman; Wharton, David A.

2014-01-01

Taking advantage of their optical transparency, we clearly observed the third stage infective juveniles (IJs) of Steinernema feltiae freezing under a cryo-stage microscope. The IJs froze when the water surrounding them froze at −2°C and below. However, they avoid inoculative freezing at −1°C, suggesting cryoprotective dehydration. Freezing was evident as a sudden darkening and cessation of IJs' movement. Freeze substitution and transmission electron microscopy confirmed that the IJs of S. feltiae freeze intracellularly. Ice crystals were found in every compartment of the body. IJs frozen at high sub-zero temperatures (−1 and −3°C) survived and had small ice crystals. Those frozen at −10°C had large ice crystals and did not survive. However, the pattern of ice formation was not well-controlled and individual nematodes frozen at −3°C had both small and large ice crystals. IJs frozen by plunging directly into liquid nitrogen had small ice crystals, but did not survive. This study thus presents the evidence that S. feltiae is only the second freeze tolerant animal, after the Antarctic nematode Panagrolaimus davidi, shown to withstand extensive intracellular freezing. PMID:24769523

Some personal and historical notes on the utility of “deep-etch” electron microscopy for making cell structure/function correlations

PubMed Central

Heuser, John E.

2014-01-01

This brief essay talks up the advantages of metal replicas for electron microscopy and explains why they are still the best way to image frozen cells in the electron microscope. Then it explains our approach to freezing, namely the Van Harreveld trick of “slamming” living cells onto a supercold block of metal sprayed with liquid helium at −269ºC, and further talks up this slamming over the alternative of high-pressure freezing, which is much trickier but enjoys greater favor at the moment. This leads me to bemoan the fact that there are not more young investigators today who want to get their hands on electron microscopes and use our approach to get the most “true to life” views of cells out of them with a minimum of hassle. Finally, it ends with a few perspectives on my own career and concludes that, personally, I'm permanently stuck with the view of the “founding fathers” that cell ultrastructure will ultimately display and explain all of cell function, or as Palade said in his Nobel lecture,electron micrographs are “irresistible and half transparent … their meaning buried under only a few years of work,” and “reasonable working hypotheses are already suggested by the ultrastructural organization itself.” PMID:25360049

Fracture modes in notched angleplied composite laminates

NASA Technical Reports Server (NTRS)

Irvine, T. B.; Ginty, C. A.

1984-01-01

The Composite Durability Structural Analysis (CODSTRAN) computer code is used to determine composite fracture. Fracture modes in solid and notched, unidirectional and angleplied graphite/epoxy composites were determined by using CODSTRAN. Experimental verification included both nondestructive (ultrasonic C-Scanning) and destructive (scanning electron microscopy) techniques. The fracture modes were found to be a function of ply orientations and whether the composite is notched or unnotched. Delaminations caused by stress concentrations around notch tips were also determined. Results indicate that the composite mechanics, structural analysis, laminate analysis, and fracture criteria modules embedded in CODSTRAN are valid for determining composite fracture modes.

Scanning electron microscopy combined with image processing technique: Analysis of microstructure, texture and tenderness in Semitendinous and Gluteus Medius bovine muscles.

PubMed

Pieniazek, Facundo; Messina, Valeria

2016-11-01

In this study the effect of freeze drying on the microstructure, texture, and tenderness of Semitendinous and Gluteus Medius bovine muscles were analyzed applying Scanning Electron Microscopy combined with image analysis. Samples were analyzed by Scanning Electron Microscopy at different magnifications (250, 500, and 1,000×). Texture parameters were analyzed by Texture analyzer and by image analysis. Tenderness by Warner-Bratzler shear force. Significant differences (p < 0.05) were obtained for image and instrumental texture features. A linear trend with a linear correlation was applied for instrumental and image features. Image texture features calculated from Gray Level Co-occurrence Matrix (homogeneity, contrast, entropy, correlation and energy) at 1,000× in both muscles had high correlations with instrumental features (chewiness, hardness, cohesiveness, and springiness). Tenderness showed a positive correlation in both muscles with image features (energy and homogeneity). Combing Scanning Electron Microscopy with image analysis can be a useful tool to analyze quality parameters in meat.Summary SCANNING 38:727-734, 2016. © 2016 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

Experiments in electron microscopy: from metals to nerves

NASA Astrophysics Data System (ADS)

Unwin, Nigel

2015-04-01

Electron microscopy has advanced remarkably as a tool for biological structure research since the development of methods to examine radiation-sensitive unstained specimens and the introduction of cryo-techniques. Structures of biological molecules at near-atomic resolution can now be obtained from images of single particles as well as crystalline arrays. It has also become possible to analyze structures of molecules in their functional context, i.e. in their natural membrane or cellular setting, and in an ionic environment like that in living tissue. Electron microscopy is thus opening ways to answer definitively questions about physiological mechanisms. Here I recall a number of experiments contributing to, and benefiting from the technical advances that have taken place. I begin—in the spirit of this crystallography series—with some biographical background, and then sketch the path to an analysis by time-resolved microscopy of the opening mechanism of an ion channel (nicotinic acetylcholine receptor). This analysis illustrates how electron imaging can be combined with freeze-trapping to illuminate a transient biological event: in our case, chemical-to-electrical transduction at the nerve-muscle synapse.

Bending fracture in carbon nanotubes.

PubMed

Kuo, Wen-Shyong; Lu, Hsin-Fang

2008-12-10

A novel approach was adopted to incur bending fracture in carbon nanotubes (CNTs). Expanded graphite (EG) was made by intercalating and exfoliating natural graphite flakes. The EG was deposited with nickel particles, from which CNTs were grown by chemical vapor deposition. The CNTs were tip-grown, and their roots were fixed on the EG flakes. The EG flakes were compressed, and many CNTs on the surface were fragmented due to the compression-induced bending. Two major modes of the bending fracture were observed: cone-shaped and shear-cut. High-resolution scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to examine the crack growth within the graphene layers. The bending fracture is characterized by two-region crack growth. An opening crack first appears around the outer-tube due to the bending-induced tensile stress. The crack then branches to grow along an inclined direction toward the inner-tube due to the presence of the shear stress in between graphene layers. An inner-tube pullout with inclined side surface is formed. The onset and development of the crack in these two regions are discussed.

Effects of Core-Shell Rubber (CSR) Nanoparticles on the Cryogenic Fracture Toughness of CSR Modified Epoxy

NASA Technical Reports Server (NTRS)

Wang, Jun; Magee, Daniel; Schneider, Judy; Cannon, Seth

2009-01-01

This study investigated the effects of core-shell rubber (CSR) nanoparticles on the mechanical properties and fracture toughness of an epoxy resin at ambient and liquid nitrogen (LN2) temperatures. Varying amounts of Kane Ace(Registered TradeMark) MX130 and Kane Ace(Registered TradeMark) MX960 toughening agent were added to a commercially available EPON 862/Epikure W epoxy resin. Elastic modulus was calculated using quasi-static tensile data. Fracture toughness was evaluated by the resulting breaking energy measured in Charpy impact tests conducted on an instrumented drop tower. The size and distribution of the CSR nanoparticles were characterized using Transmission Electron Microscopy (TEM) and Small Angle X-ray Scattering (SAXS). Scanning Electron Microscopy (SEM) was used to study the fracture surface morphology. The addition of the CSR nanoparticles increased the breaking energy with negligible change in elastic modulus and ultimate tensile stress (UTS). At ambient temperature the breaking energy increased with increasing additions of the CSR nanoparticles up to 13.8wt%, while at LN2 temperatures, it reached a plateau at much lower CSR concentration.

Effects of Pressure-shift Freezing on the Structural and Physical Properties of Gelatin Hydrogel Matrices

PubMed Central

Kim, Byeongsoo; Gil, Hyung Bae; Min, Sang-Gi; Lee, Si-Kyung; Choi, Mi-Jung

2014-01-01

This study investigates the effects of the gelatin concentration (10-40%, w/v), freezing temperatures (from -20℃ to -50℃) and freezing methods on the structural and physical properties of gelatin matrices. To freeze gelatin, the pressure-shift freezing (PSF) is being applied at 0.1 (under atmospheric control), 50 and 100 MPa, respectively. The freezing point of gelatin solutions decrease with increasing gelatin concentrations, from -0.2℃ (10% gelatin) to -6.7℃ (40% gelatin), while the extent of supercooling did not show any specific trends. The rheological properties of the gelatin indicate that both the storage (G') and loss (G") moduli were steady in the strain amplitude range of 0.1-10%. To characterize gelatin matrices formed by the various freezing methods, the ice crystal sizes which were being determined by the scanning electron microscopy (SEM) are affected by the gelatin concentrations. The ice crystal sizes are affected by gelatin concentrations and freezing temperature, while the size distributions of ice crystals depend on the freezing methods. Smaller ice crystals are being formed with PSF rather than under the atmospheric control where the freezing temperature is above -40℃. Thus, the results of this study indicate that the PSF processing at a very low freezing temperature (-50℃) offers a potential advantage over commercial atmospheric freezing points for the formation of small ice crystals. PMID:26760743

Establishment of a Human Blood-Brain Barrier Co-culture Model Mimicking the Neurovascular Unit Using Induced Pluri- and Multipotent Stem Cells.

PubMed

Appelt-Menzel, Antje; Cubukova, Alevtina; Günther, Katharina; Edenhofer, Frank; Piontek, Jörg; Krause, Gerd; Stüber, Tanja; Walles, Heike; Neuhaus, Winfried; Metzger, Marco

2017-04-11

In vitro models of the human blood-brain barrier (BBB) are highly desirable for drug development. This study aims to analyze a set of ten different BBB culture models based on primary cells, human induced pluripotent stem cells (hiPSCs), and multipotent fetal neural stem cells (fNSCs). We systematically investigated the impact of astrocytes, pericytes, and NSCs on hiPSC-derived BBB endothelial cell function and gene expression. The quadruple culture models, based on these four cell types, achieved BBB characteristics including transendothelial electrical resistance (TEER) up to 2,500 Ω cm 2 and distinct upregulation of typical BBB genes. A complex in vivo-like tight junction (TJ) network was detected by freeze-fracture and transmission electron microscopy. Treatment with claudin-specific TJ modulators caused TEER decrease, confirming the relevant role of claudin subtypes for paracellular tightness. Drug permeability tests with reference substances were performed and confirmed the suitability of the models for drug transport studies. Copyright © 2017 The Author(s). Published by Elsevier Inc. All rights reserved.

Formation of cage-like particles by poly(amino acid)-based block copolymers in aqueous solution.

PubMed Central

Cudd, A; Bhogal, M; O'Mullane, J; Goddard, P

1991-01-01

When dissolved in N,N-dimethylformamide and then dialyzed against phosphate-buffered saline, A-B-A block copolymers composed of poly [N5-(2-hydroxyethyl)-L-glutamine]-block-poly(gamma-benzyl-L-glutamate)- block-poly [N5-(2-hydroxyethyl)-L-glutamine] form particles. The particles are cage-like structures with average diameters of 300 nm (average polydispersity, 0.3-0.5). They are stable in aqueous solution at 4 degrees C for up to 3 weeks, at which time flocculation becomes apparent. Negative staining and freeze-fracture electron microscopy suggest that cage-like particles are formed by selective association of segregated micelle populations. A model of particle formation is presented in which B blocks form micelles in dimethylformamide. On dialysis against an aqueous solution, the extended A blocks then associate intermolecularly to form rod-shaped micelles, which connect the B block micelles. The result is a meshed cage-like particle. The implications of these observations on the aggregation behavior of polymeric surfactants in dilute solution are discussed. Images PMID:11607245

Supramolecular "Big Bang" in a Single-Ionic Surfactant/Water System Driven by Electrostatic Repulsion: From Vesicles to Micelles.

PubMed

Leclercq, Loïc; Bauduin, Pierre; Nardello-Rataj, Véronique

2017-04-11

In aqueous solution, dimethyldi-n-octylammonium chloride, [DiC 8 ][Cl], spontaneously forms dimers at low concentrations (1-10 mM) to decrease the strength of the hydrophobic-water contact. Dimers represent ideal building blocks for the abrupt edification of vesicles at 10 mM. These vesicles are fully characterized by dynamic and static light scattering, self-diffusion nuclear magnetic resonance, and freeze-fracture transmission electron microscopy. An increase in concentration leads to electrostatic repulsion between vesicles that explode into small micelles at 30 mM. These transitions are detected by means of surface tension, conductivity, and solubility of hydrophobic solutes as well as by isothermal titration microcalorimetry. These unusual supramolecular transitions emerge from the surfactant chemical structure that combines two contradictory features: (i) the double-chain structure tending to form low planar aggregates with low water solubility and (ii) the relatively short chains giving high hydrophilicity. The well-balanced hydrophilic-hydrophobic character of [DiC 8 ][Cl] is then believed to be at the origin of the unusual supramolecular sequence offering new opportunities for drug delivery systems.

Immunoelectron Microscopy of Cryofixed and Freeze-Substituted Plant Tissues.

PubMed

Takeuchi, Miyuki; Takabe, Keiji; Mineyuki, Yoshinobu

2016-01-01

Cryofixation and freeze-substitution techniques provide excellent preservation of plant ultrastructure. The advantage of cryofixation is not only in structural preservation, as seen in the smooth plasma membrane, but also in the speed in arresting cell activity. Immunoelectron microscopy reveals the subcellular localization of molecules within cells. Immunolabeling in combination with cryofixation and freeze-substitution techniques provides more detailed information on the immunoelectron-microscopic localization of molecules in the plant cell than can be obtained from chemically fixed tissues. Here, we introduce methods for immunoelectron microscopy of cryofixed and freeze-substituted plant tissues.

Bridging Philosophy of Technology and Neurobiological Research: Interpreting Images from the "Slam Freezer"

ERIC Educational Resources Information Center

Rosenberger, Robert

2005-01-01

The swiftly growing field of neurobiological research utilizes highly advanced technologies (e.g., magnetic resonance imaging, electron microscopy) to mediate between investigators and the brains they investigate. Here, the author analyzes a device called the "slam freezer" that quick-freezes neurons to be studied under the microscope. Employing…

Cold-stage microscopy system for fast-frozen liquids.

PubMed

Talmon, Y; Davis, H T; Scriven, L E; Thomas, E L

1979-06-01

The least artifact-laden fixation technique for examining colloidal suspensions, microemulsions, and other microstructured liquids in the electron microscope appears to be thermal fixation, i.e., ultrafast freezing of the liquid specimen. For rapid-enough cooling and for observation in TEM/STEM a thin sample is needed. The need is met by trapping a thin layer ( approximately 100 nm) of liquid between two polyimide films ( approximately 40 nm thickness) mounted on copper grids and immersing the resulting sandwich in liquid nitrogen at its melting point. For liquids containing water, polyimides films are used since this polymer is far less susceptible to the electron beam damage observed for the commonly used polymer films such as Formvar and collodion in contact with ice. Transfer of the frozen sample into the microscope column without deleterious frost deposition and warming is accomplished with a new transfer module for the cooling stage of the JEOL JEM-100CX microscope, which makes a true cold stage out of a device originally intended for cooling specimens inside the column. Sample results obtained with the new fast-freeze, cold-stage microscopy system are given.

Development of formulations and processes to incorporate wax oleogels in ice cream.

PubMed

Zulim Botega, Daniele C; Marangoni, Alejandro G; Smith, Alexandra K; Goff, H Douglas

2013-12-01

The objective of this study was to investigate the influence of emulsifiers, waxes, fat concentration, and processing conditions on the application of wax oleogel to replace solid fat content and create optimal fat structure in ice cream. Ice creams with 10% or 15% fat were formulated with rice bran wax (RBW), candelilla wax (CDW), or carnauba wax (CBW) oleogels, containing 10% wax and 90% high-oleic sunflower oil. The ice creams were produced using batch or continuous freezing processes. Transmission electron microscopy (TEM) and cryo-scanning electron microscopy were used to evaluate the microstructure of ice cream and the ultrastructure of oleogel droplets in ice cream mixes. Among the wax oleogels, RBW oleogel had the ability to form and sustain structure in 15% fat ice creams when glycerol monooleate (GMO) was used as the emulsifier. TEM images revealed that the high degree of fat structuring observed in GMO samples was associated with the RBW crystal morphology within the fat droplet, which was characterized by the growth of crystals at the outer edge of the droplet. Continuous freezing improved fat structuring compared to batch freezing. RBW oleogels established better structure compared to CDW or CBW oleogels. These results demonstrate that RBW oleogel has the potential to develop fat structure in ice cream in the presence of GMO and sufficiently high concentrations of oleogel. © 2013 Institute of Food Technologists®

Fractography of induction-hardened steel fractured in fatigue and overload

DOE Office of Scientific and Technical Information (OSTI.GOV)

Santos, C.G.; Laird, C.

1997-07-01

The fracture surfaces of induction-hardened steel specimens obtained from an auto axle were characterized, macroscopically and microscopically, after being fractured in fatigue and monotonic overload. Specimens were tested in cyclic three-point bending under load control, and the S-N curve was established for specimens that had been notched by spark machining to facilitate fractography. Scanning electron microscopy of the fractured surfaces obtained for lives spanning the range 17,000 to 418,000 cycles revealed diverse fracture morphologies, including intergranular fracture and transgranular fatigue fracture. The results are being offered to assist in the analysis of complex field failures in strongly hardened steel.

Development of the gametophyte of the fern Schizaea pusilla

NASA Technical Reports Server (NTRS)

Kiss, J. Z.; Swatzell, L. J.

1996-01-01

Schizaea pusilla is a pteridophyte with several unique developmental characteristics. In contrast to most other fern species, S. pusilla gametophytes remain filamentous throughout their development, and the gametophytes are associated with an endophytic fungus which appears to be mycorrhizal. In terms of tropistic responses, apical filament cells of young gametophytes are negatively phototropic compared with germ filaments of other ferns which exhibit positive phototropism. Cryofixation (propane jet freezing and high-pressure freezing) in conjunction with freeze substitution electron microscopy was used to study young gametophytes. The results demonstrate that apical filament cells have a distinctive structural polarity and that rhizoids also can be successfully frozen by these methods. The cytoskeleton and endomembrane system were particularly well preserved in cryofixed cells. In addition, Schizaea gametophytes were used as a test system to evaluate potential artifacts of propane jet freezing and high pressure freezing. There was little apparent difference in ultrastructure between cells cryofixed by either freezing method. These gametophytes will be useful in determining the effectiveness of cryofixation techniques and as a model system in tip growth studies.

Inner ear tissue preservation by rapid freezing: Improving fixation by high-pressure freezing and hybrid methods

PubMed Central

Bullen, A.; Taylor, R.R.; Kachar, B.; Moores, C.; Fleck, R.A.; Forge, A.

2014-01-01

In the preservation of tissues in as ‘close to life’ state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40 μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues. PMID:25016142

Application of freeze-drying technology in manufacturing orally disintegrating films.

PubMed

Liew, Kai Bin; Odeniyi, Michael Ayodele; Peh, Kok-Khiang

2016-01-01

Freeze drying technology has not been maximized and reported in manufacturing orally disintegrating films. The aim of this study was to explore the freeze drying technology in the formulation of sildenafil orally disintegrating films and compare the physical properties with heat-dried orally disintegrating film. Central composite design was used to investigate the effects of three factors, namely concentration of carbopol, wheat starch and polyethylene glycol 400 on the tensile strength and disintegration time of the film. Heat-dried films had higher tensile strength than films prepared using freeze-dried method. For folding endurance, freeze-dried films showed improved endurance than heat-dried films. Moreover, films prepared using freeze-dried methods were thicker and had faster disintegration time. Formulations with higher amount of carbopol and starch showed higher tensile strength and thickness whereas formulations with higher PEG 400 content showed better flexibility. Scanning electron microscopy showed that the freeze-dried films had more porous structure compared to the heat-dried film as a result of the release of water molecule from the frozen structure when it was subjected to freeze drying process. The sildenafil film was palatable. The dissolution profiles of freeze-dried and heat-dried films were similar to Viagra® with f2 of 51.04 and 65.98, respectively.

Overexpression of Arabidopsis NADPH-dependent thioredoxin reductase C (AtNTRC) confers freezing and cold shock tolerance to plants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Moon, Jeong Chan; Lee, Sangmin; Shin, Su Young

Overexpression of AtNTRC (AtNTRC{sup OE}) in Arabidopsis thaliana led to a freezing and cold stress tolerance, whereas a knockout mutant (atntrc) showed a stress-sensitive phenotype. Biochemical analyses showed that the recombinant AtNTRC proteins exhibited a cryoprotective activity for malate dehydrogenase and lactic dehydrogenase. Furthermore, conclusive evidence of its interaction with nucleic acids in vitro is provided here on the basis of gel shift and electron microscopy analysis. Recombinant AtNTRC efficiently protected RNA and DNA from RNase A and metal catalyzed oxidation damage, respectively. The C-terminal thioredoxin domain is required for the nucleic acid–protein complex formation. From these results, it can bemore » hypothesized that AtNTRC, which is known to be an electron donor of peroxiredoxin, contributes the stability of macromolecules under cold stress. - Highlights: • AtNTRC has a cryoprotective activity in vitro. • Overexpression of AtNTRC increases tolerance to freezing and cold shock stresses. • Thioredoxin domain of AtNTRC protects nucleic acids in vitro. • AtNTRC inhibits protein aggregation under freezing stress in vitro.« less

Dynamic fracture toughness of cellulose-fiber-reinforced polypropylene : preliminary investigation of microstructural effects

Treesearch

Craig M. Clemons; Daniel F. Caulfield; A. Jeffrey Giacomin

1999-10-01

In this study, the microstructure of injection-molded polypropylene reinforced with cellulose fiber was investigated. Scanning electron microscopy of the fracture surfaces and X-ray diffraction were used to investigate fiber orientation. The polypropylene matrix was removed by solvent extraction, and the lengths of the residual fibers were optically determined. Fiber...

Making and breaking bridges in a Pickering emulsion.

PubMed

French, David J; Taylor, Phil; Fowler, Jeff; Clegg, Paul S

2015-03-01

Particle bridges form in Pickering emulsions when the oil-water interfacial area generated by an applied shear is greater than that which can be stabilised by the available particles and the particles have a slight preference for the continuous phase. They can subsequently be broken by low shear or by modifying the particle wettability. We have developed a model oil-in-water system for studying particle bridging in Pickering emulsions stabilised by fluorescent Stöber silica. A mixture of dodecane and isopropyl myristate was used as the oil phase. We have used light scattering and microscopy to study the degree to which emulsions are bridged, and how this is affected by parameters including particle volume fraction, particle wettability and shear rate. We have looked for direct evidence of droplets sharing particles using freeze fracture scanning electron microscopy. We have created strongly aggregating Pickering emulsions using our model system. This aggregating state can be accessed by varying several different parameters, including particle wettability and particle volume fraction. Particles with a slight preference for the continuous phase are required for bridging to occur, and the degree of bridging increases with increasing shear rate but decreases with increasing particle volume fraction. Particle bridges can subsequently be removed by applying low shear or by modifying the particle wettability. Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.

Application of scanning acoustic microscopy to advanced structural ceramics

NASA Technical Reports Server (NTRS)

Vary, Alex; Klima, Stanley J.

1987-01-01

A review is presentod of research investigations of several acoustic microscopy techniques for application to structural ceramics for advanced heat engines. Results obtained with scanning acoustic microscopy (SAM), scanning laser acoustic microscopy (SLAM), scanning electron acoustic microscopy (SEAM), and photoacoustic microscopy (PAM) are compared. The techniques were evaluated on research samples of green and sintered monolithic silicon nitrides and silicon carbides in the form of modulus-of-rupture bars containing deliberately introduced flaws. Strengths and limitations of the techniques are described with emphasis on statistics of detectability of flaws that constitute potential fracture origins.

Accurate prediction of collapse temperature using optical coherence tomography-based freeze-drying microscopy.

PubMed

Greco, Kristyn; Mujat, Mircea; Galbally-Kinney, Kristin L; Hammer, Daniel X; Ferguson, R Daniel; Iftimia, Nicusor; Mulhall, Phillip; Sharma, Puneet; Kessler, William J; Pikal, Michael J

2013-06-01

The objective of this study was to assess the feasibility of developing and applying a laboratory tool that can provide three-dimensional product structural information during freeze-drying and which can accurately characterize the collapse temperature (Tc ) of pharmaceutical formulations designed for freeze-drying. A single-vial freeze dryer coupled with optical coherence tomography freeze-drying microscopy (OCT-FDM) was developed to investigate the structure and Tc of formulations in pharmaceutically relevant products containers (i.e., freeze-drying in vials). OCT-FDM was used to measure the Tc and eutectic melt of three formulations in freeze-drying vials. The Tc as measured by OCT-FDM was found to be predictive of freeze-drying with a batch of vials in a conventional laboratory freeze dryer. The freeze-drying cycles developed using OCT-FDM data, as compared with traditional light transmission freeze-drying microscopy (LT-FDM), resulted in a significant reduction in primary drying time, which could result in a substantial reduction of manufacturing costs while maintaining product quality. OCT-FDM provides quantitative data to justify freeze-drying at temperatures higher than the Tc measured by LT-FDM and provides a reliable upper limit to setting a product temperature in primary drying. Copyright © 2013 Wiley Periodicals, Inc.

Characterization of Propylene Glycol-Mitigated Freeze/Thaw Agglomeration of a Frozen Liquid nOMV Vaccine Formulation by Static Light Scattering and Micro-Flow Imaging.

PubMed

Mensch, Christopher D; Davis, Harrison B; Blue, Jeffrey T

2015-01-01

The purpose of this work was to investigate the susceptibility of an aluminum adjuvant and an aluminum-adjuvanted native outer membrane vesicle (nOMV) vaccine formulation to freeze/thaw-induced agglomeration using static light scattering and micro-flow Imaging analysis; and to evaluate the use of propylene glycol as a vaccine formulation excipient by which freeze/thaw-induced agglomeration of a nOMV vaccine formulation could be mitigated. Our results indicate that including 7% v/v propylene glycol in an nOMV containing aluminum adjuvanted vaccine formulation, mitigates freeze/thaw-induced agglomeration. We evaluated the effect of freeze-thawing on an aluminum adjuvant and an aluminum adjuvanted native outer membrane vesicle (nOMV) vaccine formulation. Specifically, we characterized the freeze/thaw-induced agglomeration through the use of static light scattering, micro-flow imaging, and cryo-electron microscopy analysis. Further, we evaluated the use of 0-9% v/v propylene glycol as an excipient which could be included in the formulation for the purpose of mitigating the agglomeration induced by freeze/thaw. The results indicate that using 7% v/v propylene glycol as a formulation excipient is effective at mitigating agglomeration of the nOMV vaccine formulation, otherwise induced by freeze-thawing. © PDA, Inc. 2015.

Enhanced stress corrosion cracking resistance and electrical conductivity of a T761 treated Al-Zn-Mg-Cu alloy thin plate

NASA Astrophysics Data System (ADS)

Chen, Xu; Zhai, Sudan; Gao, Di; Liu, Ye; Xu, Jing; Liu, Yang

2018-01-01

The stress corrosion cracking (SCC) behavior, electrical conductivity and mechanical properties of an Al-Zn-Mg-Cu alloy pre-stretched thin plate for wing skin were researched in this paper. The microstructures and SCC fracture surfaces of the alloy treated at different conditions were characterized by transmission electron microscopy, optical microscopy and scanning electron microscopy. Results indicated that with the increasing of aging temperature, the electrical conductivity and the elongation increased greatly, while the strength decreased gradually which were closely associated with the type and morphology of the precipitates. Compared with the T6 treated alloy, the SCC resistance of the T761 treated Al-Zn-Mg-Cu alloy was improved greatly. The SCC behavior of the T6 treated alloy was dominated by anodic dissolution theory, whereas the hydrogen induced cracking controlled the fracture behavior of the T761 treated alloy which was influenced by the morphology of grain boundary precipitates in this investigated alloy.

Cellular control over spicule formation in sea urchin embryos: A structural approach.

PubMed

Beniash, E; Addadi, L; Weiner, S

1999-03-01

The spicules of the sea urchin embryo form in intracellular membrane-delineated compartments. Each spicule is composed of a single crystal of calcite and amorphous calcium carbonate. The latter transforms with time into calcite by overgrowth of the preexisting crystal. Relationships between the membrane surrounding the spiculogenic compartment and the spicule mineral phase were studied in the transmission electron microscope (TEM) using freeze-fracture. In all the replicas observed the spicules were tightly surrounded by the membrane. Furthermore, a variety of structures that are related to the material exchange process across the membrane were observed. The spiculogenic cells were separated from other cell types of the embryo, frozen, and freeze-dried on the TEM grids. The contents of electron-dense granules in the spiculogenic cells were shown by electron diffraction to be composed of amorphous calcium carbonate. These observations are consistent with the notion that the amorphous calcium carbonate-containing granules contain the precursor mineral phase for spicule formation and that the membrane surrounding the forming spicule is involved both in transport of material and in controlling spicule mineralization. Copyright 1999 Academic Press.

Size-dependent fracture mode transition in copper nanowires.

PubMed

Peng, Cheng; Zhan, Yongjie; Lou, Jun

2012-06-25

In situ uni-axial tensile tests of single-crystalline copper nanowires are performed using a micromechanical device inside a scanning electron microscope chamber. The single-crystalline copper nanowires are synthesized by solvothermal processes, and the growth direction along the wire axis is the <110> orientation as confirmed by transmission electron microscopy (TEM) selected area diffraction (SAD) analysis. The fracture strengths of copper nanowires are found to be much higher than that of bulk copper. More interestingly, both ductile and brittle-like fracture modes are found in the same batch of fabricated nanowires, and the fracture modes appear to be dependent on the diameters of tested nanowires. From the analysis of fracture surfaces, sample morphologies and corresponding stress-strain curves, the competition between deformation and fracture mechanisms controlled by initial defects density and by the probability of dislocation interactions is attributed to this intriguing size-dependent fracture mode transition. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Three-dimensional imaging of adherent cells using FIB/SEM and STEM.

PubMed

Villinger, Clarissa; Schauflinger, Martin; Gregorius, Heiko; Kranz, Christine; Höhn, Katharina; Nafeey, Soufi; Walther, Paul

2014-01-01

In this chapter we describe three different approaches for three-dimensional imaging of electron microscopic samples: serial sectioning transmission electron microscopy (TEM), scanning transmission electron microscopy (STEM) tomography, and focused ion beam/scanning electron microscopy (FIB/SEM) tomography. With these methods, relatively large volumes of resin-embedded biological structures can be analyzed at resolutions of a few nm within a reasonable expenditure of time. The traditional method is serial sectioning and imaging the same area in all sections. Another method is TEM tomography that involves tilting a section in the electron beam and then reconstruction of the volume by back projection of the images. When the scanning transmission (STEM) mode is used, thicker sections (up to 1 μm) can be analyzed. The third approach presented here is focused ion beam/scanning electron microscopy (FIB/SEM) tomography, in which a sample is repeatedly milled with a focused ion beam (FIB) and each newly produced block face is imaged with the scanning electron microscope (SEM). This process can be repeated ad libitum in arbitrary small increments allowing 3D analysis of relatively large volumes such as eukaryotic cells. We show that resolution of this approach is considerably improved when the secondary electron signal is used. However, the most important prerequisite for three-dimensional imaging is good specimen preparation. For all three imaging methods, cryo-fixed (high-pressure frozen) and freeze-substituted samples have been used.

A novel adhering junction in the apical ciliary apparatus of the rotifer Brachionus plicatilis (Rotifera, Monogononta).

PubMed

Dallai, R; Lupetti, P; Lane, N J

1996-10-01

Cultures of the rotifer Brachionus plicatilis were examined with regard to their interepithelial junctions after infiltration with the extracellular tracer lanthanum, freeze-fracturing or quick-freeze deep-etching. The lateral borders between ciliated cells have an unusual apical adhering junction. This apical part of their intercellular cleft looks desmosome-like, but it is characterized by unusual intramembranous E-face clusters of particles. Deep-etching reveals that these are packed together in short rows which lie parallel to one another in orderly arrays. The true membrane surface in these areas features filaments in the form of short ribbons; these are produced by projections, possibly part of the glycocalyx, emerging from the membranes, between which the electron-dense tracer lanthanum permeates. These projections appear to overlap with each other in the centre of the intercellular cleft; this would provide a particularly flexible adaptation to maintain cell-cell contact and coordination as a consequence. The filamentous ribbons may be held in position by the intramembranous particle arrays since both have a similar size and distribution. These contacts are quite different from desmosomes and appear to represent a distinct new category of adhesive cell-cell junction. Beneath these novel structures, conventional pleated septate junctions are found, exhibiting the undulating intercellular ribbons typical of this junctional type, as well as the usual parallel alignments of intramembranous rows of EF grooves and PF particles. Below these are found gap junctions as close-packed plaques of intramembranous particles on either the P-face or E-face. After freeze-fracturing, the complementary fracture face to the particles shows pits, usually on the P-face, arrayed with a very precise hexagonal pattern.

Novel Method to Characterize and Model the Multiaxial Constitutive and Damage Response of Energetic Materials.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kaneshige, Michael J.; Rabbi, Md Fazle; Kaneshige, Michael J.

2017-12-01

Simulant polymer bonded explosives are widely used to simulate the mechanical response of real energetic materials. In this paper, the fracture resistance of a simulant polymer bo nded explosive (PBX) is experimentally investigated. The simulant is composed of 80 wt.% soda lime glass beads (SLGB) and 20 wt.% high impact Polystyrene 825 (HIPS). Brazilian disk tests are performed to characterize the tensile and compressive properties. Fracture toughness and energy tests are performed in the semi - circular bending (SCB) configuration on 80, 81, 82, and 83 wt % SLGB compositions. Digital image correlation is performed to record the surface displacementsmore » and calculate surface strains during testing. The m icromechanical behavior of ductile and brittle fracture are evaluated using digital microscopy and scanning electron microscopy of the fracture surface. It is determined that (i) the manufacturing process produces a credible simulant of PBX properties, and (ii) the SCB test measures fracture resistance with a reasonable coefficient of variation.« less

Inner ear tissue preservation by rapid freezing: improving fixation by high-pressure freezing and hybrid methods.

PubMed

Bullen, A; Taylor, R R; Kachar, B; Moores, C; Fleck, R A; Forge, A

2014-09-01

In the preservation of tissues in as 'close to life' state as possible, rapid freeze fixation has many benefits over conventional chemical fixation. One technique by which rapid freeze-fixation can be achieved, high pressure freezing (HPF), has been shown to enable ice crystal artefact-free freezing and tissue preservation to greater depths (more than 40 μm) than other quick-freezing methods. Despite increasingly becoming routine in electron microscopy, the use of HPF for the fixation of inner ear tissue has been limited. Assessment of the quality of preservation showed routine HPF techniques were suitable for preparation of inner ear tissues in a variety of species. Good preservation throughout the depth of sensory epithelia was achievable. Comparison to chemically fixed tissue indicated that fresh frozen preparations exhibited overall superior structural preservation of cells. However, HPF fixation caused characteristic artefacts in stereocilia that suggested poor quality freezing of the actin bundles. The hybrid technique of pre-fixation and high pressure freezing was shown to produce cellular preservation throughout the tissue, similar to that seen in HPF alone. Pre-fixation HPF produced consistent high quality preservation of stereociliary actin bundles. Optimising the preparation of samples with minimal artefact formation allows analysis of the links between ultrastructure and function in inner ear tissues. Copyright © 2014 The Authors. Published by Elsevier B.V. All rights reserved.

Study of poly(N,N-dimethylacrylamide)/CdS nanocomposite organic/inorganic gels.

PubMed

Bekiari, Vlasoula; Pagonis, Konstantinos; Bokias, Georgios; Lianos, Panagiotis

2004-09-14

CdS nanoparticles have been synthesized and stabilized in poly(N,N-dimethylacrylamide) hydrogels. The properties of the composite material have been characterized by UV-vis spectroscopy, scanning electron microscopy, X-ray diffraction, thermogravimetric analysis, and steady-state and time-resolved luminescence spectroscopy. This material can be obtained in three different states: swollen, shrunk, and freeze-dried. The swollen and the freeze-dried states correspond to a nanocomposite organic/inorganic (wet or dry) gel containing CdS nanoparticles of approximately 50 nm diameter while the shrunk state is a two-phase system containing CdS crystals, which precipitate forming interesting geometrical shapes.

Investigation of the plastic fracture of high strength steels

NASA Technical Reports Server (NTRS)

Cox, T. B.; Low, J. R., Jr.

1972-01-01

This investigation deals in detail with the three recognized stages of plastic fracture in high strength steels, namely, void initiation, void growth, and void coalescence. The particular steels under investigation include plates from both commercial purity and high purity heats of AISI 4340 and 18 Ni, 200 grade maraging steels. A scanning electron microscope equipped with an X-ray energy dispersive analyzer, together with observations made using light microscopy, revealed methods of improving the resistance of high strength steels to plastic fracture.

Influence of lyophilization factors and gelatin concentration on pore structures of atelocollagen/gelatin sponge biomaterial.

PubMed

Yang, Longqiang; Tanabe, Koji; Miura, Tadashi; Yoshinari, Masao; Takemoto, Shinji; Shintani, Seikou; Kasahara, Masataka

2017-07-26

This study aimed to investigate influences of lyophilization factors and gelatin concentration on pore structures of ACG sponge. ACG sponges of different freezing temperatures (-30, -80 and -196 o C), freezing times (1, 2 and 24 h), gelatin concentrations (0.6%AC+0.15%G, 0.6%AC+0.6%G and 0.6%AC+2.4%G), and with 500 μM fluvastatin were fabricated. Pore structures including porosity and pore size were analyzed by scanning electron microscopy and ImageJ. The cytotoxic effects of ACG sponges were evaluated in vitro. Freezing temperature did not affect porosity while high freezing temperature (-30 o C) increased pore size. The high gelatin concentration group (0.6%AC+2.4%G) had decreased porosity and pore size. Freezing time and 500 μM fluvastatin did not affect pore structures. The cytotoxicity and cell proliferation assays revealed that ACG sponges had no cytotoxic effects on human mesenchymal stromal cell growth and proliferation. These results indicate that ACG sponge may be a good biomaterial scaffold for bone regeneration.

The sea urchin egg jelly coat is a three-dimensional fibrous network as seen by intermediate voltage electron microscopy and deep etching analysis.

PubMed

Bonnell, B S; Larabell, C; Chandler, D E

1993-06-01

The egg jelly (EJ) coat which surrounds the unfertilized sea urchin egg undergoes extensive swelling upon contact with sea water, forming a three-dimensional network of interconnected fibers extending nearly 50 microns from the egg surface. Owing to its solubility, this coat has been difficult to visualize by light and electron microscopy. However, Lytechinus pictus EJ coats remain intact, if the fixation medium is maintained at pH 9. The addition of alcian blue during the final dehydration step of sample preparation stains the EJ for visualization of resin embedded eggs by both light and electron microscopy. Stereo pairs taken of thick sections prepared for intermediate voltage electron microscopy (IVEM) produce a three-dimensional image of the EJ network, consisting of interconnected fibers decorated along their length by globular jelly components. Using scanning electron microscopy (SEM), we have shown that before swelling, EJ exists in a tightly bound network of jelly fibers, 50-60 nm in diameter. In contrast, swollen EJ consists of a greatly extended network whose fibrous components measure 10 to 30 nm in diameter. High resolution stereo images of hydrated jelly produced by the quick-freeze/deep-etch/rotary-shadowing technique (QF/DE/RS) show nearly identical EJ networks, suggesting that dehydration does not markedly alter the structure of this extracellular matrix.

Thermal Destabilization of Collagen Matrix Hierarchical Structure by Freeze/Thaw

PubMed Central

Ozcelikkale, Altug; Han, Bumsoo

2016-01-01

This study aims to characterize and understand the effects of freezing on collagen structures and functionality. Specifically, thermodynamic destabilization of collagen at molecular- and fibril-levels by combination of low temperatures and freezing were experimentally characterized using modulated differential scanning calorimetry. In order to delineate the effects of sub-zero temperature and water-ice phase change, we hypothesized that the extent of destabilization can be determined based on post-thaw heat induced thermal denaturation of collagen. It is found that thermal denaturation temperature of collagen in hydrogel decreases by 1.4–1.6°C after freeze/thaw while no such decrease is observed in the case of molecular solution. The destabilization is predominantly due to ice formation. Exposure to low temperatures in the absence of ice has only minimal effect. Calorimetry measurements combined with morphological examination of collagen matrices by scanning electron microscopy suggest that freezing results in destabilization of collagen fibrils due to expansion of intrafibrillar space by ice formation. This fibril-level damage can be alleviated by use of cryoprotectant DMSO at concentrations as low as 0.5 M. A theoretical model explaining the change in collagen post-thaw thermal stability by freezing-induced fibril expansion is also proposed. PMID:26765741

Hot Ductility Behavior of an 8 Pct Cr Roller Steel

NASA Astrophysics Data System (ADS)

Wang, Zhenhua; Sun, Shuhua; Shi, Zhongping; Wang, Bo; Fu, Wantang

2015-04-01

The hot ductility of an 8 pct Cr roller steel was determined between 1173 K and 1473 K (900 °C and 1200 °C) at strain rates of 0.01 to 10 s-1 through tensile testing. The fracture morphology was observed using scanning electron microscopy, and the microstructure was examined through optical microscopy and transmission electron microscopy. The dependence of the hot ductility behavior on the deformation conditions, grain size, and precipitation was analyzed. The relationship between the reduction in area and the natural logarithm of the Zener-Hollomon parameter (ln Z) was found to be a second-order polynomial. When ln Z was greater than 40 s-1, the hot ductility was poor and fracture was mainly caused by incompatible deformation between the grains. When ln Z was between 32 and 40 s-1, the hot ductility was excellent and the main fracture mechanism was void linking. When ln Z was below 32 s-1, the hot ductility was poor and fracture was mainly caused by grain boundary sliding. A fine grain structure is beneficial for homogenous deformation and dynamic recrystallization, which induces better hot ductility. The effect of M7C3 carbide particles dispersed in the matrix on the hot ductility was small. The grain growth kinetics in the 8 pct Cr steel were obtained between 1373 K and 1473 K (1100 °C and 1200 °C). Finally, optimized preheating and forging procedures for 8 pct Cr steel rollers are provided.

Application of a quick-freezing and deep-etching method to pathological diagnosis: a case of elastofibroma.

PubMed

Hemmi, Akihiro; Tabata, Masahiko; Homma, Taku; Ohno, Nobuhiko; Terada, Nobuo; Fujii, Yasuhisa; Ohno, Shinichi; Nemoto, Norimichi

2006-04-01

A case of elastofibroma in a middle-aged Japanese woman was examined by the quick-freezing and deep-etching (QF-DE) method, as well as by immunohistochemistry and conventional electron microscopy. The slowly growing tumor developed at the right scapular region and was composed of fibrous connective tissue with unique elastic materials called elastofibroma fibers. A normal elastic fiber consists of a central core and peripheral zone, in which the latter has small aggregates of 10 nm microfibrils. By the QF-DE method, globular structures consisting of numerous fibrils (5-20 nm in width) were observed between the collagen bundles. We could confirm that they were microfibril-rich peripheral zones of elastofibroma fibers by comparing the replica membrane and conventional electron microscopy. One of the characteristics of elastofibroma fibers is that they are assumed to contain numerous microfibrils. Immunohistochemically, spindle tumor cells showed positive immunoreaction for vimentin, whereas alpha-smooth muscle actin, desmin, S-100 protein and CD34 showed negative immunoreaction. By conventional electron microscopy, the tumor cell had thin cytoplasmic processes, pinocytotic vesicles and prominent rough endoplasmic reticulum. Abundant intracytoplasmic filaments were observed in some tumor cells. Thick lamina-like structures along with their inner nuclear membrane were often observed in the tumor cell nuclei. The whole image of the tumor cell was considered to be a periosteal-derived cell, which would produce numerous microfibrils in the peripheral zone of elastofibroma fibers. This study indicated that the QF-DE method could be applied to the pathological diagnosis and analysis of pathomechanism, even for surgical specimens obtained from a patient.

Distribution of coniferin in freeze-fixed stem of Ginkgo biloba L. by cryo-TOF-SIMS/SEM

NASA Astrophysics Data System (ADS)

Aoki, Dan; Hanaya, Yuto; Akita, Takuya; Matsushita, Yasuyuki; Yoshida, Masato; Kuroda, Katsushi; Yagami, Sachie; Takama, Ruka; Fukushima, Kazuhiko

2016-08-01

To clarify the role of coniferin in planta, semi-quantitative cellular distribution of coniferin in quick-frozen Ginkgo biloba L. (ginkgo) was visualized by cryo time-of-flight secondary ion mass spectrometry and scanning electron microscopy (cryo-TOF-SIMS/SEM) analysis. The amount and rough distribution of coniferin were confirmed through quantitative chromatography measurement using serial tangential sections of the freeze-fixed ginkgo stem. The lignification stage of the sample was estimated using microscopic observations. Coniferin distribution visualized at the transverse and radial surfaces of freeze-fixed ginkgo stem suggested that coniferin is stored in the vacuoles, and showed good agreement with the assimilation timing of coniferin to lignin in differentiating xylem. Consequently, it is suggested that coniferin is stored in the tracheid cells of differentiating xylem and is a lignin precursor.

Light and electron microscopy study of the spermatheca of Eupholidoptera chabrieri bimucronata (Ramme, 1927) and Uromenus brevicollis trinacriae La Greca 1964 (Orthoptera: Tettigoniidae).

PubMed

Viscuso, Renata; Camiolo, Giuseppina; Vitale, Danilo G M

2015-07-01

A study by both optical and electron microscopy has been carried out on the spermatheca of Eupholidoptera chabrieri bimucronata and Uromenus brevicollis trinacriae (Orthoptera: Tettigoniidae). In both the examined species, the spermatheca consists of a sac/kidney-shaped seminal receptacle and a more or less tortuous spermathecal duct that opens into the common oviduct. The wall of both the organs consists of a pseudostratified epithelium surmounted by a cuticular intima; the latter is made up of a thicker endocuticle and an epicuticle. The epithelium shows two different cell types, irregularly arranged and with well differentiated functions: cuticle-forming and gland cells. In both the species, the cuticle-forming cells perform other functions, in addition to producing the cuticular intima. The gland cells never come in contact with the cuticular intima, have inside the reservoir a secretion whose appearance can diversify also in contiguous zones of the seminal receptacle. Based on our findings in both the species, the functions of the seminal receptacle would differ from those of the spermathecal duct. In the latter, some areas of the wall of the connecting tract show an activity of lysis, by contiguous epithelial cells, that could play a role in control and selection of spermatozoa. As for the feather-shaped spermatodesms, similar in both the species, freeze-fracture observations have shown that the acrosome of each spermatozoon regularly covers three-quarters of the extension of the acrosome of the following spermatozoon. Finally, the significance of our findings, compared with what is known in literature, is discussed. © 2015 Wiley Periodicals, Inc.

Revisiting dirt cracking as a physical weathering process in warm deserts

NASA Astrophysics Data System (ADS)

Dorn, Ronald I.

2011-12-01

A half century ago C.D. Ollier proposed that insolation-driven temperature changes expand and contract fill in fissures enough to widen cracks, a process that would permit progressively deeper penetration of fissure fills, that would in turn generate a positive feedback of greater and greater strain until desert boulders and bedrock shatters. Although desert physical weathering by "dirt cracking" has occasionally been cited, this hypothesized process remains without support from subsequent research. Here, field observations, electron microscopy, X-ray powder diffraction, particle-size analysis, and laboratory experiments shed new light on dirt cracking. Little clear evidence supports the original notion of expansive pressures from thermal fluctuations. However, mineralogical, high resolution transmission electron microscopy, back-scattered electron microscopy, and experimental evidence support two alternative processes of widening fractures: wetting and drying of fills inside fissures; and the precipitation and remobilization of calcium carbonate. A re-envisioned dirt-cracking wedging process starts with calcium carbonate precipitating in fissures less than 5 μm wide. First precipitation, and then ongoing dissolution of this laminar calcrete, opens enough space for dust to penetrate into these narrow fractures. Wetting of expansive clays in the fissure fill exerts enough pressure to widen and deepen the fissure, allowing the carbonate precipitation process to penetrate even deeper and allowing even more dust to move into a fracture. As the dust infiltrates, its texture changes from a chaotic mix of particles to an alignment of clays parallel to fissure sides. This parallel alignment could increase the efficiency of fill wedging. Ollier's concept of a positive feedback remains supported; each increment of fracture deepening and widening permits more, even deeper infiltration of laminar calcrete and dust. Field and electron microscope observations of rock spalling in the winter of 2010 are consistent with Ollier's hypothesis that dirt cracking is a common physical weathering process in deserts that splits rocks of all different sizes.

Measurement of Total Calcium in Neurons by Electron Probe X-ray Microanalysis

PubMed Central

Pivovarova, Natalia B.; Andrews, S. Brian

2013-01-01

In this article the tools, techniques, and instruments appropriate for quantitative measurements of intracellular elemental content using the technique known as electron probe microanalysis (EPMA) are described. Intramitochondrial calcium is a particular focus because of the critical role that mitochondrial calcium overload plays in neurodegenerative diseases. The method is based on the analysis of X-rays generated in an electron microscope (EM) by interaction of an electron beam with the specimen. In order to maintain the native distribution of diffusible elements in electron microscopy specimens, EPMA requires "cryofixation" of tissue followed by the preparation of ultrathin cryosections. Rapid freezing of cultured cells or organotypic slice cultures is carried out by plunge freezing in liquid ethane or by slam freezing against a cold metal block, respectively. Cryosections nominally 80 nm thick are cut dry with a diamond knife at ca. -160 °C, mounted on carbon/pioloform-coated copper grids, and cryotransferred into a cryo-EM using a specialized cryospecimen holder. After visual survey and location mapping at ≤-160 °C and low electron dose, frozen-hydrated cryosections are freeze-dried at -100 °C for ~30 min. Organelle-level images of dried cryosections are recorded, also at low dose, by means of a slow-scan CCD camera and subcellular regions of interest selected for analysis. X-rays emitted from ROIs by a stationary, focused, high-intensity electron probe are collected by an energy-dispersive X-ray (EDX) spectrometer, processed by associated electronics, and presented as an X-ray spectrum, that is, a plot of X-ray intensity vs. energy. Additional software facilitates: 1) identification of elemental components by their "characteristic" peak energies and fingerprint; and 2) quantitative analysis by extraction of peak areas/background. This paper concludes with two examples that illustrate typical EPMA applications, one in which mitochondrial calcium analysis provided critical insight into mechanisms of excitotoxic injury and another that revealed the basis of ischemia resistance. PMID:24300079

Physicochemical properties and bioactivity of freeze-cast chitosan nanocomposite scaffolds reinforced with bioactive glass.

PubMed

Pourhaghgouy, Masoud; Zamanian, Ali; Shahrezaee, Mostafa; Masouleh, Milad Pourbaghi

2016-01-01

Chitosan based nanocomposite scaffolds were prepared by freeze casting method through blending constant chitosan concentration with different portions of synthesized bioactive glass nanoparticles (BGNPs). Transmission Electron Microscopy (TEM) image showed that the particles size of bioactive glass (64SiO2.28CaO.8P2O5) prepared by sol-gel method was approximately less than 20 nm. Fourier Transform Infrared Spectroscopy (FT-IR) and X-ray Diffraction (XRD) analysis showed proper interfacial bonding between BGNPs and chitosan polymers. Scanning Electron Microscopy (SEM) images depicted a unidirectional structure with homogenous distribution of BGNPs among chitosan matrix associated with the absence of pure chitosan scaffold's wall pores after addition of only 10 wt.% BGNPs. As the BGNP content increased from 0 to 50 wt.%, the compressive strength and compressive module values increased from 0.034 to 0.419 MPa and 0.41 to 10.77 MPa, respectively. Biodegradation study showed that increase in BGNP content leads to growth of weight loss amount. The in vitro biomineralization studies confirmed the bioactive nature of all nanocomposites. Amount of 30 wt.% BGNPs represented the best concentration for absorption capacity and bioactivity behaviors. Copyright © 2015. Published by Elsevier B.V.

Cryopreservation of boar semen. III: Ultrastructure of boar spermatozoa frozen ultra-rapidly at various stages of conventional freezing and thawing.

PubMed

Bwanga, C O; Ekwall, H; Rodriguez-Martinez, H

1991-01-01

Ejaculated boar spermatozoa subjected to a conventional freezing and thawing process, were ultra-rapidly fixed, freeze-substituted and examined by electron microscopy to monitor the presence of real or potential intracellular ice and the degree of cell protection attained with the different extenders used during the process. Numerous ice crystal marks representing the degree of hydration of the cells were located in the perinuclear space of those spermatozoa not in proper contact with the extender containing glycerol (i.e. prior to freezing). The spermatozoa which were in proper contact with the extenders presented a high degree of preservation of the acrosomes, plasma membranes as well as the nuclear envelopes. No ice marks were detected in acrosomes before thawing, indicating that the conventional assayed cryopreservation method provided a good protection against cryoinjury. The presence of acrosomal changes (internal vesiculization, hydration and swelling) in thawed samples however, raises serious questions about the thawing procedure employed.

The Freezing Bomb

ERIC Educational Resources Information Center

Mills, Allan

2010-01-01

The extreme pressures that are generated when water freezes were traditionally demonstrated by sealing a small volume in a massive cast iron "bomb" and then surrounding it with a freezing mixture of ice and salt. This vessel would dramatically fail by brittle fracture, but no quantitative measurement of bursting pressure was available. Calculation…

Improving the mechanical properties of nano-hydroxyapatite

NASA Astrophysics Data System (ADS)

Khanal, Suraj Prasad

Hydroxyapatite (HAp) is an ideal bioactive material that is used in orthopedics. Chemical composition and crystal structure properties of HAp are similar to the natural bone hence it promotes bone growth. However, its mechanical properties of synthetic HAp are not sufficient for major load-bearing bone replacement. The potential of improving the mechanical properties of synthetic hydroxyapatite (HAp) by incorporating carboxyl functionalized single walled carbon nanotubes (CfSWCNT) and polymerized epsilon-caprolactam (nylon) is studied. The fracture toughness, tensile strength, Young's modulus, stiffness and fracture energy were studied for a series of HAp samples with CfSWCNT concentrations varying from 0 to 1.5 wt. % without, and with nylon addition. X-ray diffraction (XRD), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and Differential Scanning Calorimetry (DSC) were used to characterize the samples. The fracture toughness and tensile test was performed under the standard protocol of ASTM D5045 and ASTM D638-02a respectively. Reproducible maximum values of (3.60 +/- 0.3) MPa.m1/2 for fracture toughness and 65.38 MPa for tensile strength were measured for samples containing 1 wt. % CfSWCNT and nylon. The Young's modulus, stiffness and fracture energy of the samples are 10.65 GPa, 1482.12 N/mm, and 644 J/m2 respectively. These values are comparable to those of the cortical bone. Further increase of the CfSWCNT content results to a decreased fracture toughness and tensile strength and formation of a secondary phase.

QEMSCAN° (Quantitative Evaluation of Minerals by Scanning Electron Microscopy): capability and application to fracture characterization in geothermal systems

NASA Astrophysics Data System (ADS)

Ayling, B.; Rose, P. E.; Zemach, E.; Drakos, P. S.; Petty, S.

2011-12-01

Fractures are important conduits for fluids in geothermal systems, and the creation and maintenance of fracture permeability is a fundamental aspect of EGS (Engineered Geothermal System) development. Hydraulic or chemical stimulation techniques are often employed to achieve this. In the case of chemical stimulation, an understanding of the minerals present in the fractures themselves is desirable to better design a stimulation effort (i.e. which chemical to use and how much). Borehole televiewer surveys provide important information about regional and local stress regimes and fracture characteristics (e.g. fracture aperture), and XRD is useful for examining bulk rock mineralogy, but neither technique is able to quantify the distribution of these minerals in fractures. QEMSCAN° is a fully-automated micro-analysis system that enables quantitative chemical analysis of materials and generation of high-resolution mineral maps and images as well as porosity structure. It uses a scanning electron microscopy platform (SEM) with an electron beam source in combination with four energy-dispersive X-ray spectrometers (EDS). The measured backscattered electron and electron-induced secondary X-ray emission spectra are used to classify sample mineralogy. Initial applications of QEMSCAN° technology were predominantly in the minerals industry and application to geothermal problems has remained limited to date. In this pilot study, the potential application of QEMSCAN° technology to fracture characterization was evaluated using samples of representative mineralized fractures in two geothermal systems (Newberry Volcano, Oregon and Brady's geothermal field, Nevada). QEMSCAN° results were compared with XRD and petrographic techniques. Nine samples were analyzed from each field, collected from the drill core in the 1000-1500 m depth range in two shallow wells (GEO-N2 at Newberry Volcano and BCH-3 at Brady's). The samples were prepared as polished thin sections for QEMSCAN° analysis. Results indicate that a sampling resolution of 10 μm is sufficient to resolve fracture morphology and mineral zonation (where multiple episodes of mineralization occurred), and enables relatively fast data acquisition (3 cm2 can be analyzed in approximately 3 hours). Finer resolutions (down to 2.5 μm) take significantly longer, but can be used to provide additional spatial detail in areas of interest after a low resolution (10 μm) scan. Use of XRD data in conjunction with QEMSCAN° data is sometimes needed to distinguish geothermal alteration minerals with similar chemical compositions (clay minerals, micas and chlorite), however overall the technique appears to have excellent potential for geothermal applications.

Characterizing the composition of bone formed during fracture healing using scanning electron microscopy techniques.

PubMed

Perdikouri, Christina; Tägil, Magnus; Isaksson, Hanna

2015-01-01

About 5-10% of all bone fractures suffer from delayed healing, which may lead to non-union. Bone morphogenetic proteins (BMPs) can be used to induce differentiation of osteoblasts and enhance the formation of the bony callus, and bisphosphonates help to retain the newly formed callus. The aim of this study was to investigate if scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS) can identify differences in the mineral composition of the newly formed bone compared to cortical bone from a non-fractured control. Moreover, we investigate whether the use of BMPs and bisphosphonates-alone or combined-may have an effect on bone mineralization and composition. Twelve male Sprague-Dawley rats at 9 weeks of age were randomly divided into four groups and treated with (A) saline, (B) BMP-7, (C) bisphosphonates (Zoledronate), and (D) BMP-7 + Zoledronate. The rats were sacrificed after 6 weeks. All samples were imaged using SEM and chemically analyzed with EDS to quantify the amount of C, N, Ca, P, O, Na, and Mg. The Ca/P ratio was the primary outcome. In the fractured samples, two areas of interest were chosen for chemical analysis with EDS: the callus and the cortical bone. In the non-fractured samples, only the cortex was analyzed. Our results showed that the element composition varied to a small extent between the callus and the cortical bone in the fractured bones. However, the Ca/P ratio did not differ significantly, suggesting that the mineralization at all sites is similar 6 weeks post-fracture in this rat model.

Micro-fractures produced in the Cadalso de los Vidrios granite (Madrid) subjected to Freeze-Thaw Durability Testing

NASA Astrophysics Data System (ADS)

Freire-Lista, D. M.; Varas-Muriel, M. J.; Fort, R.

2012-04-01

A specific leucogranite (fine to medium grain sized) from Cadalso de los Vidrios, Madrid, Spain, from where it takes the name of the stone variety, which is a traditional heritage building material used in Central Spain, was subjected to freezing-thaw durability tests or accelerated artificial ageing tests (according to Spanish standard EN 12371:2001) to assess its durability by means of ultrasonic velocity measurements (a non-destructive technique), and optical and fluorescence petrography using a polarized optical microscope (destructive technique), both techniques used before, during and after laboratory ageing tests, or in other words, what is determined is the improvement or deterioration in some properties. The measurement of the ultrasonic velocity in the leucogranite cubic test specimens along the freezing-thaw cycles shown that the velocity diminishes with the number of cycles, in relation to the decay that the stones were experiencing. This deterioration can be observed by the loss of crystalline minerals in the surface of the analyzed samples and by the micro-fractures appearance up to one centimeter deep, which have been detected by the petrographic techniques previously mentioned. The images taken by means of the fluorescence microscope clearly show the micro-fractures generated during the durability test. These images have been processed and analyzed by the UTHSCSA Image Tool program with the purpose of being able to quantify the degree of decay that this type of crystalline materials undergone, when subjected to a number of freezing-thaw test cycles. It is therefore an effective, reliable and complementary technique to that of the petrography analysis, both optical and fluorescence ones. In the first cycles of the ageing test, the micro-fractures propagate along crystals edges and during the last cycles of the test, intracrystalline micro-fractures are generated, which are developed in different ways depending on the mineralogy of the crystals. Thus, the quartz crystals are those that undergo more intracrystalline micro-fractures, whereas the biotites, behave in a more ductile form and they are not micro-fractured. Both analytical techniques give information of this granite deterioration, showing a relation between the number of freezing-thaw cycles, the superficial micro-fractures proliferation and the decrease of ultrasonic waves propagation velocity produced by the ageing cycles.

Ultrastructure of the extracellular matrix of bovine dura mater, optic nerve sheath and sclera.

PubMed

Raspanti, M; Marchini, M; Della Pasqua, V; Strocchi, R; Ruggeri, A

1992-10-01

The sclera, the outermost sheath of the optic nerve and the dura mater have been investigated histologically and ultrastructurally. Although these tissues appear very similar under the light microscope, being dense connective tissues mainly composed of collagen bundles and a limited amount of cells and elastic fibres, they exhibit subtle differences on electron microscopy. In the dura and sclera collagen appears in the form of large, nonuniform fibrils, similar to those commonly found in tendons, while in the optic nerve sheath the fibrils appear smaller and uniform, similar to those commonly observed in reticular tissues, vessel walls and skin. Freeze-fracture also reveals these fibrils to have different subfibrillar architectures, straight or helical, which correspond to 2 distinct forms of collagen fibril previously described (Raspanti et al. 1989). The other extracellular matrix components also vary with the particular collagen fibril structure. Despite their common embryological derivation, the dura mater, optic nerve sheath and sclera exhibit diversification of their extracellular matrix consistent with the mechanical loads to which these tissues are subjected. Our observations indicate that the outermost sheath of the optic nerve resembles the epineurium of peripheral nerves rather than the dura to which it is commonly likened.

Morphological transitions of brain sphingomyelin are determined by the hydration protocol: ripples re-arrange in plane, and sponge-like networks disintegrate into small vesicles.

PubMed

Meyer, H W; Bunjes, H; Ulrich, A S

1999-06-01

The phase transition of hydrated brain sphingomyelin occurs at around 35 degrees C, which is close to the physiological temperature. Freeze-fracture electron microscopy is used to characterize different gel state morphologies in terms of solid-ordered and liquid-ordered phase states, according to the occurrence of ripples and other higher-dimensional bilayer deformations. Evidently, the natural mixed-chain sphingomyelin does not assume the flat L beta, phase but instead the rippled P beta, phase, with symmetric and asymmetric ripples as well as macroripples and an egg-carton pattern, depending on the incubation conditions. An unexpected difference was observed between samples that are hydrated above and below the phase transition temperature. When the lipid is hydrated at low temperature, a sponge-like network of bilayers is formed in the gel state, next to some normal lamellae. The network loses its ripples during cold-incubation, which indicates the formation of a liquid-ordered (lo) gel phase. Ripples re-appear upon warming and the sponge-like network disintegrates spontaneously and irreversibly into small vesicles above the phase transition.

Biophysical characterization of an integrin-targeted lipopolyplex gene delivery vector.

PubMed

Mustapa, M Firouz Mohd; Bell, Paul C; Hurley, Christopher A; Nicol, Alastair; Guénin, Erwann; Sarkar, Supti; Writer, Michele J; Barker, Susie E; Wong, John B; Pilkington-Miksa, Michael A; Papahadjopoulos-Sternberg, Brigitte; Shamlou, Parviz Ayazi; Hailes, Helen C; Hart, Stephen L; Zicha, Daniel; Tabor, Alethea B

2007-11-13

Nonviral gene delivery vectors now show good therapeutic potential: however, detailed characterization of the composition and macromolecular organization of such particles remains a challenge. This paper describes experiments to elucidate the structure of a ternary, targeted, lipopolyplex synthetic vector, the LID complex. This consists of a lipid component, Lipofectin (L) (1:1 DOTMA:DOPE), plasmid DNA (D), and a dual-function, cationic peptide component (I) containing DNA condensation and integrin-targeting sequences. Fluorophore-labeled lipid, peptide, and DNA components were used to formulate the vector, and the stoichiometry of the particles was established by fluorescence correlation spectroscopy (FCS). The size of the complex was measured by FCS, and the sizes of LID, L, LD, and ID complexes were measured by dynamic light scattering (DLS). Fluorescence quenching experiments and freeze-fracture electron microscopy were then used to demonstrate the arrangement of the lipid, peptide, and DNA components within the complex. These experiments showed that the cationic portion of the peptide, I, interacts with the plasmid DNA, resulting in a tightly condensed DNA-peptide inner core; this is surrounded by a disordered lipid layer, from which the integrin-targeting sequence of the peptide partially protrudes.

Membrane-active mechanism of LFchimera against Burkholderia pseudomallei and Burkholderia thailandensis.

PubMed

Kanthawong, Sakawrat; Puknun, Aekkalak; Bolscher, Jan G M; Nazmi, Kamran; van Marle, Jan; de Soet, Johannes J; Veerman, Enno C I; Wongratanacheewin, Surasakdi; Taweechaisupapong, Suwimol

2014-10-01

LFchimera, a construct combining two antimicrobial domains of bovine lactoferrin, lactoferrampin265-284 and lactoferricin17-30, possesses strong bactericidal activity. As yet, no experimental evidence was presented to evaluate the mechanisms of LFchimera against Burkholderia isolates. In this study we analyzed the killing activity of LFchimera on the category B pathogen Burkholderia pseudomallei in comparison to the lesser virulent Burkholderia thailandensis often used as a model for the highly virulent B. pseudomallei. Killing kinetics showed that B. thailandensis E264 was more susceptible for LFchimera than B. pseudomallei 1026b. Interestingly the bactericidal activity of LFchimera appeared highly pH dependent; B. thailandensis killing was completely abolished at and below pH 6.4. FITC-labeled LFchimera caused a rapid accumulation within 15 min in the cytoplasm of both bacterial species. Moreover, freeze-fracture electron microscopy demonstrated extreme effects on the membrane morphology of both bacterial species within 1 h of incubation, accompanied by altered membrane permeability monitored as leakage of nucleotides. These data indicate that the mechanism of action of LFchimera is similar for both species and encompasses disruption of the plasma membrane and subsequently leakage of intracellular nucleotides leading to cell dead.

Electrostatic control of phospholipid polymorphism.

PubMed

Tarahovsky, Y S; Arsenault, A L; MacDonald, R C; McIntosh, T J; Epand, R M

2000-12-01

A regular progression of polymorphic phase behavior was observed for mixtures of the anionic phospholipid, cardiolipin, and the cationic phospholipid derivative, 1, 2-dioleoyl-sn-glycero-3-ethylphosphocholine. As revealed by freeze-fracture electron microscopy and small-angle x-ray diffraction, whereas the two lipids separately assume only lamellar phases, their mixtures exhibit a symmetrical (depending on charge ratio and not polarity) sequence of nonlamellar phases. The inverted hexagonal phase, H(II,) formed from equimolar mixtures of the two lipids, i.e., at net charge neutrality (charge ratio (CR((+/-))) = 1:1). When one type of lipid was in significant excess (CR((+/-)) = 2:1 or CR((+/-)) = 1:2), a bicontinuous cubic structure was observed. These cubic phases were very similar to those sometimes present in cellular organelles that contain cardiolipin. Increasing the excess of cationic or anionic charge to CR((+/-)) = 4:1 or CR((+/-)) = 1:4 led to the appearance of membrane bilayers with numerous interlamellar contacts, i.e., sponge structures. It is evident that interactions between cationic and anionic moieties can influence the packing of polar heads and hence control polymorphic phase transitions. The facile isothermal, polymorphic interconversion of these lipids may have important biological and technical implications.

High-resolution x-ray diffraction microscopy of specifically labeled yeast cells

PubMed Central

Nelson, Johanna; Huang, Xiaojing; Steinbrener, Jan; Shapiro, David; Kirz, Janos; Marchesini, Stefano; Neiman, Aaron M.; Turner, Joshua J.; Jacobsen, Chris

2010-01-01

X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11–13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the α-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane and freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy. PMID:20368463

High-resolution x-ray diffraction microscopy of specifically labeled yeast cells

DOE PAGES

Nelson, Johanna; Huang, Xiaojing; Steinbrener, Jan; ...

2010-04-20

X-ray diffraction microscopy complements other x-ray microscopy methods by being free of lens-imposed radiation dose and resolution limits, and it allows for high-resolution imaging of biological specimens too thick to be viewed by electron microscopy. We report here the highest resolution (11-13 nm) x-ray diffraction micrograph of biological specimens, and a demonstration of molecular-specific gold labeling at different depths within cells via through-focus propagation of the reconstructed wavefield. The lectin concanavalin A conjugated to colloidal gold particles was used to label the α-mannan sugar in the cell wall of the yeast Saccharomyces cerevisiae. Cells were plunge-frozen in liquid ethane andmore » freeze-dried, after which they were imaged whole using x-ray diffraction microscopy at 750 eV photon energy.« less

Microstructure of bentonite in iron ore green pellets.

PubMed

Bhuiyan, Iftekhar U; Mouzon, Johanne; Schröppel, Birgit; Kaech, Andres; Dobryden, Illia; Forsmo, Seija P E; Hedlund, Jonas

2014-02-01

Sodium-activated calcium bentonite is used as a binder in iron ore pellets and is known to increase strength of both wet and dry iron ore green pellets. In this article, the microstructure of bentonite in magnetite pellets is revealed for the first time using scanning electron microscopy. The microstructure of bentonite in wet and dry iron ore pellets, as well as in distilled water, was imaged by various imaging techniques (e.g., imaging at low voltage with monochromatic and decelerated beam or low loss backscattered electrons) and cryogenic methods (i.e., high pressure freezing and plunge freezing in liquid ethane). In wet iron ore green pellets, clay tactoids (stacks of parallel primary clay platelets) were very well dispersed and formed a voluminous network occupying the space available between mineral particles. When the pellet was dried, bentonite was drawn to the contact points between the particles and formed solid bridges, which impart strength to the solid compact.

Nano-chemo-mechanical signature of conventional oil-well cement systems: Effects of elevated temperature and curing time

DOE Office of Scientific and Technical Information (OSTI.GOV)

Krakowiak, Konrad J., E-mail: kjkrak@mit.edu; Thomas, Jeffrey J., E-mail: JThomas39@slb.com; Musso, Simone, E-mail: SMusso@slb.com

2015-01-15

With ever more challenging (T,p) environments for cementing applications in oil and gas wells, there is a need to identify the fundamental mechanisms of fracture resistant oil well cements. We report results from a multi-technique investigation of behavior and properties of API class G cement and silica-enriched cement systems subjected to hydrothermal curing from 30 °C to 200 °C; including electron probe microanalysis, X-ray diffraction, thermogravimetry analysis, electron microscopy, neutron scattering (SANS), and fracture scratch testing. The results provide a new insight into the link between system chemistry, micro-texture and micro-fracture toughness. We suggest that the strong correlation found betweenmore » chemically modulated specific surface and fracture resistance can explain the drop in fracture properties of neat oil-well cements at elevated temperatures; the fracture property enhancement in silica-rich cement systems, between 110° and 175 °C; and the drop in fracture properties of such systems through prolonged curing over 1 year at 200 °C.« less

Features of plastic deformation and fracture of dispersion-strengthened V–Cr–Zr–W alloy depending on temperature of tension

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ditenberg, Ivan A.; Grinyaev, Konstantin V.; Tyumentsev, Alexander N.

2015-10-27

Influence of tension temperature on features of plastic deformation and fracture of V–4.23Cr–1.69Zr–7.56W alloy was investigated by scanning and transmission electron microscopy. It is shown that temperature increase leads to activation of the recovery processes, which manifests in the coarsening of microstructure elements, reducing the dislocation density, relaxation of continuous misorientations.

Amyloglucosidase enzymatic reactivity inside lipid vesicles

PubMed Central

Li, Mian; Hanford, Michael J; Kim, Jin-Woo; Peeples, Tonya L

2007-01-01

Efficient functioning of enzymes inside liposomes would open new avenues for applications in biocatalysis and bioanalytical tools. In this study, the entrapment of amyloglucosidase (AMG) (EC 3.2.1.3) from Aspergillus niger into dipalmitoylphosphatidylcholine (DPPC) multilamellar vesicles (MLVs) and large unilamellar vesicles (LUVs) was investigated. Negative-stain, freeze-fracture, and cryo-transmission electron microscopy images verified vesicle formation in the presence of AMG. Vesicles with entrapped AMG were isolated from the solution by centrifugation, and vesicle lamellarity was identified using fluorescence laser confocal microscopy. The kinetics of starch hydrolysis by AMG was modeled for two different systems, free enzyme in aqueous solution and entrapped enzyme within vesicles in aqueous suspension. For the free enzyme system, intrinsic kinetics were described by a Michaelis-Menten kinetic model with product inhibition. The kinetic constants, Vmax and Km, were determined by initial velocity measurements, and Ki was obtained by fitting the model to experimental data of glucose concentration-time curves. Predicted concentration-time curves using these kinetic constants were in good agreement with experimental measurements. In the case of the vesicles, the time-dependence of product (glucose) formation was experimentally determined and simulated by considering the kinetic behavior of the enzyme and the permeation of substrate into the vesicle. Experimental results demonstrated that entrapped enzymes were much more stable than free enyzme. The entrapped enzyme could be recycled with retention of 60% activity after 3 cycles. These methodologies can be useful in evaluating other liposomal catalysis operations. PMID:18271982

Cryo-planing of frozen-hydrated samples using cryo triple ion gun milling (CryoTIGM™).

PubMed

Chang, Irene Y T; Joester, Derk

2015-12-01

Cryo-SEM is a high throughput technique for imaging biological ultrastructure in its most pristine state, i.e. without chemical fixation, embedding, or drying. Freeze fracture is routinely used to prepare internal surfaces for cryo-SEM imaging. However, the propagation of the fracture plane is highly dependent on sample properties, and the resulting surface frequently shows substantial topography, which can complicate image analysis and interpretation. We have developed a broad ion beam milling technique, called cryogenic triple ion gun milling (CryoTIGM™ ['krī-ə-,tīm]), for cryo-planing frozen-hydrated biological specimens. Comparing sample preparation by CryoTIGM™ and freeze fracture in three model systems, Baker's yeast, mouse liver tissue, and whole sea urchin embryos, we find that CryoTIGM™ yields very large (∼700,000 μm(2)) and smooth sections that present ultrastructural details at similar or better quality than freeze-fractured samples. A particular strength of CryoTIGM™ is the ability to section samples with hard-soft contrast such as brittle calcite (CaCO3) spicules in the sea urchin embryo. Copyright © 2015 Elsevier Inc. All rights reserved.

Mechanical behavior and failure analysis of prosthetic retaining screws after long-term use in vivo. Part 4: Failure analysis of 10 fractured retaining screws retrieved from three patients.

PubMed

Al Jabbari, Youssef S; Fournelle, Raymond; Ziebert, Gerald; Toth, Jeffrey; Iacopino, Anthony M

2008-04-01

The aim of this study was to perform a failure analysis on fractured prosthetic retaining screws after long-term use in vivo. Additionally, the study addresses the commonly asked question regarding whether complex repeated functional occlusal forces initiate fatigue-type cracks in prosthetic retaining screws. Ten fractured prosthetic retaining screws retrieved from three patients treated with fixed detachable hybrid prostheses were subjected to a failure analysis. In patients 1 and 2, the middle three retaining screws of the prostheses were found fractured at retrieval time after they had been in service for 20 and 19 months, respectively. In patient 3, the middle three and one of the posterior retaining screws were found to be fractured at retrieval after they had been in service for 18 months. Low power stereomicroscopy and high-power scanning electron microscopy (SEM) were performed to analyze the fractured surfaces of the retaining screws examining fatigue cracks in greater detail. Typical fatigue failure characterized by ratchet mark formation was revealed by light microscopy and SEM for all examined screws. Using low magnification light microscopy, ratchet marks were visible on the fracture surfaces of only two screws. SEM examination revealed all three classical stages of fatigue failure, and it was possible to see the ratchet marks on the fracture surfaces of all specimens, indicating a fatigue zone. The final catastrophic overload fracture appeared fibrous, indicating ductile fracture. The final overload ductile fracture surfaces showed equiaxed dimples, suggesting tensile overload in all examined screws except in two specimens that showed an elongated dimple pattern indicating shear/tearing overload forces. Fracture of prosthetic retaining screws in hybrid prostheses occurs mainly through a typical fatigue mode involving mostly the middle anterior three screws. Fatigue cracks can grow in more than one prosthetic retaining screw, leading to fracture before the patient or clinician determines that any problem exists.

Mechanisms of Clostridium tyrobutyricum removal through natural creaming of milk: A microscopy study.

PubMed

D'Incecco, P; Faoro, F; Silvetti, T; Schrader, K; Pellegrino, L

2015-08-01

Clostridium tyrobutyricum is the main spoilage agent of late blowing defect (LBD) in Grana Padano and Parmigiano-Reggiano cheeses; LBD is characterized by openings and holes and is sometimes accompanied by cracks and an undesirable flavor. Even a very few spores remaining in the cheese curd may cause LBD; thus, it is essential to eradicate them during milk natural creaming. By this process, most of the bacteria, somatic cells, and spores rise to the top of the milk, together with the fat globules, and are removed with the cream. Previous studies suggested that milk immunoglobulins mediate the interactions between fat globules and bacteria that occur upon creaming but no direct evidence for this has been found. Moreover, other physical chemical interactions could be involved; for example, physical entrapment of spores among globule clusters. To maximize the efficiency of the natural creaming step in removing Cl. tyrobutyricum, it is essential to understand the nature of spore-globule interactions. With this aim, raw milk was contaminated with spores of Cl. tyrobutyricum before going to creaming overnight at 8°C, after which spore and bacteria removal was >90%. The obtained cream was analyzed by light interference contrast and fluorescence microscopy and by transmission electron microscopy (TEM). Results showed that most of the vegetative cells and spores, which were stained with malachite green before addition to milk, adhered tightly to the surface of single fat globules, the membranes of which appeared heterogeneous when stained with the fluorescent dye DilC18(3)-DS. Using the same dye, we observed transient and persistent interactions among globules, with formation of clusters of different sizes and partial coalescence of adhering membranes. Transmission electron microscopy examination of replicates of freeze-fractured cream allowed us to observe tight adhesion of spores to fat globules. Ultrathin sections revealed that this adhesion is mediated by an amorphous, slightly electron-opaque material, sometimes granular in appearance. Bacteria also adhered to different fat globules, linking them together, which suggests that adhesion was strong enough to maintain a stable contact. Although we cannot exclude physical entrapment of bacteria among fat globule clusters, we show for the first time that most of the bacteria are adhered to fat globules by an electron-opaque material whose nature has yet to be determined. Immunoglobulins are certainly the best candidates for adhesion but other compounds may be involved. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Dynamic recrystallization and grain boundary migration in B2 FeAl

NASA Technical Reports Server (NTRS)

Baker, I.; Gaydosh, D. J.

1987-01-01

Transmission electron microscopy and optical microscopy were used to examine polycrystalline specimens of the B2-structured alloy FeAl strained under tension to fracture at elevated temperature. Strain-induced grain boundary migration was observed above 900 K and dynamic recrystallization was found at 1000 K and 1100 K. Little evidence of dynamic recovery was evident but some networks were formed at 1100 K.

Scanning transmission ion micro-tomography (STIM-T) of biological specimens.

PubMed

Schwertner, Micheal; Sakellariou, Arthur; Reinert, Tilo; Butz, Tilman

2006-05-01

Computed tomography (CT) was applied to sets of Scanning Transmission Ion Microscopy (STIM) projections recorded at the LIPSION ion beam laboratory (Leipzig) in order to visualize the 3D-mass distribution in several specimens. Examples for a test structure (copper grid) and for biological specimens (cartilage cells, cygospore) are shown. Scanning Transmission Micro-Tomography (STIM-T) at a resolution of 260 nm was demonstrated for the first time. Sub-micron features of the Cu-grid specimen were verified by scanning electron microscopy. The ion energy loss measured during a STIM-T experiment is related to the mass density of the specimen. Typically, biological specimens can be analysed without staining. Only shock freezing and freeze-drying is required to preserve the ultra-structure of the specimen. The radiation damage to the specimen during the experiment can be neglected. This is an advantage compared to other techniques like X-ray micro-tomography. At present, the spatial resolution is limited by beam position fluctuations and specimen vibrations.

STRUCTURE OF MEMBRANE HOLES IN OSMOTIC AND SAPONIN HEMOLYSIS

PubMed Central

Seeman, P.; Cheng, D.; Iles, G. H.

1973-01-01

Serial section electron microscopy of hemolysing erythrocytes (fixed at 12 s after the onset of osmotic hemolysis) revealed long slits and holes in the membrane, extending to around 1 µm in length. Many but not all of the slits and holes (about 100–1000 Å wide) were confluent with one another. Ferritin and colloidal gold (added after fixation) only permeated those cells containing membrane defects. No such large holes or slits were seen in saponin-treated erythrocytes, and the membrane was highly invaginated, giving the ghost a scalloped outline. Freeze-etch electron microscopy of saponin-treated membranes revealed 40–50 Å-wide pits in the extracellular surface of the membrane. If these pits represent regions from which cholesterol was extracted, then cholesterol is uniformly distributed over the entire erythrocyte membrane. PMID:4566525

Refixation of Osteochondral Fractures by an Ultrasound-Activated Pin System - An Ovine In Vivo Examination Using CT and Scanning Electron Microscope.

PubMed

H, Neumann; A P, Schulz; S, Breer; A, Unger; B, Kienast

2015-01-01

Osteochondral injuries, if not treated appropriately, often lead to severe osteoarthritis of the affected joint. Without refixation of the osteochondral fragment, human cartilage only repairs these defects imperfectly. All existing refixation systems for chondral defects have disadvantages, for instance bad MRI quality in the postoperative follow-up or low anchoring forces. To address the problem of reduced stability in resorbable implants, ultrasound-activated pins were developed. By ultrasound-activated melting of the tip of these implants a higher anchoring is assumed. Aim of the study was to investigate, if ultrasound-activated pins can provide a secure refixation of osteochondral fractures comparing to conventional screw and conventional, resorbable pin osteosynthesis. CT scans and scanning electron microscopy should proovegood refixation results with no further tissue damage by the melting of the ultrasound-activated pins in comparison to conventional osteosynthesis. Femoral osteochondral fragments in sheep were refixated with ultrasound-activated pins (SonicPin™), Ethipins(®) and screws (Asnis™). The quality of the refixated fragments was examined after three month of full weight bearing by CT scans and scanning electron microscopy of the cartilage surface. The CT examination found almost no statistically significant difference in the quality of refixation between the three different implants used. Concerning the CT morphology, ultrasound-activated pins demonstrated at least the same quality in refixation of osteochondral fragments as conventional resorbable pins or screws. The scanning electron microscopy showed no major surface damage by the three implants, especially any postulated cartilage damage induced by the heat of the ultrasound-activated pin. The screws protruded above the cartilage surface, which may affect the opposingtibial surface. Using CT scans and scanning electron microscopy, the SonicPin™, the Ethipin(®) and screws were at least equivalent in refixation quality of osteochondral fragments.

Additional freeze hardiness in wheat acquired by exposure to -3 degreesC is associated with extensive physiological, morphological, and molecular changes.

PubMed

Herman, Eliot M; Rotter, Kelsi; Premakumar, Ramaswamy; Elwinger, G; Bae, Hanhong; Bae, Rino; Ehler-King, Linda; Chen, Sixue; Livingston, David P

2006-01-01

Cold-acclimated plants acquire an additional 3-5 degrees C increase in freezing tolerance when exposed to -3 degrees C for 12-18 h before a freezing test (LT50) is applied. The -3 degrees C treatment replicates soil freezing that can occur in the days or weeks leading to overwintering by freezing-tolerant plants. This additional freezing tolerance is called subzero acclimation (SZA) to differentiate it from cold acclimation (CA) that is acquired at above-freezing temperatures. Using wheat as a model, results have been obtained indicating that SZA is accompanied by changes in physiology, cellular structure, the transcriptome, and the proteome. Using a variety of assays, including DNA arrays, reverse transcription-polymerase chain reaction (RT-PCR), 2D gels with mass spectroscopic identification of proteins, and electron microscopy, changes were observed to occur as a consequence of SZA and the acquisition of added freezing tolerance. In contrast to CA, SZA induced the movement of intracellular water to the extracellular space. Many unknown and stress-related genes were upregulated by SZA including some with obvious roles in SZA. Many genes related to photosynthesis and plastids were downregulated. Changes resulting from SZA often appeared to be a loss of rather than an appearance of new proteins. From a cytological perspective, SZA resulted in alterations of organelle structure including the Golgi. The results indicate that the enhanced freezing tolerance of SZA is correlated with a wide diversity of changes, indicating that the additional freezing tolerance is the result of complex biological processes.

Effect of cellulose nanocrystals (CNC) particle morphology on dispersion and rheological and mechanical properties of polypropylene/CNC nanocomposites.

PubMed

Khoshkava, Vahid; Kamal, Musa R

2014-06-11

Polypropylene (PP) nanocomposites containing spray-dried cellulose nanocrystals (CNC), freeze-dried CNC, and spray-freeze-dried CNC (CNCSFD) were prepared via melt mixing in an internal batch mixer. Polarized light, scanning electron, and atomic force microscopy showed significantly better dispersion of CNCSFD in PP/CNC nanocomposites compared with the spray-dried and freeze-dried CNCs. Rheological measurements, including linear and nonlinear viscoelastic tests, were performed on PP/CNC samples. The microscopy results were supported by small-amplitude oscillatory shear tests, which showed substantial rises in the magnitudes of key rheological parameters of PP samples containing CNCSFD. Steady-shear results revealed a strong shear thinning behavior of PP samples containing CNCSFD. Moreover, PP melts containing CNCSFD exhibited a yield stress. The magnitude of the yield stress and the degree of shear thinning behavior increased with CNCSFD concentration. It was found that CNCSFD agglomerates with a weblike structure were more effective in modifying the rheological properties. This effect was attributed to better dispersion of the agglomerates with the weblike structure. Dynamic mechanical analysis showed considerable improvement in the modulus of samples containing CNCSFD agglomerates. The percolation mechanical model with modified volume percolation threshold and filler network strength values and the Halpin-Kardos model were used to fit the experimental results.

Filament organization revealed in platinum replicas of freeze-dried cytoskeletons

PubMed Central

1980-01-01

This report presents the appearance of rapidly frozen, freeze-dried cytoskeletons that have been rotary replicated with platinum and viewed in the transmission electron microscope. The resolution of this method is sufficient to visualize individual filaments in the cytoskeleton and to discriminate among actin, microtubules, and intermediate filaments solely by their surface substructure. This identification has been confirmed by specific decoration with antibodies and selective extraction of individual filament types, and correlated with light microscope immunocytochemistry and gel electrophoresis patterns. The freeze-drying preserves a remarkable degree of three-dimensionality in the organization of these cytoskeletons. They look strikingly similar to the meshwork of strands or "microtrabeculae" seen in the cytoplasm of whole cells by high voltage electron microscopy, in that the filaments form a lattice of the same configutation and with the same proportions of open area as the microtrabeculae seen in whole cells. The major differences between these two views of the structural elements of the cytoplasmic matrix can be attributed to the effects of aldehyde fixation and dehydration. Freeze-dried cytoskeletons thus provide an opportunity to study--at high resolution and in the absence of problems caused by chemical fixation--the detailed organization of filaments in different regions of the cytoplasm and at different stages of cell development. In this report the pattern of actin and intermediate filament organization in various regions of fully spread mouse fibroblasts is described. PMID:6893451

Study of Aging-Induced Degradation of Fracture Resistance of Alloy 617 Toward High-Temperature Applications

NASA Astrophysics Data System (ADS)

Singh, Aditya Narayan; Moitra, A.; Bhaskar, Pragna; Sasikala, G.; Dasgupta, Arup; Bhaduri, A. K.

2017-07-01

For the Alloy 617, the effect of aging on the fracture energy degradation has been investigated after aging for different time periods at 1023 K (750 °C). A sharp reduction in impact energy (by 55 pct vis-à-vis the as-received material) after 1000 hours of aging, as evaluated from room-temperature Charpy impact tests, has been observed. Further aging up to 10,000 hours has led to a degradation of fracture energy up to 78 pct. Fractographic examinations using scanning electron microscopy (SEM) have revealed a change in fracture mode from fibrous-ductile for the un-aged material to intergranular mode for the aged one. The extent of intergranular fracture increases with the increasing aging time, indicating a tendency of the material to undergo grain boundary embrittlement over long-term aging. Analysis of the transmission electron microscopy (TEM) micrographs along with selected area diffraction (SAD) patterns for the samples aged at 10,000 hours revealed finely dispersed γ' precipitates of size 30 to 40 nm, rich in Al and Ti, along with extensive precipitation of M23C6 at the grain boundaries. In addition, the presence of Ni3Si of size in the range of 110 to 120 nm also has been noticed. The extensive precipitation of M23C6 at the grain boundaries have been considered as a major reason for aging-induced embrittlement of this material.

Correlating microscopy techniques and ToF-SIMS analysis of fully grown mammalian oocytes.

PubMed

Gulin, Alexander; Nadtochenko, Victor; Astafiev, Artyom; Pogorelova, Valentina; Rtimi, Sami; Pogorelov, Alexander

2016-06-20

The 2D-molecular thin film analysis protocol for fully grown mice oocytes is described using an innovative approach. Time-of-flight secondary ion mass spectrometry (ToF-SIMS), scanning electron microscopy (SEM), atomic force microscopy (AFM) and optical microscopy imaging were applied to the same mice oocyte section on the same sample holder. A freeze-dried mice oocyte was infiltrated into embedding media, e.g. Epon, and then was cut with a microtome and 2 μm thick sections were transferred onto an ITO coated conductive glass. Mammalian oocytes can contain "nucleolus-like body" (NLB) units and ToF-SIMS analysis was used to investigate the NLB composition. The ion-spatial distribution in the cell components was identified and compared with the images acquired by SEM, AFM and optical microscopy. This study presents a significant advancement in cell embryology, cell physiology and cancer-cell biochemistry.

Technical Advances in Intracellular Detection Using Immuno-Gold Particles: Simple Cryofixation with Metal Contact Quick Freezing.

PubMed

Song, Chihong; Lee, Ju Huck; Jun, Sangmi; Chung, Jeong Min; Hyun, Jaekyung; Jung, Hyun Suk

2016-05-01

The preparation of biological specimens using cryofixation techniques ensures excellent visibility of intracellular structures and preserves the antigenic sites of subcellular molecules. Hence, cryofixation is an effective method of preparing samples for analyses using antibodies conjugated to gold nanoparticles that are designed to detect the localization of specific target molecules within cells. However, cryofixation cannot be utilized easily because it requires expensive equipment and skilled technologists, resulting in a high level of expense for researchers. Here, we describe a simple technical approach to cryofixation that uses metal contact quick freezing followed by a modified freeze substitution technique and immuno-gold labeling electron microscopy. Micrograph images of cells prepared using this modified cryofixation method demonstrated its superiority over chemical fixation for high contrast visualization of the morphologies of cellular components and preservation of antigenicity for immuno-gold labeling. This report provides valuable technical information related to the advancement of metal contact quick freezing techniques, which can be used to visualize biomedical events of interest in an easy, simple, and rapid manner.

Solidification of liposomes by freeze-drying: the importance of incorporating gelatin as interior support on enhanced physical stability.

PubMed

Guan, Peipei; Lu, Yi; Qi, Jianping; Niu, Mengmeng; Lian, Ruyue; Wu, Wei

2015-01-30

The main purpose of this study was to investigate the effect of gelatin as interior support on the physical stability of freeze-dried liposomes. Anticancer agent paclitaxel (PTX) was selected as a model drug. Freeze-dried liposomes containing interior gelatin support (GLs) were prepared by thin-film dispersion/freeze-drying method. Several properties of the GLs, including entrapment efficiency, particle size and gelation temperature, were extensively characterized. Encapsulation efficiency of conventional liposomes (CLs) and liposomes containing lyoprotectants as interior support dropped to lower than 20% after reconstitution, while GLs still maintained an entrapment efficiency of over 84%. Scanning electron microscopy revealed well preserved liposomal structure of GLs after reconstitution. Meanwhile, the particle size and entrapment efficiency of GLs were also well preserved after reconstitution. In contrary, deformation of CLs and recrystallization of PTX were observed, as well as significant changes in particle size and entrapment efficiency. Taken together, interior gelatin support obviously enhanced the physical stability of liposomes against the lyophilization stress. Copyright © 2014 Elsevier B.V. All rights reserved.

Investigation of Mechanical Properties and Fracture Simulation of Solution-Treated AA 5754

NASA Astrophysics Data System (ADS)

Kumar, Pankaj; Singh, Akhilendra

2017-10-01

In this work, mechanical properties and fracture toughness of as-received and solution-treated aluminum alloy 5754 (AA 5754) are experimentally evaluated. Solution heat treatment of the alloy is performed at 530 °C for 2 h, and then, quenching is done in water. Yield strength, ultimate tensile strength, impact toughness, hardness, fatigue life, brittle fracture toughness (K_{Ic} ) and ductile fracture toughness (J_{Ic} ) are evaluated for as-received and solution-treated alloy. Extended finite element method has been used for the simulation of tensile and fracture behavior of material. Heaviside function and asymptotic crack tip enrichment functions are used for modelling of the crack in the geometry. Ramberg-Osgood material model coupled with fracture energy is used to simulate the crack propagation. Fracture surfaces obtained from various mechanical tests are characterized by scanning electron microscopy.

Mechanical and structural characterizations of gamma- and alpha-alumina nanofibers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vahtrus, Mikk; Umalas, Madis; Polyakov, Boris

2015-09-15

We investigate the applicability of alumina nanofibers as a potential reinforcement material in ceramic matrix compounds by comparing the mechanical properties of individual nanofibers before and after annealing at 1400 °C. Mechanical testing is performed inside a scanning electron microscope (SEM), which enables observation in real time of the deformation and fracture of the fibers under loading, thereby providing a close-up inspection of the freshly fractured area in vacuum. Improvement of both the Young's modulus and the breaking strength for annealed nanofibers is demonstrated. Mechanical testing is supplemented with the structural characterization of the fibers before and after annealing usingmore » SEM, transmission electron microscopy and X-ray diffraction methods. - Highlights: • Mechanical properties of individual alumina nanofibers were measured using in situ SEM cantilevered beam bending technique. • Improvement of mechanical properties of the alumina fibers after annealing at 1400 °C is demonstrated. • Formation of branched structures is demonstrated and their mechanical properties are studied. • XRD and electron microscopy were used for structural characterization of untreated and annealed nanofibers.« less

Photoprotection Conferred by Changes in Photosynthetic Protein Levels and Organization during Dehydration of a Homoiochlorophyllous Resurrection Plant1

PubMed Central

Charuvi, Dana; Nevo, Reinat; Shimoni, Eyal; Naveh, Leah; Zia, Ahmad; Adam, Zach; Farrant, Jill M.; Kirchhoff, Helmut; Reich, Ziv

2015-01-01

During desiccation, homoiochlorophyllous resurrection plants retain most of their photosynthetic apparatus, allowing them to resume photosynthetic activity quickly upon water availability. These plants rely on various mechanisms to prevent the formation of reactive oxygen species and/or protect their tissues from the damage they inflict. In this work, we addressed the issue of how homoiochlorophyllous resurrection plants deal with the problem of excessive excitation/electron pressures during dehydration using Craterostigma pumilum as a model plant. To investigate the alterations in the supramolecular organization of photosynthetic protein complexes, we examined cryoimmobilized, freeze-fractured leaf tissues using (cryo)scanning electron microscopy. These examinations revealed rearrangements of photosystem II (PSII) complexes, including a lowered density during moderate dehydration, consistent with a lower level of PSII proteins, as shown by biochemical analyses. The latter also showed a considerable decrease in the level of cytochrome f early during dehydration, suggesting that initial regulation of the inhibition of electron transport is achieved via the cytochrome b6f complex. Upon further dehydration, PSII complexes are observed to arrange into rows and semicrystalline arrays, which correlates with the significant accumulation of sucrose and the appearance of inverted hexagonal lipid phases within the membranes. As opposed to PSII and cytochrome f, the light-harvesting antenna complexes of PSII remain stable throughout the course of dehydration. Altogether, these results, along with photosynthetic activity measurements, suggest that the protection of retained photosynthetic components is achieved, at least in part, via the structural rearrangements of PSII and (likely) light-harvesting antenna complexes into a photochemically quenched state. PMID:25713340

Validation of the shake test for detecting freeze damage to adsorbed vaccines.

PubMed

Kartoglu, Umit; Ozgüler, Nejat Kenan; Wolfson, Lara J; Kurzatkowski, Wiesław

2010-08-01

To determine the validity of the shake test for detecting freeze damage in aluminium-based, adsorbed, freeze-sensitive vaccines. A double-blind crossover design was used to compare the performance of the shake test conducted by trained health-care workers (HCWs) with that of phase contrast microscopy as a "gold standard". A total of 475 vials of 8 different types of World Health Organization prequalified freeze-sensitive vaccines from 10 different manufacturers were used. Vaccines were kept at 5 degrees C. Selected numbers of vials from each type were then exposed to -25 degrees C and -2 degrees C for 24-hour periods. There was complete concordance between HCWs and phase-contrast microscopy in identifying freeze-damaged vials and non-frozen samples. Non-frozen samples showed a fine-grain structure under phase contrast microscopy, but freeze-damaged samples showed large conglomerates of massed precipitates with amorphous, crystalline, solid and needle-like structures. Particles in the non-frozen samples measured from 1 microm (vaccines against diphtheria-tetanus-pertussis; Haemophilus influenzae type b; hepatitis B; diphtheria-tetanus-pertussis-hepatitis B) to 20 microm (diphtheria and tetanus vaccines, alone or in combination). By contrast, aggregates in the freeze-damaged samples measured up to 700 microm (diphtheria-tetanus-pertussis) and 350 microm on average. The shake test had 100% sensitivity, 100% specificity and 100% positive predictive value in this study, which confirms its validity for detecting freeze damage to aluminium-based freeze-sensitive vaccines.

EXTRACTION OF FRACTURE-MECHANICS AND TRANSMISSION-ELECTRON-MICROSCOPY SAMPLES FROM TRITIUM-EXPOSED RESERVOIRS USING ELECTRIC-DISCHARGE MACHINING

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morgan, M; Ken Imrich, K; Michael Tosten, M

2006-08-31

The Enhanced Surveillance Campaign is funding a program to investigate tritium aging effects on the structural properties of tritium reservoir steels. The program is designed to investigate how the structural properties of reservoir steels change during tritium service and to examine the role of microstructure and reservoir manufacturing on tritium compatibility. New surveillance tests are also being developed that can better gauge the long-term effects of tritium and its radioactive decay product, helium-3, on the properties of reservoir steels. In order to conduct these investigations, three types of samples are needed from returned reservoirs: tensile, fracture mechanics, and transmission-electron microscopymore » (TEM). An earlier report demonstrated how the electric-discharge machining (EDM) technique can be used for cutting tensile samples from serial sections of a 3T reservoir and how yield strength, ultimate strength and elongation could be measured from those samples. In this report, EDM was used successfully to section sub-sized fracture-mechanics samples from the inner and outer walls of a 3T reservoir and TEM samples from serial sections of a 1M reservoir. This report fulfills the requirements for the FY06 Level 3 milestone, TSR 15.1 ''Cut Fracture-Mechanics Samples from Tritium-Exposed Reservoir'' and TSR 15.2 ''Cut Transmission-electron-microscopy foils from Tritium-Exposed Reservoir'' for the Enhance Surveillance Campaign (ESC). This was in support of ESC L2-1870 Milestone-''Provide aging and lifetime assessments of selected components and materials for multiple enduring stockpile systems''.« less

Fracture behavior of nano-scale rubber-modified epoxies

NASA Astrophysics Data System (ADS)

Bacigalupo, Lauren N.

The primary focus of the first portion of this study is to compare physical and mechanical properties of a model epoxy that has been toughened with one of three different types of rubber-based modifier: a traditional telechelic oligomer (phase separates into micro-size particles), a core-shell latex particle (preformed nano-scale particles) and a triblock copolymer (self-assembles into nano-scale particles). The effect of modifier content on the physical properties of the matrix was determined using several thermal analysis methods, which provided insight into any inherent alterations of the epoxy matrix. Although the primary objective is to study the role of particle size on the fracture toughness, stiffness and strength were also determined since these properties are often reduced in rubber-toughened epoxies. It was found that since the CSR- and SBM-modified epoxies are composed of less rubber, thermal and mechanical properties of the epoxy were better maintained. In order to better understand the fracture behavior and mechanisms of the three types of rubber particles utilized in this study, extensive microscopy analysis was conducted. Scanning transmission electron microscopy (STEM) was used to quantify the volume fraction of particles, transmission optical microscopy (TOM) was used to determine plastic damage zone size, and scanning electron microscopy (SEM) was used to assess void growth in the plastic zone after fracture. By quantifying these characteristics, it was then possible to model the plastic damage zone size as well as the fracture toughness to elucidate the behavior of the rubber-modified epoxies. It was found that localized shear yielding and matrix void growth are the active toughening mechanisms in all rubber-modified epoxies in this study, however, matrix void growth was more prevalent. The second portion of this study investigated the use of three acrylate-based triblocks and four acrylate-based diblocks to modify a model epoxy system. By varying block lengths and the polarity of the epoxy-miscible blocks, a variety of morphologies were generated (such as spherical micelles, layer particles and worm-like micelles). It was found that in some cases, the epoxy-miscible block did not yield domains substantial enough to facilitate increases in toughness. Overall, the thermal and mechanical properties of the acrylate-based triblock- and diblock-modified epoxies were found to be similar to CTBN-modified epoxy, which was used as a control. However, there were properties that were improved with the acrylate-based diblock-modified epoxies when compared to the acrylate-based triblock modified epoxies. Specifically, the viscosity penalty of the diblock-modified epoxies was shown to be a marked improvement over the triblock-modified epoxies, especially given that the fracture toughness values are similar. This reduction in the viscosity penalty becomes an important criterion when considering processing procedures and applications. Additionally, comparing the morphology of the resulting modified-epoxies utilizing atomic force microscopy (AFM) and scanning electron microscopy (SEM) led to a better understanding of the relationship between the particle morphology obtained and the physical properties of the acrylate-based rubber-modified epoxy systems in this research.

The application of polyethylene glycol (PEG) to electron microscopy

PubMed Central

1980-01-01

The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine- coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis. PMID:7400222

The application of polyethylene glycol (PEG) to electron microscopy.

PubMed

Wolosewick, J J

1980-08-01

The cytoplasm of cells from a variety of tissues has been viewed in sections (0.25-1 micrometers) devoid of any embedding resin. Glutaraldehyde- and osmium tetroxide-fixed tissues were infiltrated and embedded in a water-miscible wax, polyethylene glycol (PEG), and subsequently sectioned on dry glass or diamond knives. The PEG matrix was removed and the sections were placed on Formvarcarbon-polylysine-coated grids, dehydrated, dried by the critical-point method, and observed in either the high- or low-voltage electron microscope. Stereoscopic views of cells devoid of embedding resin present an image of cell utrastructure unobscured by electron-scattering resins similar to the image of whole, unembedded critical-point-dried or freeze-dried cultured cells observed by transmission electron microscopy. All organelles, including the cytoskeletal structures, are identified and appear not to have been damaged during processing, although membrane components appear somewhat less distinct. The absence of an embedding matrix eliminates the need for additional staining to increase contrast, unlike the situation with specimens embedded in standard electron-scattering resins. The PEG technique thus appears to be a valuable adjunct to conventional methods for ultrastructural analysis.

Importance of microscopy in durability studies of solidified and stabilized contaminated soils

USGS Publications Warehouse

Klich, I.; Wilding, L.P.; Drees, L.R.; Landa, E.R.

1999-01-01

Solidification/stabilization (S/S) is recognized by the U.S. EPA as a best demonstrated available technology for the containment of contaminated soils and other hazardous wastes that cannot be destroyed by chemical, thermal, or biological means. Despite the increased use of S/S technologies, little research has been conducted on the weathering and degradation of solidified and stabilized wastes once the treated materials have been buried. Published data to verify the performance and durability of landfilled treated wastes over time are rare. In this preliminary study, optical and electron microscopy (scanning electron microscopy [SEM], transmission electron microscopy [TEM] and electron probe microanalyses [EPMA]) were used to evaluate weathering features associated with metal-bearing contaminated soil that had been solidified and stabilized with Portland cement and subsequently buried on site, stored outdoors aboveground, or achieved in a laboratory warehouse for up to 6 yr. Physical and chemical alteration processes identified include: freeze-thaw cracking, cracking caused by the formation of expansive minerals such as ettringite, carbonation, and the movement of metals from waste aggregates into the cement micromass. Although the extent of degradation after 6 yr is considered slight to moderate, results of this study show that the same environmental concerns that affect the durability of concrete must be considered when evaluating the durability and permanence of the solidification and stabilization of contaminated soils with cement. In addition, such evaluations cannot be based on leaching and chemical analyses alone. The use of all levels of microscopic analyses must be incorporated into studies of the long-term performance of S/S technologies.Solidification/stabilization (S/S) is recognized by the U.S. EPA as a best demonstrated available technology for the containment of contaminated soils and other hazardous wastes that cannot be destroyed by chemical, thermal, or biological means. Despite the increased use of S/S technologies, little research has been conducted on the weathering and degradation of solidified and stabilized wastes once the treated materials have been buried. Published data to verify the performance and durability of landfilled treated wastes over time are rare. In this preliminary study, optical and electron microscopy (scanning electron microscopy [SEM], transmission electron microscopy [TEM] and electron probe microanalyses [EPMA]) were used to evaluate weathering features associated with metal-bearing contaminated soil that had been solidified and stabilized with Portland cement and subsequently buried on site, stored outdoors aboveground, or archived in a laboratory, warehouse for up to 6 yr. Physical and chemical alteration processes identified include: freeze-thaw cracking, cracking caused by the formation of expansive minerals such as ettringite, carbonation, and the movement of metals from waste aggregates into the cement micromass. Although the extent of degradation after 6 yr is considered slight to moderate, results of this study show that the same environmental concerns that affect the durability of concrete must be considered when evaluating the durability and permanence of the solidification and stabilization of contaminated soils with cement. In addition, such evaluations cannot be based on leaching and chemical analyses alone. The use of all levels of microscopic analyses must be incorporated into studies of the long-term performance of S/S technologies.

Comparison of the effects of glycerol, dimethyl sulfoxide, and hydroxyethyl starch solutions for cryopreservation of avian red blood cells.

PubMed

Graham, Jennifer E; Meola, Dawn M; Kini, Nisha R; Hoffman, Andrew M

2015-06-01

To compare effectiveness of glycerol, dimethyl sulfoxide (DMSO), and hydroxyethyl starch (HES) solutions for cryopreservation of avian RBCs. RBCs from 12 healthy Ameraucana hens (Gallus gallus domesticus). RBCs were stored in 20% (wt/vol) glycerol, 10% (wt/vol) DMSO freezing medium, or various concentrations of HES solution (7.5%, 11.5%, and 20% [wt/vol]) and frozen for 2 months in liquid nitrogen. Cells were then thawed and evaluated by use of cell recovery and saline stability tests, cell staining (7-aminoactinomycin D and annexin V) and flow cytometry, and scanning electron microscopy. Percentage of RBCs recovered was highest for 20% glycerol solution (mean ± SE, 99.71 ± 0.04%) and did not differ significantly from the value for 7.5% HES solution (99.57 ± 0.04%). Mean saline stability of RBCs was highest for 10% DMSO (96.11 ± 0.25%) and did not differ significantly from the value for 20% HES solution (95.74 ± 0.25%). Percentages of cells with 7-aminoactinomycin D staining but without annexin V staining (indicating necrosis or late apoptosis) were lowest for 10% DMSO freezing medium (3%) and 20% glycerol solution (1%) and highest for all HES concentrations (60% to 80%). Scanning electron microscopy revealed severe membrane changes in RBCs cryopreserved in 20% HES solution, compared with membrane appearance in freshly harvested RBCs and RBCs cryopreserved in 10% DMSO freezing medium. Cryopreservation of avian RBCs with HES solution, regardless of HES concentration, resulted in greater degrees of apoptosis and cell death than did cryopreservation with other media. Transfusion with RBCs cryopreserved in HES solution may result in posttransfusion hemolysis in birds.

Advantages of Environmental Scanning Electron Microscopy in Studies of Microorganisms

DTIC Science & Technology

1993-01-01

external structures, cultures of Aspergillus sp. and Cunninghamella sp. but details of cytoplasmic organization were lacking were maintained on corn meal ... agar . (Fig. 7). Uncoated, freeze-dried Euglena sp. cells exam- ined with either ESD or BSE detectors did not exhibit Specimen Preparation for ESEM...approximately Agar plugs from sporulating cultures of Cunning- 1.0 pLm in diameter (Fig. 23). Many of the vesicles ap- hamella sp., Mucor sp., and Aspergillus

Gap junctions on hippocampal mossy fiber axons demonstrated by thin-section electron microscopy and freeze–fracture replica immunogold labeling

PubMed Central

Hamzei-Sichani, Farid; Kamasawa, Naomi; Janssen, William G. M.; Yasumura, Thomas; Davidson, Kimberly G. V.; Hof, Patrick R.; Wearne, Susan L.; Stewart, Mark G.; Young, Steven R.; Whittington, Miles A.; Rash, John E.; Traub, Roger D.

2007-01-01

Gap junctions have been postulated to exist between the axons of excitatory cortical neurons based on electrophysiological, modeling, and dye-coupling data. Here, we provide ultrastructural evidence for axoaxonic gap junctions in dentate granule cells. Using combined confocal laser scanning microscopy, thin-section transmission electron microscopy, and grid-mapped freeze–fracture replica immunogold labeling, 10 close appositions revealing axoaxonic gap junctions (≈30–70 nm in diameter) were found between pairs of mossy fiber axons (≈100–200 nm in diameter) in the stratum lucidum of the CA3b field of the rat ventral hippocampus, and one axonal gap junction (≈100 connexons) was found on a mossy fiber axon in the CA3c field of the rat dorsal hippocampus. Immunogold labeling with two sizes of gold beads revealed that connexin36 was present in that axonal gap junction. These ultrastructural data support computer modeling and in vitro electrophysiological data suggesting that axoaxonic gap junctions play an important role in the generation of very fast (>70 Hz) network oscillations and in the hypersynchronous electrical activity of epilepsy. PMID:17640909

How Hedstrom files fail during clinical use? A retrieval study based on SEM, optical microscopy and micro-XCT analysis.

PubMed

Zinelis, Spiros; Al Jabbari, Youssef S

2018-05-01

This study was conducted to evaluate the failure mechanism of clinically failed Hedstrom (H)-files. Discarded H-files (n=160) from #8 to #40 ISO sizes were collected from different dental clinics. Retrieved files were classified according to their macroscopic appearance and they were investigated under scanning electron microscopy (SEM) and X-ray micro-computed tomography (mXCT). Then the files were embedded in resin along their longitudinal axis and after metallographic grinding and polishing, studied under an incident light microscope. The macroscopic evaluation showed that small ISO sizes (#08-#15) failed by extensive plastic deformation, while larger sizes (≥#20) tended to fracture. Light microscopy and mXCT results coincided showing that unused and plastically deformed files were free of internal defects, while fractured files demonstrate the presence of intense cracking in the flute region. SEM analysis revealed the presence of striations attributed to the fatigue mechanism. Secondary cracks were also identified by optical microscopy and their distribution was correlated to fatigue under bending loading. Experimental results demonstrated that while overloading of cutting instruments is the predominating failure mechanism of small file sizes (#08-#15), fatigue should be considered the fracture mechanism for larger sizes (≥#20).

Vaccinia Virus Mutations in the L4R Gene Encoding a Virion Structural Protein Produce Abnormal Mature Particles Lacking a Nucleocapsid

PubMed Central

Moussatche, Nissin; Condit, Richard C.

2014-01-01

ABSTRACT Electron micrographs from the 1960s revealed the presence of an S-shaped tubular structure in the center of the vaccinia virion core. Recently, we showed that packaging of virus transcription enzymes is necessary for the formation of the tubular structure, suggesting that the structure is equivalent to a nucleocapsid. Based on this study and on what is known about nucleocapsids of other viruses, we hypothesized that in addition to transcription enzymes, the tubular structure also contains the viral DNA and a structural protein as a scaffold. The vaccinia virion structural protein L4 stands out as the best candidate for the role of a nucleocapsid structural protein because it is abundant, it is localized in the center of the virion core, and it binds DNA. In order to gain more insight into the structure and relevance of the nucleocapsid, we analyzed thermosensitive and inducible mutants in the L4R gene. Using a cryo-fixation method for electron microscopy (high-pressure freezing followed by freeze-substitution) to preserve labile structures like the nucleocapsid, we were able to demonstrate that in the absence of functional L4, mature particles with defective internal structures are produced under nonpermissive conditions. These particles do not contain a nucleocapsid. In addition, the core wall of these virions is abnormal. This suggests that the nucleocapsid interacts with the core wall and that the nucleocapsid structure might be more complex than originally assumed. IMPORTANCE The vaccinia virus nucleocapsid has been neglected since the 1960s due to a lack of electron microscopy techniques to preserve this labile structure. With the advent of cryo-fixation techniques, like high-pressure freezing/freeze-substitution, we are now able to consistently preserve and visualize the nucleocapsid. Because vaccinia virus early transcription is coupled to the viral core structure, detailing the structure of the nucleocapsid is indispensable for determining the mechanisms of vaccinia virus core-directed transcription. The present study represents our second attempt to understand the structure and biological significance of the nucleocapsid. We demonstrate the importance of the protein L4 for the formation of the nucleocapsid and reveal in addition that the nucleocapsid and the core wall may be associated, suggesting a higher level of complexity of the nucleocapsid than predicted. In addition, we prove the utility of high-pressure freezing in preserving the vaccinia virus nucleocapsid. PMID:25253347

Nontraumatic tibial polyethylene insert cone fracture in mobile-bearing posterior-stabilized total knee arthroplasty.

PubMed

Tanikake, Yohei; Hayashi, Koji; Ogawa, Munehiro; Inagaki, Yusuke; Kawate, Kenji; Tomita, Tetsuya; Tanaka, Yasuhito

2016-12-01

A 72-year-old male patient underwent mobile-bearing posterior-stabilized total knee arthroplasty for osteoarthritis. He experienced a nontraumatic polyethylene tibial insert cone fracture 27 months after surgery. Scanning electron microscopy of the fracture surface of the tibial insert cone suggested progress of ductile breaking from the posterior toward the anterior of the cone due to repeated longitudinal bending stress, leading to fatigue breaking at the anterior side of the cone, followed by the tibial insert cone fracture at the anterior side of the cone, resulting in fracture at the base of the cone. This analysis shows the risk of tibial insert cone fracture due to longitudinal stress in mobile-bearing posterior-stabilized total knee arthroplasty in which an insert is designed to highly conform to the femoral component.

Subcritical crack growth and other time- and environment-dependent behavior in crustal rocks

NASA Technical Reports Server (NTRS)

Swanson, P. L.

1984-01-01

Stable crack growth strongly influences both the fracture strength of brittle rocks and some of the phenomena precursory to catastrophic failure. Quantification of the time and environment dependence of fracture propagation is attempted with the use of a fracture mechanics technique. Some of the difficulties encountered when applying techniques originally developed for simple synthetic materials to complex materials like rocks are examined. A picture of subcritical fracture propagation is developed that embraces the essential ingredients of the microstructure, a microcrack process zone, and the different roles that the environment plays. To do this, the results of (1) fracture mechanics experiments on five rock types, (2) optical and scanning electron microscopy, (3) studies of microstructural aspects of fracture in ceramics, and (4) exploratory tests examining the time-dependent response of rock to the application of water are examined.

Rehydration of freeze-dried and convective dried boletus edulis mushrooms: effect on some quality parameters.

PubMed

Hernando, I; Sanjuán, N; Pérez-Munuera, I; Mulet, A

2008-10-01

Quality of rehydrated products is a key aspect linked to rehydration conditions. To assess the effect of rehydration temperature on some quality parameters, experiments at 20 and 70 degrees C were performed with convective dried and freeze-dried Boletus edulis mushrooms. Rehydration characteristics (through Peleg's parameter, k(1), and equilibrium moisture, W(e)), texture (Kramer), and microstructure (Cryo-Scanning Electron Microscopy) were evaluated. Freeze-dried samples absorbed water more quickly and attained higher W(e) values than convective dried ones. Convective dehydrated samples rehydrated at 20 degrees C showed significantly lower textural values (11.9 +/- 3.3 N/g) than those rehydrated at 70 degrees C (15.7 +/- 1.2 N/g). For the freeze-dried Boletus edulis, the textural values also exhibited significant differences, being 8.2 +/- 1.3 and 10.5 +/- 2.3 N/g for 20 and 70 degrees C, respectively. Freeze-dried samples showed a porous structure that allows rehydration to take place mainly at the extracellular level. This explains the fact that, regardless of temperature, freeze-dried mushrooms absorbed water more quickly and reached higher W(e) values than convective dried ones. Whatever the dehydration technique used, rehydration at 70 degrees C produced a structural damage that hindered water absorption; consequently lower W(e) values and higher textural values were attained than when rehydrating at 20 degrees C.

Aldehyde dehydrogenase, Ald4p, is a major component of mitochondrial fluorescent inclusion bodies in the yeast Saccharomyces cerevisiae

PubMed Central

Misonou, Yoshiko; Kikuchi, Maiko; Sato, Hiroshi; Inai, Tomomi; Kuroiwa, Tsuneyoshi; Tanaka, Kenji; Miyakawa, Isamu

2014-01-01

ABSTRACT When Saccharomyces cerevisiae strain 3626 was cultured to the stationary phase in a medium that contained glucose, needle-like structures that emitted autofluorescence were observed in almost all cells by fluorescence microscopy under UV excitation. The needle-like structures completely overlapped with the profile of straight elongated mitochondria. Therefore, these structures were designated as mitochondrial fluorescent inclusion bodies (MFIBs). The MFIB-enriched mitochondrial fractions were successfully isolated and 2D-gel electrophoresis revealed that a protein of 54 kDa was only highly concentrated in the fractions. Determination of the N-terminal amino acid sequence of the 54-kDa protein identified it as a mitochondrial aldehyde dehydrogenase, Ald4p. Immunofluorescence microscopy showed that anti-Ald4p antibody specifically stained MFIBs. Freeze-substitution electron microscopy demonstrated that cells that retained MFIBs had electron-dense filamentous structures with a diameter of 10 nm in straight elongated mitochondria. Immunoelectron microscopy showed that Ald4p was localized to the electron-dense filamentous structures in mitochondria. These results together showed that a major component of MFIBs is Ald4p. In addition, we demonstrate that MFIBs are common features that appear in mitochondria of many species of yeast. PMID:24771619

The ability to survive intracellular freezing in nematodes is related to the pattern and distribution of ice formed.

PubMed

Raymond, Méliane R; Wharton, David A

2016-07-01

A few species of nematodes can survive extensive intracellular freezing throughout all their tissues, an event that is usually thought to be fatal to cells. How are they able to survive in this remarkable way? The pattern and distribution of ice formed, after freezing at -10°C, can be observed using freeze substitution and transmission electron microscopy, which preserves the former position of ice as white spaces. We compared the pattern and distribution of ice formed in a nematode that survives intracellular freezing well (Panagrolaimus sp. DAW1), one that survives poorly (Panagrellus redivivus) and one with intermediate levels of survival (Plectus murrayi). We also examined Panagrolaimus sp. in which the survival of freezing had been compromised by starvation. Levels of survival were as expected and the use of vital dyes indicated cellular damage in those that survived poorly (starved Panagrolaimus sp. and P. murrayi). In fed Panagrolaimus sp. the intracellular ice spaces were small and uniform, whereas in P. redivivus and starved Panagrolaimus sp. there were some large spaces that may be causing cellular damage. The pattern and distribution of ice formed was different in P. murrayi, with a greater number of individuals having no ice or only small intracellular ice spaces. Control of the size of the ice formed is thus important for the survival of intracellular freezing in nematodes. © 2016. Published by The Company of Biologists Ltd.

Fractography of the interlaminar fracture of carbon-fibre epoxy composites

NASA Technical Reports Server (NTRS)

Bascom, W. D.; Boll, D. J.; Fuller, B.; Phillips, P. J.

1985-01-01

The failed surfaces of interlaminar fracture (mode I) specimens of AS4/3501-6 were examined using scanning electron microscopy. The principal fracture features were fiber pull-out (bundles and single fibers), hackle markings, and regions of smooth resin fracture. Considerable (30 to 50 percent) relaxation of the deformed resin occurred when the specimens were heated above the matrix glass transition temperature. This relaxation was taken as evidence of extensive shear yielding of the resin during the fracture process. Some of the fractography features are discussed in terms of transverse tensile stresses and peeling stresses acting on the fibers. In some instances these localized stresses focus failure close to the resin-fiber interface, which can be mistakenly interpreted as interfacial failure and low fiber-resin adhesion.

Three dimensional structure of the distal condyles of the third metacarpal bone of the horse.

PubMed

Boyde, A; Haroon, Y; Jones, S J; Riggs, C M

1999-03-01

This study examined the three-dimensional (3D) microarchitecture of regions of the equine third metacarpal bone (McIII) commonly involved in distal condylar fractures. Limbs were obtained from Thoroughbred horses (neonates to age 24 years) destroyed for inoperable fractures and a variety of other conditions. Beams, blocks and sections were cut in the principal axes, some embedded in PMMA and others examined unembedded. Several methods were used to study the 3D structure, including conventional and confocal optical microscopy, scanning electron microscopy (SEM) and radiography. The mineralised articular cartilage tends to cleave in the sagittal plane. Proximal to the subchondral bone, the main trabeculae are robust plates running in the sagittal direction with less significant mediolateral connections. Small blood vessel canals lie inside the sagittal plates. This structure gives maximum strength and protection in the sagittal plane in which the bone rotates, but offers minimal resistance to fracture propagation in this plane. The anatomical course of the common distal condylar fractures of the third metacarpal bones can be explained by underlying anisotropic structural features of the mineralised tissues.

Response of resin transfer molded (RTM) composites under reversed cyclic loading

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mahfuz, H.; Haque, A.; Yu, D.

1996-01-01

Compressive behavior and the tension-compression fatigue response of resin transfer molded IM7 PW/PR 500 composite laminate with a circular notch have been studied. Fatigue damage characteristics have been investigated through the changes in the laminate strength and stiffness by gradually incrementing the fatigue cycles at a preselected load level. Progressive damage in the surface of the laminate during fatigue has been investigated using cellulose replicas. Failure mechanisms during static and cyclic tests have been identified and presented in detail. Extensive debonding of filaments and complete fiber bundle fracture accompanied by delamination were found to be responsible for fatigue failures, whilemore » fiber buckling, partial fiber fracture and delamination were characterized as the failure modes during static tests. Weibull analysis of the static, cyclic and residual tests have been performed and described in detail. Fractured as well as untested specimens were C-scanned, and the progressive damage growth during fatigue is presented. Optical Microscopy (OM) and Scanning Electron Microscopy (SEM) for the fractured specimen were also performed and the analysis of the failure behavior is presented.« less

Two Classes of Gap Junction Channels Mediate Soma-Germline Interactions Essential for Germline Proliferation and Gametogenesis in Caenorhabditis elegans

PubMed Central

Starich, Todd A.; Hall, David H.; Greenstein, David

2014-01-01

In all animals examined, somatic cells of the gonad control multiple biological processes essential for germline development. Gap junction channels, composed of connexins in vertebrates and innexins in invertebrates, permit direct intercellular communication between cells and frequently form between somatic gonadal cells and germ cells. Gap junctions comprise hexameric hemichannels in apposing cells that dock to form channels for the exchange of small molecules. Here we report essential roles for two classes of gap junction channels, composed of five innexin proteins, in supporting the proliferation of germline stem cells and gametogenesis in the nematode Caenorhabditis elegans. Transmission electron microscopy of freeze-fracture replicas and fluorescence microscopy show that gap junctions between somatic cells and germ cells are more extensive than previously appreciated and are found throughout the gonad. One class of gap junctions, composed of INX-8 and INX-9 in the soma and INX-14 and INX-21 in the germ line, is required for the proliferation and differentiation of germline stem cells. Genetic epistasis experiments establish a role for these gap junction channels in germline proliferation independent of the glp-1/Notch pathway. A second class of gap junctions, composed of somatic INX-8 and INX-9 and germline INX-14 and INX-22, is required for the negative regulation of oocyte meiotic maturation. Rescue of gap junction channel formation in the stem cell niche rescues germline proliferation and uncovers a later channel requirement for embryonic viability. This analysis reveals gap junctions as a central organizing feature of many soma–germline interactions in C. elegans. PMID:25195067

An exploratory investigation of cumulative shock fatigue.

NASA Technical Reports Server (NTRS)

Simonson, D.; Byrne, J. G.

1972-01-01

A simple device for producing cumulative shock loading in solids is described. The device uses a ballistic-impact-driven projectile to introduce high-stress waves into a solid. The impact time and load amplitude can be varied to produce fracture in one or several impacts in PMMA rods. The wavefront approached a square wave shape. Materials other than PMMA were loaded to failure to demonstrate the versatility of the device. Fracture morphologies observed with optical and scanning-electron microscopy are described.

Fracture surface analysis in composite and titanium bonding: Part 1: Titanium bonding

NASA Technical Reports Server (NTRS)

Sanderson, K. A.; Wightman, J. P.

1985-01-01

Fractured lap shear Ti 6-4 adherends bonded with polyphenyquinoxaline (PPQ) and polysulfone were analyzed. The effects of adherend pretreatment, stress level, thermal aging, anodizing voltage, and modified adhesive of Ti 6-4 adherend bonded with PPQ on lap shear strength were studied. The effect of adherend pretreatment on lap shear strength was investigated for PS samples. Results of scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) used to study the surface topography and surface composition are also discussed.

Influence of Emulsified Asphalt on the Mechanical Property and Microstructure of Cement-Stabilized Gravel under Freezing and Thawing Cycle Conditions.

PubMed

Wang, Yiqi; Tan, Yiqiu; Guo, Meng; Wang, Xinglong

2017-05-06

Properties of cement-stabilized gravel modified by emulsified asphalt under freezing and thawing cycle conditions were investigated by adjusting the dosage of cement. Mercury intrusion porosimetry (MIP) and Scanning electron microscopy (SEM) were introduced to analyze the influential mechanism. The results indicate that cement emulsified asphalt stabilized gravel with 5 wt % of cement performed well in both mechanics and frost-resistance. Although the addition of emulsified asphalt would lead to a partial decrease of strength, it can extend the process of strength loss and improve the freezing resistance. The main reason for this is that the permeability can be improved by the filling effects of emulsified asphalt. The frost-heave stress caused by the phase transition of water can also be remitted by emulsified asphalt, the elasticity modulus of which is much lower than the matrix. The generating speed of the micro crack can also be slowed down by emulsified asphalt.

Description and analysis of cored hydraulic fractures -- Lost Hills field, Kern County, California

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fast, R.E.; Murer, A.S.; Timmer, R.S.

1994-05-01

An inclined observation well was drilled in shallow (2,000 ft) Opal-A diatomite. Seven sand-propped hydraulic fractures were cored and recovered. The hydraulic fractures were found within 5[degree] of the azimuth measured with tilt meters and were tilted 15[degree] from vertical, oriented perpendicular to the formation bedding dip. Hydraulic fractures widths ranged from less than one sand grain (40/60 mesh) to 0.4 in. Scanning electron microscopy (SEM) examination of fracture faces showed no damage to the matrix from proppant embedment or compaction, and no evidence of guard residue was detected in the proppant pack or on the formation face. Fractures appearmore » to be considerably longer than modeled. Three closely spaced fractures are interpreted to be branches of a single hydraulic fracture treatment. This paper presents a description of the fractures recovered during coring in Well OO2. Findings related to fracture dimensions and orientations, fracture sources, fracture permeability measurements, and fracture characteristics (proppant embedment, presence of gel residue) are presented. Implications related to field development are discussed.« less

Supplementing zinc oxide nanoparticles to cryopreservation medium minimizes the freeze-thaw-induced damage to spermatozoa.

PubMed

Isaac, Ann V; Kumari, Sandhya; Nair, Ramya; Urs, Deepak Raj; Salian, Sujith Raj; Kalthur, Guruprasad; Adiga, Satish Kumar; Manikkath, Jyothsna; Mutalik, Srinivas; Sachdev, Divya; Pasricha, Renu

2017-12-16

The sperm DNA integrity post cryopreservation of human semen samples is one of the serious concerns in human infertility treatment. In the present study, the beneficial effects of zinc oxide nanoparticles in preserving the functional ability of spermatozoa was explored. Ejaculates of normozoospermic men cryopreserved along with Zinc oxide nanoparticles (ZnONPs) exhibited non-significantly higher percentage of total and progressive motility in frozen-thawed samples compared to control. The sperm chromatin damage and malondialdehyde (MDA) level was significantly lower in ZnONPs group (P < 0.01 and P < 0.05 respectively) and the spermatozoa's ability to undergo acrosome reaction was also unaltered. Fluorescence microscopy and High resolution transmission electron microscopy analysis demonstrated that the ZnONPs do not penetrate the membrane of spermatozoa but stay around the spermatozoa. In conclusion, the presence of ZnONPs during cryopreservation appears to be beneficial to the spermatozoa as they withstand freeze-thaw process competently better than control, without any adverse effect shown. Copyright © 2017 Elsevier Inc. All rights reserved.

Human oocyte cryopreservation and the fate of cortical granules.

PubMed

Ghetler, Yehudith; Skutelsky, Ehud; Ben Nun, Isaac; Ben Dor, Liah; Amihai, Dina; Shalgi, Ruth

2006-07-01

To examine the effect of the commonly used oocyte cryopreservation protocol on the cortical granules (CGs) of human immature germinal vesicle (GV) and mature metaphase II (MII) oocytes. Laboratory study. IVF unit. Unfertilized, intracytoplasmic sperm injected (ICSI) oocytes, and immature oocytes were cryopreserved using a slow freezing-rapid thawing program with 1,2-propanediol (PROH) as a cryoprotectant. Cortical granule exocytosis (CGE) was assessed by either confocal microscopy or transmission electron microscopy (TEM). The survival rates of frozen-thawed oocytes (mature and immature) were significantly lower compared with zygotes. Both mature and immature oocytes exhibited increased fluorescence after cryopreservation, indicating the occurrence of CGE. Mere exposure of oocytes to cryoprotectants induced CGE of 70% the value of control zygotes. The TEM revealed a drastic reduction in the amount of CGs at the cortex of frozen-thawed GV and MII oocytes, as well as appearance of vesicles in the ooplasm. The commonly used PROH freezing protocol for human oocytes resulted in extensive CGE. This finding explains why ICSI is needed to achieve fertilization of frozen-thawed human oocytes.

A high resolution electron microscopy investigation of curvature in carbon nanotubes

NASA Astrophysics Data System (ADS)

Weldon, D. N.; Blau, W. J.; Zandbergen, H. W.

1995-07-01

Evidence for heptagon inclusion in multi-walled carbon nanotubes was sought in arc-produced carbon deposits. Transmission electron microscopy revealed many curved nanotubes although their relative abundance was low. Close examination of the micrographs in the regions of expected heptagon inclusion shows that the curvature is accomplished by folding or fracture of the lattice planes. This observed phenomenon contradicts the theoretical modelling studies which predict stable structures with negative curvature accomplished by heptagon/pentagon pairs. A possible explanation for curvature in single-walled tubes is presented based on a molecular mechanics geometry optimisation study of spa inclusion in a graphite sheet.

Direct observation of the effects of cellulose synthesis inhibitors using live cell imaging of Cellulose Synthase (CESA) in Physcomitrella patens.

PubMed

Tran, Mai L; McCarthy, Thomas W; Sun, Hao; Wu, Shu-Zon; Norris, Joanna H; Bezanilla, Magdalena; Vidali, Luis; Anderson, Charles T; Roberts, Alison W

2018-01-15

Results from live cell imaging of fluorescently tagged Cellulose Synthase (CESA) proteins in Cellulose Synthesis Complexes (CSCs) have enhanced our understanding of cellulose biosynthesis, including the mechanisms of action of cellulose synthesis inhibitors. However, this method has been applied only in Arabidopsis thaliana and Brachypodium distachyon thus far. Results from freeze fracture electron microscopy of protonemal filaments of the moss Funaria hygrometrica indicate that a cellulose synthesis inhibitor, 2,6-dichlorobenzonitrile (DCB), fragments CSCs and clears them from the plasma membrane. This differs from Arabidopsis, in which DCB causes CSC accumulation in the plasma membrane and a different cellulose synthesis inhibitor, isoxaben, clears CSCs from the plasma membrane. In this study, live cell imaging of the moss Physcomitrella patens indicated that DCB and isoxaben have little effect on protonemal growth rates, and that only DCB causes tip rupture. Live cell imaging of mEGFP-PpCESA5 and mEGFP-PpCESA8 showed that DCB and isoxaben substantially reduced CSC movement, but had no measureable effect on CSC density in the plasma membrane. These results suggest that DCB and isoxaben have similar effects on CSC movement in P. patens and Arabidopsis, but have different effects on CSC intracellular trafficking, cell growth and cell integrity in these divergent plant lineages.

The Septate Junction Protein Caspr is Required for Structural Support and Retention of KCNQ4 at Calyceal Synapses of Vestibular Hair Cells

PubMed Central

Sousa, Aurea D.; Andrade, Leonardo R.; Salles, Felipe T.; Pillai, Anilkumar M.; Buttermore, Elizabeth; Bhat, Manzoor A.; Kachar, Bechara

2009-01-01

The afferent innervation contacting the type I hair cells of the vestibular sensory epithelia form distinct calyceal synapses. The apposed pre- and post-synaptic membranes at this large area of synaptic contact are kept at a remarkably regular distance. Here, we show by freeze-fracture electron microscopy that a patterned alignment of proteins at the calyceal membrane resembles a type of intercellular junction that is rare in vertebrates, the septate junction (SJ). We found that a core molecular component of SJs, Caspr, colocalizes with the K+ channel KCNQ4 at the post-synaptic membranes of these calyceal synapses. Immunolabeling and ultrastructural analyses of Caspr knockout mice reveal that, in the absence of Caspr, the separation between the membranes of the hair cells and the afferent neurons is conspicuously irregular and often increased by an order of magnitude. In these mutants, KCNQ4 fails to cluster at the post-synaptic membrane and appears diffused along the entire calyceal membrane. Our results indicate that a septate-like junction provides structural support to calyceal synaptic contact with the vestibular hair cell, and that Caspr is required for the recruitment or retention of KCNQ4 at these synapses. PMID:19279247

Characterization of a Biomimetic Mesophase Composed of Nonionic Surfactants and an Aqueous Solvent.

PubMed

Adrien, V; Rayan, G; Reffay, M; Porcar, L; Maldonado, A; Ducruix, A; Urbach, W; Taulier, N

2016-10-11

We have investigated the physical and biomimetic properties of a sponge (L 3 ) phase composed of pentaethylene glycol monododecyl ether (C 12 E 5 ), a nonionic surfactant, an aqueous solvent, and a cosurfactant. The following cosurfactants, commonly used for solubilizing membrane proteins, were incorporated: n-octyl-β-d-glucopyranoside (β-OG), n-dodecyl-β-d-maltopyranoside (DDM), 4-cyclohexyl-1-butyl-β-d-maltoside (CYMAL-4), and 5-cyclohexyl-1-pentyl-β-d-maltoside (CYMAL-5). Partial phase diagrams of these systems were created. The L 3 phase was characterized using crossed polarizers, diffusion of a fluorescent probe by fluorescence recovery after pattern photobleaching (FRAPP), and freeze fracture electron microscopy (FFEM). By varying the hydration of the phase, we were able to tune the distance between adjacent bilayers. The characteristic distance (d b ) of the phase was obtained from small angle scattering (SAXS/SANS) as well as from FFEM, which yielded complementary d b values. These d b values were neither affected by the nature of the cosurfactant nor by the addition of membrane proteins. These findings illustrate that a biomimetic surfactant sponge phase can be created in the presence of several common membrane protein-solubilizing detergents, thus making it a versatile medium for membrane protein studies.

Chimerization of lactoferricin and lactoferrampin peptides strongly potentiates the killing activity against Candida albicans.

PubMed

Bolscher, Jan; Nazmi, Kamran; van Marle, Jan; van 't Hof, Wim; Veerman, Enno

2012-06-01

Bovine lactoferrin harbors 2 antimicrobial sequences (LFcin and LFampin), situated in close proximity in the N1-domain. To mimic their semi parallel configuration we have synthesized a chimeric peptide (LFchimera) in which these sequences are linked in a head-to-head fashion to the α- and ε-amino group, respectively, of a single lysine. In line with previously described bactericidal effects, this peptide was also a stronger candidacidal agent than the antimicrobial peptides LFcin17-30 and LFampin265-284, or a combination of these 2. Conditions that strongly reduced the candidacidal activities of LFcin17-30 and LFampin265-284, such as high ionic strength and energy depletion, had little influence on the activity of LFchimera. Freeze-fracture electron microscopy showed that LFchimera severely affected the membrane morphology, resulting in disintegration of the membrane bilayer and in an efflux of small and high molecular weight molecules such as ATP and proteins. The differential effects displayed by the chimeric peptide and a mixture of its constituent peptides clearly demonstrate the synergistic effect of linking these peptides in a fashion that allows a similar spatial arrangement as in the parent protein, suggesting that in bovine lactoferrrin the corresponding fragments act in concert in its candidacidal activity.

Correlative Light-Electron Fractography of Interlaminar Fracture in a Carbon-Epoxy Composite.

PubMed

Hein, Luis Rogerio de O; Campos, Kamila A de

2015-12-01

This work evaluates the use of light microscopes (LMs) as a tool for interlaminar fracture of polymer composite investigation with the aid of correlative fractography. Correlative fractography consists of an association of the extended depth of focus (EDF) method, based on reflected LM, with scanning electron microscopy (SEM) to evaluate interlaminar fractures. The use of these combined techniques is exemplified here for the mode I fracture of carbon-epoxy plain-weave reinforced composite. The EDF-LM is a digital image-processing method that consists of the extraction of in-focus pixels for each x-y coordinate in an image from a stack of Z-ordered digital pictures from an LM, resulting in a fully focused picture and a height elevation map for each stack. SEM is the most used tool for the identification of fracture mechanisms in a qualitative approach, with the combined advantages of a large focus depth and fine lateral resolution. However, LMs, with EDF software, may bypass the restriction on focus depth and present enough lateral resolution at low magnification. Finally, correlative fractography can provide the general comprehension of fracture processes, with the benefits of the association of different resolution scales and contrast modes.

The effect of microstructure on the fracture toughness of titanium alloys

NASA Technical Reports Server (NTRS)

Vanstone, R. H.; Low, J. R., Jr.; Shannon, J. L., Jr.

1974-01-01

The microstructure of the alpha titanium alloy Ti-5Al-2.5Sn and the metastable beta titanium alloy Beta 3 was examined. The material was from normal and extra low interstitial grade plates which were either air-cooled or furnace-cooled from an annealing treatment. Beta 3 was studied in alpha-aged and omega-aged plates which were heat treated to similar strength levels. Tensile and plane strain fracture toughness tests were conducted at room temperature on the alpha-aged material. The microstructure and fracture mechanisms of alloys were studied using optical metallography, electron microscopy, microprobe analyses, and texture pole figures. Future experiments are described.

Ductile-to-Brittle transition in <111> hadfield steel single crystals

NASA Astrophysics Data System (ADS)

Astafurova, E. G.; Chumlyakov, Yu. I.

2010-10-01

The deformation mechanism and the character of fracture of <111> austenitic Hadfield steel single crystals are studied during tension in the temperature range 77-673 K by scanning and transmission electron microscopy. It is found that a change in the fracture mechanism from ductile to brittle fracture according to the fractography criterion takes place at a higher temperature than that determined from a change in the elongation to failure of the single crystals. The ductile-to-brittle transition in the Hadfield steel single crystals is shown to be related to a high level of deforming stresses induced by solid-solution hardening and to mechanical twinning.

7A projection map of the S-layer protein sbpA obtained with trehalose-embedded monolayer crystals.

PubMed

Norville, Julie E; Kelly, Deborah F; Knight, Thomas F; Belcher, Angela M; Walz, Thomas

2007-12-01

Two-dimensional crystallization on lipid monolayers is a versatile tool to obtain structural information of proteins by electron microscopy. An inherent problem with this approach is to prepare samples in a way that preserves the crystalline order of the protein array and produces specimens that are sufficiently flat for high-resolution data collection at high tilt angles. As a test specimen to optimize the preparation of lipid monolayer crystals for electron microscopy imaging, we used the S-layer protein sbpA, a protein with potential for designing arrays of both biological and inorganic materials with engineered properties for a variety of nanotechnology applications. Sugar embedding is currently considered the best method to prepare two-dimensional crystals of membrane proteins reconstituted into lipid bilayers. We found that using a loop to transfer lipid monolayer crystals to an electron microscopy grid followed by embedding in trehalose and quick-freezing in liquid ethane also yielded the highest resolution images for sbpA lipid monolayer crystals. Using images of specimens prepared in this way we could calculate a projection map of sbpA at 7A resolution, one of the highest resolution projection structures obtained with lipid monolayer crystals to date.

Wide-range high-resolution transmission electron microscopy reveals morphological and distributional changes of endomembrane compartments during log to stationary transition of growth phase in tobacco BY-2 cells.

PubMed

Toyooka, Kiminori; Sato, Mayuko; Kutsuna, Natsumaro; Higaki, Takumi; Sawaki, Fumie; Wakazaki, Mayumi; Goto, Yumi; Hasezawa, Seiichiro; Nagata, Noriko; Matsuoka, Ken

2014-09-01

Rapid growth of plant cells by cell division and expansion requires an endomembrane trafficking system. The endomembrane compartments, such as the Golgi stacks, endosome and vesicles, are important in the synthesis and trafficking of cell wall materials during cell elongation. However, changes in the morphology, distribution and number of these compartments during the different stages of cell proliferation and differentiation have not yet been clarified. In this study, we examined these changes at the ultrastructural level in tobacco Bright yellow 2 (BY-2) cells during the log and stationary phases of growth. We analyzed images of the BY-2 cells prepared by the high-pressure freezing/freeze substitution technique with the aid of an auto-acquisition transmission electron microscope system. We quantified the distribution of secretory and endosomal compartments in longitudinal sections of whole cells by using wide-range gigapixel-class images obtained by merging thousands of transmission electron micrographs. During the log phase, all Golgi stacks were composed of several thick cisternae. Approximately 20 vesicle clusters (VCs), including the trans-Golgi network and secretory vesicle cluster, were observed throughout the cell. In the stationary-phase cells, Golgi stacks were thin with small cisternae, and only a few VCs were observed. Nearly the same number of multivesicular body and small high-density vesicles were observed in both the stationary and log phases. Results from electron microscopy and live fluorescence imaging indicate that the morphology and distribution of secretory-related compartments dramatically change when cells transition from log to stationary phases of growth. © The Author 2014. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oup.com.

Graphene oxide foams and their excellent adsorption ability for acetone gas

DOE Office of Scientific and Technical Information (OSTI.GOV)

He, Yongqiang; School of Science, Tianjin University, Tianjin 300072; Zhang, Nana

2013-09-01

Graphical abstract: - Highlights: • GO and RGO foams were prepared using a simple and green method, unidirectional freeze-drying. • The porous structure of the foams can be adjusted by changing GO concentrations. • GO and RGO foams show good adsorption efficiency for acetone gas. - Abstract: Graphene oxide (GO) and reduced graphene oxide (RGO) foams were prepared using a unidirectional freeze-drying method. These porous carbon materials were characterized by thermal gravimetric analysis, differential scanning calorimetry, X-ray photoelectron spectroscopy and scanning electron microscopy. The adsorption behavior of the two kinds of foams for acetone was studied. The result showed thatmore » the saturated adsorption efficiency of the GO foams was over 100%, and was higher than that of RGO foams and other carbon materials.« less

Fracture and damage evolution of fluorinated polymers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Brown, E. N.; Rae, P.; Orler, E. B.

2004-01-01

Fluoropolymers are often semi-crystalline in nature, with their linear chains forming complicated phases near room temperature and ambient pressure. The most widely used fluorocarbon polymer for engineering applications is polytetrafluoroethylene (PTFE), due to its extremely low coefficient of friction, outstanding resistance to corrosion, and excellent electrical properties. The phase structure of PTFE is complex with four well-characterized crystalline phases (three observed at atmospheric pressure) and substantial molecular motion well below the melting point. The first-order transition at 19 C between phases II and IV is an unraveling in the helical conformation. Further rotational disordering and untwisting of the helices occursmore » above 30 C giving way to phase I. The mechanical behavior, including fracture and damage evolution, of PTFE depends on the chain and segment motions dictated by crystalline phase microstructure. The presence of three unique phases at ambient pressure near room temperature implies that failure during standard operating conditions may be strongly dependent on the phase. This paper presents a preliminary study of fracture and damage evolution in PTFE with the effects of temperature-induced phase on fracture mechanisms. The quasi-static fracture of PTFE in the atmospheric pressure regime, over a range of temperatures, was found to be strongly phase dependent: phase II exhibits brittle-fracture, phase IV displays ductile-fracture with crazing and some stable crack growth, and plastic flow dominates phase 1. The bulk failure properties are correlated to failure mechanisms through fractography of the fracture surfaces (optical microscopy and scanning electron microscopy (SEM)).« less

Dynamic Mechanical Properties and Fracture Surface Morphologies of Core-Shell Rubber (CSR) Toughened Epoxy at Liquid Nitrogen (Ln2) Temperatures

NASA Technical Reports Server (NTRS)

Wang, J.; Magee, D.; Schneider, J. A.

2009-01-01

The dynamic mechanical properties and fracture surface morphologies were evaluated for a commercial epoxy resin toughened with two types of core-shell rubber (CSR) toughening agents (Kane Ace(Registered TradeMark) MX130 and MX960). The impact resistance (R) was evaluated by the resulting breaking energy measured in Charpy impact tests conducted on an instrumented drop tower. The resulting fracture surface morphologies were examined using Scanning Electron Microscopy (SEM). Fractographic observations of the CSR toughened epoxy tested at ambient temperature, showed a fracture as characterized by slender dendrite textures with large voids. The increasing number of dendrites and decreasing size of scale-like texture with more CSR particles corresponded with increased R. As the temperature decreased to Liquid Nitrogen (LN 2), the fracture surfaces showed a fracture characterized by a rough, torn texture containing many river markings and deep furrows.

Postharvest Ultrasound-Assisted Freeze-Thaw Pretreatment Improves the Drying Efficiency, Physicochemical Properties, and Macamide Biosynthesis of Maca (Lepidium meyenii).

PubMed

Chen, Jin-Jin; Gong, Peng-Fei; Liu, Yi-Lan; Liu, Bo-Yan; Eggert, Dawn; Guo, Yuan-Heng; Zhao, Ming-Xia; Zhao, Qing-Sheng; Zhao, Bing

2018-04-01

A novel technique of ultrasound-assisted freeze-thaw pretreatment (UFP) was developed to improve the drying efficiency of maca and bioactive amide synthesis in maca. The optimal UFP conditions are ultrasonic processing 90 min at 30 °C with 6 freeze-thaw cycles. Samples with freeze-thaw pretreatment (FP), ultrasound pretreatment (UP), and UFP were prepared for further comparative study. A no pretreatment (NP) sample was included as a control. The results showed that UFP improved the drying efficiency of maca slices, showing the highest effective moisture diffusivity (1.75 × 10 -9 m 2 /s). This result was further supported by low-field nuclear magnetic resonance (LF-NMR) analysis and scanning electron microscopy (SEM). The rehydration capacity and protein content of maca slices were improved by UFP. More importantly, contents of bioactive macamides and their biosynthetic precursors were increased in 2.5- and 10-fold, respectively. In conclusion, UFP is an efficient technique to improve drying efficiency, physicochemical properties, and bioactive macamides of maca, which can be applied in the industrial manufacture of maca products. © 2018 Institute of Food Technologists®.

An environmentally friendly method for the fabrication of reduced graphene oxide foam with a super oil absorption capacity.

PubMed

He, Yongqiang; Liu, Yue; Wu, Tao; Ma, Junkui; Wang, Xingrui; Gong, Qiaojuan; Kong, Weina; Xing, Fubao; Liu, Yu; Gao, Jianping

2013-09-15

Three kinds of graphene oxide (GO) foams were fabricated using different freezing methods (unidirectional freezing drying (UDF), non-directional freezing drying, and air freezing drying), and the corresponding reduced graphene oxide (RGO) foams were prepared by their thermal reduction of those GO foams. These RGO foams were characterized by Fourier transform infrared spectroscopy, thermal gravimetric analysis, X-ray diffraction, X-ray photoelectron spectroscopy, and scanning electron microscopy. The absorption process and the factors that influence the absorption capacity were investigated. The RGO foams are hydrophobic and showed extremely high absorbing abilities for organic liquids. The absorption capacity of the RGO foams made by UDF was higher than 100 g g(-1) for all the oils tested (gasoline, diesel oil, pump oil, lubricating oil and olive oil) and had the highest value of about 122 g g(-1) for olive oil. The oil absorption capacity of the GO foams was lower than that of the RGO foams, but for olive oil, the absorption capacity was still high than 70 g g(-1), which is higher than that of most oil absorbents. Copyright © 2013 Elsevier B.V. All rights reserved.

Comparative study of ice nucleating efficiency of K-feldspar in immersion and deposition freezing modes

NASA Astrophysics Data System (ADS)

Hiron, T.; Hoffmann, N.; Peckhaus, A.; Kiselev, A. A.; Leisner, T.; Flossmann, A. I.

2016-12-01

One of the main challenges in understanding the evolution of Earth's climate resides in the understanding the role of ice nucleation on the development of tropospheric clouds as well as its initiation. K-feldspar is known to be a very active ice nucleating particle and this study focuses on the characterization of its activity in two heterogeneous nucleation modes, immersion and deposition freezing.We use a newly built humidity-controlled cold stage allowing the simultaneous observation of up to 2000 identical 0.6-nanoliter droplets containing suspension of mineral dust particles. The droplets are first cooled down to observe immersion freezing, the obtained ice crystals are then evaporated and finally, the residual particles are exposed to the water vapor supersaturated with respect to ice.The ice nucleation abilities for the individual residual particles are then compared for the different freezing modes and correlation between immersion ice nuclei and deposition ice nuclei is investigated.Based on the electron microscopy analysis of the residual particles, we discuss the possible relationship between the ice nucleation properties of feldspar and its microstructure. Finally, we discuss the atmospheric implications of our experimental results, using DESCAM, a 1.5D bin-resolved microphysics model.

Recent advances in high-pressure freezing: equipment- and specimen-loading methods.

PubMed

McDonald, Kent L; Morphew, Mary; Verkade, Paul; Müller-Reichert, Thomas

2007-01-01

This chapter is an update of material first published by McDonald in the first volume of this book. Here, we discuss the improvements in the technology and the methodology of high-pressure freezing (HPF) since that article was published. First, we cover the latest innovation in HPF, the Leica EM PACT2. This machine differs significantly from the BAL-TEC HPM 010 high-pressure freezer, which was the main subject of the former chapter. The EM PACT2 is a smaller, portable machine and has an optional attachment, the Rapid Transfer System (RTS). This RTS permits easy and reproducible loading of the sample and allows one to do correlative light and electron microscopy with high time resolution. We also place more emphasis in this article on the details of specimen loading for HPF, which is considered the most critical phase of the whole process. Detailed procedures are described for how to high-pressure freeze cells in suspension, cells attached to substrates, tissue samples, or whole organisms smaller than 300 microm, and tissues or organisms greater than 300 microm in size. We finish the article with a brief discussion of freeze substitution and recommend some sample protocols for this procedure.

Mechanisms for Ductile Rupture - FY16 ESC Progress Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boyce, Brad L.; Carroll, Jay D.; Noell, Phillip

2017-01-01

Ductile rupture in metals is generally a multi-step process of void nucleation, growth, and coalescence. Particle decohesion and particle fracture are generally invoked as the primary microstructural mechanisms for room-temperature void nucleation. However, because high-purity materials also fail by void nucleation and coalescence, other microstructural features must also act as sites for void nucleation. Early studies of void initiation in high-purity materials, which included post-mortem fracture surface characterization using scanning electron microscopy (SEM) and high-voltage electron microscopy (HVEM) and in-situ HVEM observations of fracture, established the presence of dislocation cell walls as void initiation sites in high-purity materials. Direct experimentalmore » evidence for this contention was obtained during in-situ HVEM tensile tests of Be single crystals. Voids between 0.2 and 1 μm long appeared suddenly along dislocation cell walls during tensile straining. However, subsequent attempts to replicate these results in other materials, particularly α -Fe single crystals, were unsuccessful because of the small size of the dislocation cells, and these remain the only published in-situ HVEM observations of void nucleation at dislocation cell walls in the absence of a growing macrocrack. Despite this challenge, other approaches to studying void nucleation in high-purity metals also indicate that dislocation cell walls are nucleation sites for voids.« less

Transmission electron microscopy in molecular structural biology: A historical survey.

PubMed

Harris, J Robin

2015-09-01

In this personal, historic account of macromolecular transmission electron microscopy (TEM), published data from the 1940s through to recent times is surveyed, within the context of the remarkable progress that has been achieved during this time period. The evolution of present day molecular structural biology is described in relation to the associated biological disciplines. The contribution of numerous electron microscope pioneers to the development of the subject is discussed. The principal techniques for TEM specimen preparation, thin sectioning, metal shadowing, negative staining and plunge-freezing (vitrification) of thin aqueous samples are described, with a selection of published images to emphasise the virtues of each method. The development of digital image analysis and 3D reconstruction is described in detail as applied to electron crystallography and reconstructions from helical structures, 2D membrane crystals as well as single particle 3D reconstruction of icosahedral viruses and macromolecules. The on-going development of new software, algorithms and approaches is highlighted before specific examples of the historical progress of the structural biology of proteins and viruses are presented. Copyright © 2014 Elsevier Inc. All rights reserved.

Refixation of Osteochondral Fractures by an Ultrasound-Activated Pin System – An Ovine In Vivo Examination Using CT and Scanning Electron Microscope

PubMed Central

H, Neumann; A.P, Schulz; S, Breer; A, Unger; B, Kienast

2015-01-01

Background: Osteochondral injuries, if not treated appropriately, often lead to severe osteoarthritis of the affected joint. Without refixation of the osteochondral fragment, human cartilage only repairs these defects imperfectly. All existing refixation systems for chondral defects have disadvantages, for instance bad MRI quality in the postoperative follow-up or low anchoring forces. To address the problem of reduced stability in resorbable implants, ultrasound-activated pins were developed. By ultrasound-activated melting of the tip of these implants a higher anchoring is assumed. Aim of the study was to investigate, if ultrasound-activated pins can provide a secure refixation of osteochondral fractures comparing to conventional screw and conventional, resorbable pin osteosynthesis. CT scans and scanning electron microscopy should proovegood refixation results with no further tissue damage by the melting of the ultrasound-activated pins in comparison to conventional osteosynthesis. Methods: Femoral osteochondral fragments in sheep were refixated with ultrasound-activated pins (SonicPin™), Ethipins® and screws (Asnis™). The quality of the refixated fragments was examined after three month of full weight bearing by CT scans and scanning electron microscopy of the cartilage surface. Results: The CT examination found almost no statistically significant difference in the quality of refixation between the three different implants used. Concerning the CT morphology, ultrasound-activated pins demonstrated at least the same quality in refixation of osteochondral fragments as conventional resorbable pins or screws. The scanning electron microscopy showed no major surface damage by the three implants, especially any postulated cartilage damage induced by the heat of the ultrasound-activated pin. The screws protruded above the cartilage surface, which may affect the opposingtibial surface. Conclusion: Using CT scans and scanning electron microscopy, the SonicPin™, the Ethipin® and screws were at least equivalent in refixation quality of osteochondral fragments. PMID:25674184

Facile Preparation of Carbon Microcapsules Containing Phase-Change Material with Enhanced Thermal Properties

PubMed Central

Tahan Latibari, Sara; Mehrali, Mohammad; Mehrali, Mehdi; Mahlia, Teuku Meurah Indra; Metselaar, Hendrik Simon Cornelis

2014-01-01

This study describes the hydrothermal synthesis of a novel carbon/palmitic acid (PA) microencapsulated phase change material (MEPCM). The field emission scanning electron microscopy (FESEM) and high resolution transmission electron microscopy (HRTEM) images confirm that spherical capsules of uniform size were formed with a mean diameter of 6.42 μm. The melting and freezing temperature were found to be slightly lower than those of pure PA with little undercooling. The composite retained 75% of the latent heat of pure PA. Thermal stability of the MEPCM was found to be better than that of pure PA. The thermal conductivity of MEPCM was increased by as much as 41% at 30°C. Due to its good thermal properties and chemical and mechanical stability, the carbon/PA MEPCM displays a good potential for thermal energy storage systems. PMID:25054179

Effects of joint configuration for the arc welding of cast Ti-6Al-4V alloy rods in argon.

PubMed

Taylor, J C; Hondrum, S O; Prasad, A; Brodersen, C A

1998-03-01

Titanium and its alloys are more commonly used in prosthodontics and welding has become the most common modality for their joining. Studies on the welding of titanium and its alloys have not quantified this value, though its importance has been suggested. This study compared the strength and properties of the joint achieved at various butt joint gaps by the arc-welding of cast Ti-6Al-4V alloy tensile bars in an argon atmosphere. Forty of 50 specimens were sectioned and welded at four gaps. All specimens underwent tensile testing to determine ultimate tensile strength and percentage elongation, then oxygen analysis and scanning electron microscopy. As no more than 3 samples in any group of 10 actually fractured in the weld itself, a secondary analysis that involved fracture location was initiated. There were no differences in ultimate tensile strength or percentage elongation between specimens with weld gaps of 0.25, 0.50, 0.75, and 1.00 mm and the as-cast specimens. There were no differences in ultimate tensile strength between specimens fracturing in the weld and those fracturing in the gauge in welded specimens; however, as-cast specimens demonstrated a higher ultimate tensile strength than welded specimens that fractured in the weld. Specimens that fractured in the weld site demonstrated less ductility than those that fractured in the gauge in both welded and as-cast specimens, as confirmed by scanning electron microscopy examination. The weld wire showed an oxygen scavenging effect from the as-cast parent alloy. The effects of the joint gap were not significant, whereas the characteristics of the joint itself were, which displayed slightly lower strength and significantly lower ductility (and thus decreased toughness). The arc-welding of cast titanium alloy in argon atmosphere appears to be a reliable and efficient prosthodontic laboratory modality producing predictable results, although titanium casting and joining procedures must be closely controlled to minimize heat effects and oxygen contamination.

Porosity, petrophysics and permeability of the Whitby Mudstone (UK)

NASA Astrophysics Data System (ADS)

Houben, M.; Barnhoorn, A.; Hardebol, N.; Ifada, M.; Boersma, Q.; Douma, L.; Peach, C. J.; Bertotti, G.; Drury, M. R.

2016-12-01

Typically pore diameters in shales range from the µm down to the nm scale and the effective permeability of shale reservoirs is a function of the interconnectivity between the pore space and the natural fracture network present. The length and spacing of mechanical induced and natural fractures is one of the factors controlling gas produtivity from unconventional reservoirs. Permeability of the Whitby Mudstone measured on 1 inch cores was linked to microstructure and combined with natural fracture spacing present in outcrops along the Yorkshire coast (UK) to get insight into possible fluid pathways from reservoir to well. We used a combination of different techniques to characterize the porosity (gas adsorption, Scanning Electron Microscopy), mineralogy (X-Ray Fluorescence, X-Ray Diffraction, Scanning Electron Microscopy) and permeability (pressure step decay) of the Whitby Mudstone. In addition, we mapped the natural fracture network as present in outcrops along the Yorkshire coast (UK) at the 10-2-101m scale. Mineralogically we are dealing with a rock that is high in clay content and has an average organic matter content of about 10%. Results show a low porosity (max. 7%) as well as low permeability for the Whitby Mudstone. The permeability, measured parallel to bedding, depends on the confining pressure and is 86 nanodarcy at 10 MPa effective confining pressure and decreases to 16 nanodarcy at 40 MPa effective confining pressure. At the scale of observation the average distance to nearest natural fracture is in the order of 0.13 meter and 90 percent of all matrix elements are spaced within 0.4 meter to the nearest fracture. By assuming darcy flow, a permeability of 100 nanodarcy and 10% of overpressure we calculated that for the Whitby mudstone most of the gas resides in the matrix for less than 60 days until it reaches the fracture network.

Structural organisation and lipid composition of the epicuticular accessory layer of infective larvae of Trichinella spiralis.

PubMed

Gounaris, K; Smith, V P; Selkirk, M E

1996-05-22

The epicuticle of infective larvae of Trichinella spiralis represents the interface between this intracellular nematode parasite and the cytosol of mammalian skeletal muscle. The macromolecular structures that make up the epicuticle were studied by freeze-fracture electron microscopy and compositional analysis. Three fracture planes were observed: one with a typical plasma membrane-type bilayer organisation which was overlaid by two extended layers of lipid in an inverted cylindrical configuration. This overall structure remained unchanged in response to variations in temperature between 20 degrees C and 45 degrees C. The lipid cylinders were on average 6.8 nm in diameter, with randomly-associated particles that were not dissociated by high-salt treatment, indicative of hydrophobically associated proteins. The majority of the lipids were non-polar, consisting of cholesterol, cholesterol esters, mono- and tri-glycerides, and free fatty acids. Three major classes of phospholipids were identified: phosphatidylethanolamine, phosphatidylglycerol and phosphatidylcholine. Total lipid extracts did not adopt an inverted cylindrical or micellar configuration on isolation, but formed flat sheets of lamellae as did the purified polar and non-polar fractions of the lipids. Isolated lipids did not undergo thermally-induced polymorphism between 20 degrees C and 60 degrees C and there was no pH dependency of the structures adopted. The fatty acid saturation levels of the phospholipids were compatible with the observation that they did not form polymorphic structures on isolation. We suggest that this unusual configuration is probably stabilised by the associated (glyco)proteins and may be required for selective permeation of nutrients from the host cell cytosol and/or for maintaining the high curvature of the parasite within the cell.

Comparison of three-dimensional printing and vacuum freeze-dried techniques for fabricating composite scaffolds

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Kai; Li, Ruixin; Jiang, Wenxue, E-mail: jiangortholivea@sina.cn

In this study, the performances of different preparation methods of the scaffolds were analyzed for chondrocyte tissue engineering. Silk fibroin/collagen (SF/C) was fabricated using a vacuum freeze-dried technique and by 3D printing. The porosity, water absorption expansion rates, mechanical properties, and pore sizes of the resulting materials were evaluated. The proliferation and metabolism of the cells was detected at different time points using an MTT assay. Cell morphologies and distributions were observed by histological analysis and scanning electron microscopy (SEM). The porosity, water absorption expansion rate, and Young’s modulus of the material obtained via 3D printing were significantly higher thanmore » those obtained by the freeze-dried method, while the pore size did not differ significantly between the two methods. MTT assay results showed that the metabolism of cells seeded on the 3D printed scaffolds was more viable than the metabolism on the freeze-dried material. H&E staining of the scaffolds revealed that the number of cells in the 3D printed scaffold was higher in comparison to a similar measurement on the freeze-dried material. Consequently, stem cells grew well inside the 3D printed scaffolds, as measured by SEM, while the internal structure of the freeze-dried scaffold was disordered. Compared with the freeze-dried technique, the 3D printed scaffold exhibited better overall performance and was more suitable for cartilage tissue engineering. - Highlights: • Silk fibroin/collagen was fabricated using 3D printing. • Physical characterization and Cell compatibility were compared. • 3D printed scaffold exhibited better overall performance.« less

Physicochemical characterization of commercial freeze-dried snake antivenoms.

PubMed

Herrera, María; Solano, Daniela; Gómez, Aarón; Villalta, Mauren; Vargas, Mariángela; Sánchez, Andrés; Gutiérrez, José María; León, Guillermo

2017-02-01

Freeze-drying is a process used to improve the stability of pharmaceutical proteins, including snake antivenoms. This additional step confers these with a higher stability in comparison to liquid formulations, especially in tropical regions where high temperatures could affect the activity of immunoglobulins. Currently, the knowledge about freeze-drying process conditions for snake antivenoms is very limited. Some of the scarce scientific works on this subject reported reconstitution times up to 90 min for these preparations, which could imply a delay in the beginning of the antivenom therapy at the clinical setting. Therefore there is a reasonable concern about whether freeze-dried antivenoms exhibit the desired attributes for solid pharmaceutical proteins. In this work, a physicochemical characterization of seven commercial freeze-dried snake antivenoms was performed based on tests recommended by the World Health Organization (WHO). No significant differences were observed between the products regarding macroscopic appearance of the solid cakes, reconstitution times, residual humidity and monomers content. On the other hand, total protein concentration, turbidity and electrophoretic profile were different among samples. Microscopic analysis by scanning electron microscopy showed no collapsed structure and, instead, most of the samples showed a characteristic protein morphology composed of smooth plates and channels. All the parameters tested in this study were according to literature recommendations and evidenced that, in spite of slight variations found for some products, formulation and freeze-drying conditions chosen by manufacturers are adequate to prevent aggregation and generate, in physicochemical terms, freeze-dried antivenoms of acceptable quality. Copyright © 2016 Elsevier Ltd. All rights reserved.

Characterizing the freezing behavior of liposomes as a tool to understand the cryopreservation procedures.

PubMed

Siow, Lee Fong; Rades, Thomas; Lim, Miang Hoong

2007-12-01

Freezing behaviors of egg yolk l-alpha-phosphatidylcholine (EPC) and 1,2-dipalmitoyl-rac-glycero-3-phosphocholine (DPPC) large unilamellar vesicles (LUV) were quantitatively characterized in relation to freezing temperatures, cooling rates, holding time, presence of sodium chloride and phospholipid phase transition temperature. Cooling of the EPC LUV showed an abrupt increase in leakage of the encapsulated carboxyfluorescein (CF) between -5 degrees C and -10 degrees C, which corresponded with the temperatures of the extraliposomal ice formation at around -7 degrees C. For the DPPC LUV, CF leakage started at -10 degrees C, close to the temperature of the extraliposomal ice formation; followed by a subsequent rapid increase in leakage between -10 degrees C and -25 degrees C. Scanning electron microscopy showed that both of these LUV were freeze-concentrated and aggregated at sub-freezing temperatures. We suggest that the formation of the extraliposomal ice and the decrease of the unfrozen fraction causes freeze-injury and leakage of the CF. The degree of leakage, however, differs between EPC LUV and DPPC LUV that inherently vary in their phospholipid phase transition temperatures. With increasing holding time, the EPC LUV were observed to have higher leakage when they were held at -15 degrees C compared to at -30 degrees C whilst leakage of the DPPC LUV was higher when holding at -40 degrees C than at -15 degrees C and -50 degrees C. At slow cooling rates, osmotic pressure across the bilayers may cause an additional stress to the EPC LUV. The present work elucidates freeze-injury mechanisms of the phospholipid bilayers through the liposomal model membranes.

Optimization of protectant, salinity and freezing condition for freeze-drying preservation of Edwardsiella tarda

NASA Astrophysics Data System (ADS)

Yu, Yongxiang; Zhang, Zheng; Wang, Yingeng; Liao, Meijie; Li, Bin; Xue, Liangyi

2017-10-01

Novel preservation condition without ultra-low temperature is needed for the study of pathogen in marine fishes. Freeze-drying is such a method usually used for preservation of terrigenous bacteria. However, studies using freeze-drying method to preserving marine microorganisms remain very limited. In this study, we optimized the composition of protectants during the freeze-drying of Edwardsiella tarda, a fish pathogen that causes systemic infection in marine fishes. We found that the optimal composition of protectant mixture contained trehalose (8.0%), skim milk (12.0%), sodium citrate (2.0%), serum (12.0%) and PVP (2.0%). Orthogonal and interaction analyses demonstrated the interaction between serum and skim milk or sodium citrate. The highest survival rate of E. tarda was observed when the concentration of NaCl was 10.0, 30.0 and between 5.0 and 10.0 g L-1 for preparing TSB medium, E. tarda suspension and protectant mixture, respectively. When E. tarda was frozen at -80°C or -40°C for 6 h, its survival rate was higher than that under other tested conditions. Under the optimized conditions, when the protectant mixture was used during freeze-drying process, the survival rate (79.63%-82.30%) of E. tarda was significantly higher than that obtained using single protectant. Scanning electron microscopy (SEM) image indicated that E. tarda was embedded in thick matrix with detectable aggregation. In sum, the protectant mixture may be used as a novel cryoprotective additive for E. tarda.

Endoplasmic Reticulum-Golgi Intermediate Compartment Membranes and Vimentin Filaments Participate in Vaccinia Virus Assembly

PubMed Central

Risco, Cristina; Rodríguez, Juan R.; López-Iglesias, Carmen; Carrascosa, José L.; Esteban, Mariano; Rodríguez, Dolores

2002-01-01

Vaccinia virus (VV) has a complex morphogenetic pathway whose first steps are poorly characterized. We have studied the early phase of VV assembly, when viral factories and spherical immature viruses (IVs) form in the cytoplasm of the infected cell. After freeze-substitution numerous cellular elements are detected around assembling viruses: membranes, ribosomes, microtubules, filaments, and unidentified structures. A double membrane is clearly resolved in the VV envelope for the first time, and freeze fracture reveals groups of tubules interacting laterally on the surface of the viroplasm foci. These data strongly support the hypothesis of a cellular tubulovesicular compartment, related to the endoplasmic reticulum-Golgi intermediate compartment (ERGIC), as the origin of the first VV envelope. Moreover, the cytoskeletal vimentin intermediate filaments are found around viral factories and inside the viroplasm foci, where vimentin and the VV core protein p39 colocalize in the areas where crescents protrude. Confocal microscopy showed that ERGIC elements and vimentin filaments concentrate in the viral factories. We propose that modified cellular ERGIC membranes and vimentin intermediate filaments act coordinately in the construction of viral factories and the first VV form through a unique mechanism of viral morphogenesis from cellular elements. PMID:11799179

Cyclic fatigue resistance of 3 different nickel-titanium reciprocating instruments in artificial canals.

PubMed

Higuera, Oscar; Plotino, Gianluca; Tocci, Luigi; Carrillo, Gabriela; Gambarini, Gianluca; Jaramillo, David E

2015-06-01

The purpose of this study was to evaluate the cyclic fatigue resistance of 3 different nickel-titanium reciprocating instruments. A total of 45 nickel-titanium instruments were tested and divided into 3 experimental groups (n = 15): group 1, WaveOne Primary instruments; group 2, Reciproc R25 instruments; and group 3, Twisted File (TF) Adaptive M-L1 instruments. The instruments were then subjected to cyclic fatigue test on a static model consisting of a metal block with a simulated canal with 60° angle of curvature and a 5-mm radius of curvature. WaveOne Primary, Reciproc R25, and TF Adaptive instruments were activated by using their proprietary movements, WaveOne ALL, Reciproc ALL, and TF Adaptive, respectively. All instruments were activated until fracture occurred, and the time to fracture was recorded visually for each file with a 1/100-second chronometer. Mean number of cycles to failure and standard deviations were calculated for each group, and data were statistically analyzed (P < .05). Instruments were also observed through scanning electron microscopy to evaluate type of fracture. Cyclic fatigue resistance of Reciproc R25 and TF Adaptive M-L1 was significantly higher than that of WaveOne Primary (P = .009 and P = .002, respectively). The results showed no statistically significant difference between TF Adaptive M-L1 and Reciproc R25 (P = .686). Analysis of the fractured portion under scanning electron microscopy indicated that all instruments showed morphologic characteristics of ductile fracture that were due to accumulation of metal fatigue. No statistically significant differences were found between the instruments tested except for WaveOne Primary, which showed the lowest resistance to cyclic fatigue. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Effect of Boron and Titanium Addition on the Hot Ductility of Low-Carbon Nb-Containing Steel

NASA Astrophysics Data System (ADS)

Liu, Wei-Jian; Li, Jing; Shi, Cheng-Bin; Huo, Xiang-Dong

2015-12-01

The effect of boron and titanium addition on the hot ductility of Nb-containing steel was investigated using hot tensile tests. The fracture surface and the quenched longitudinal microstructure were examined by optical microscopy (OM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The results showed that both steel samples had the similar change from 1,100°C to 700°C. The hot ductility of Nb-containing steel with boron and titanium addition was higher than the steel without boron and titanium in the temperature range of 900-750°C. Because the formation of intergranular ferrite was inhibited by solute boron segregating on the grain boundary, the formation of TiN changed the distribution of Nb- and boron-containing precipitates and improved the amount of intragranular ferrite.

Fracture behavior of silica nanoparticle filled epoxy resin

NASA Astrophysics Data System (ADS)

Dittanet, Peerapan

This dissertation involves the addition of silica nanoparticles to a lightly crosslinked, model epoxy resin and investigates the effect of nanosilica content and particle size on glass transition temperature (Tg), coefficient of thermal expansion (CTE), Young's modulus (E), yield stress, and fracture toughness. This study aims to understand the influence of silica nanoparticle size, bimodal particle size distribution and silica content on the toughening behavior. The toughening mechanisms were determined using scanning electron microscopy (SEM), transmission electron microscopy (TEM) and transmission optical microscopy (TOM). The approach identifies toughening mechanisms and develops a toughening model from unimodal-particle size systems first, then extends these concepts to various mixtures micron- and nanometer-size particles in a similar model epoxy. The experimental results revealed that the addition of nanosilica did not have a significant effect on Tg or the yield stress of epoxy resin, i.e. the yield stress and Tg remained constant regardless of nanosilica particle size. As expected, the addition of nanosilica had a significant impact on CTE, modulus and fracture toughness. The CTE values of nanosilica-filled epoxies were found to decrease with increasing nanosilica content, which can be attributed to the much lower CTE of the nanosilica fillers. Interestingly, the decreases in CTE showed strong particle size dependence. The Young's modulus was also found to significantly improve with addition of nanosilica and increase with increasing filler content. However, the particle size did not exhibit any effect on the Young's modulus. Finally, the fracture toughness and fracture energy showed significant improvements with the addition of nanosilica, and increased with increasing filler content. The effect of particle size on fracture toughness was negligible. Observation of the fracture surfaces using SEM and TOM showed evidence of debonding of nanosilica particles, matrix void growth, and matrix shear banding, which are credited for the increases in toughness for nanosilica-filled epoxy systems. Epoxy containing mixtures of two different size distributions of silica particles (42 micrometer and 23 nm-170nm particles) was explored for possible multiplicative toughening effect and to further understand the particle-epoxy interactions and toughening mechanisms of bimodal particle size distribution systems. The fracture toughness was improved by approximately 30% compared to that of the epoxy containing only one particle size of silica particles. The toughness improvement from the interaction of particle debonding from large particles and plastic void growth from small particles was clearly observed. The improvement in toughness occurred when the volume fraction ratio of the large and small particles was more than 50:50 ratios. The increased toughness was found to be additive not multiplicative effect.

An investigation of the loss of ductility in hydrogen charged beta-Ti alloys

NASA Technical Reports Server (NTRS)

Robertson, Ian M.

1995-01-01

The high strength, low density, and good corrosion resistance of Ti-based alloys make them candidate materials for a number of applications in the aerospace industry. A major limitation in the use of these alloys in the advanced hypersonic flight vehicle program is their susceptibility to hydrogen embrittlement. This study focuses on the hydrogen sensitivity of TIMETAL 21S beta-Ti alloy. The material received was in the form of grip-ends of failed tensile test samples which had been exposed to different charging conditions (combinations of hydrogen pressure and temperature). The samples received, the charging conditions, and their fracture mode are discussed. It can be seen that the fracture behavior changes from ductile to brittle with increasing hydrogen content, but the transition in behavior occurs for a small increase in hydrogen concentration. The aim of this program was to assess the microstructural differences between the ductile and brittle alloys to ascertain the embrittlement mechanism. A range of tools which included x-ray diffraction, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used.

[Ultrastructural organization of cytoplasmatic membrane of Anaerobacter polyendosporus studied by electron microscopic cryofractography].

PubMed

Duda, V I; Suzina, N E; Dmitriev, V V

2001-01-01

Anaerobacter polyendosporus cells do not have typical mesosomes. However, the analysis of this anaerobic multispore bacterium by electron microscopic cryofractography showed that its cytoplasmic membrane contains specific intramembrane structures in the form of flat lamellar inverted lipid membranes tenths of nanometers to several microns in size. It was found that these structures are located in the hydrophobic interior between the outer and inner leaflets of the cytoplasmic membrane and do not contain intramembrane particles that are commonly present on freeze-fracture replicas. The flat inverted lipid membranes were revealed in bacterial cells cultivated under normal growth conditions, indicating the existence of a complex-type compartmentalization in biological membranes, which manifests itself in the formation of intramembrane compartments having the appearance of vesicles and inverted lipid membranes.

Membrane damage during dilution, cooling and freezing-thawing of boar spermatozoa packaged in plastic bags.

PubMed

Ortman, K; Rodriguez-Martinez, H

1994-02-01

The objective of the present investigation was to determine the degree of membrane damage in boar spermatozoa during the course of a cryopreservation procedure, including thawing. Ejaculates from four fertile Swedish Yorkshire boars were frozen under controlled conditions in Teflon-plastic bags (2.5 ml) using 3% glycerol as cryoprotectant. Membrane integrity was monitored using supravital fluorescent dyes and confirmed by scanning electron microscopy. The results show that cooling to +5 degrees C significantly affected the permeability of the plasmalemma. Supercooling (-6 degrees C) during the freezing program did not further deteriorate the integrity of the spermatozoal membrane. The thawing procedure however, dramatically increased the frequency of spermatozoa with damaged plasma membranes. Thus particular attention must be paid on designing a better thawing procedure when dealing with boar semen frozen in plastic bags.

Effect of Local Crystallographic Texture on the Fissure Formation During Charpy Impact Testing of Low-Carbon Steel

NASA Astrophysics Data System (ADS)

Ghosh, Abhijit; Patra, Sudipta; Chatterjee, Arya; Chakrabarti, Debalay

2016-06-01

The severity of the formation of fissures (also known as splitting or delamination) on the fracture surface of Charpy impact-tested samples of a low-carbon steel has been found to increase with the decrease in finish rolling temperature [1093 K to 923 K (820 °C to 650 °C)]. Combined scanning electron microscopy and electron back-scattered diffraction study revealed that crystallographic texture was the prime factor responsible for the fissure formation. Through-thickness texture band composed of cube [Normal Direction (ND)║<001>] and gamma [ND║<111>] orientations developed during the inter-critical rolling treatment. Strain incompatibility between these two texture bands causes fissure cracking on the main fracture plane. A new approach based on the angle between {001} planes of neighboring crystals has been employed in order to estimate the `effective grain size,' which is used to determine the cleavage fracture stress on different planes of a sample. The severity of fissure formation was found to be directly related to the difference in cleavage fracture stress between the `main fracture plane' and `fissure plane.' Clustering of ferrite grains having cube texture promoted the fissure crack propagation along the transverse `fissure plane,' by increasing the `effective grain size' and decreasing the cleavage fracture stress on that plane.

Enhanced flexibility and electron-beam-controlled shape recovery in alumina-coated Au and Ag core-shell nanowires

NASA Astrophysics Data System (ADS)

Vlassov, Sergei; Polyakov, Boris; Vahtrus, Mikk; Mets, Magnus; Antsov, Mikk; Oras, Sven; Tarre, Aivar; Arroval, Tõnis; Lõhmus, Rünno; Aarik, Jaan

2017-12-01

The proper choice of coating materials and methods in core-shell nanowire (NW) engineering is crucial to assuring improved characteristics or even new functionalities of the resulting composite structures. In this paper, we have reported electron-beam-induced reversible elastic-to-plastic transition in Ag/Al2O3 and Au/Al2O3 NWs prepared by the coating of Ag and Au NWs with Al2O3 by low-temperature atomic layer deposition. The observed phenomenon enabled freezing the bent core-shell NW at any arbitrary curvature below the yield strength of the materials and later restoring its initially straight profile by irradiating the NW with electrons. In addition, we demonstrated that the coating efficiently protects the core material from fracture and plastic yield, allowing it to withstand significantly higher deformations and stresses in comparison to uncoated NW.

Connexin36 vs. connexin32, "miniature" neuronal gap junctions, and limited electrotonic coupling in rodent suprachiasmatic nucleus.

PubMed

Rash, J E; Olson, C O; Pouliot, W A; Davidson, K G V; Yasumura, T; Furman, C S; Royer, S; Kamasawa, N; Nagy, J I; Dudek, F E

2007-10-26

Suprachiasmatic nucleus (SCN) neurons generate circadian rhythms, and these neurons normally exhibit loosely-synchronized action potentials. Although electrotonic coupling has long been proposed to mediate this neuronal synchrony, ultrastructural studies have failed to detect gap junctions between SCN neurons. Nevertheless, it has been proposed that neuronal gap junctions exist in the SCN; that they consist of connexin32 or, alternatively, connexin36; and that connexin36 knockout eliminates neuronal coupling between SCN neurons and disrupts circadian rhythms. We used confocal immunofluorescence microscopy and freeze-fracture replica immunogold labeling to examine the distributions of connexin30, connexin32, connexin36, and connexin43 in rat and mouse SCN and used whole-cell recordings to re-assess electrotonic and tracer coupling. Connexin32-immunofluorescent puncta were essentially absent in SCN but connexin36 was relatively abundant. Fifteen neuronal gap junctions were identified ultrastructurally, all of which contained connexin36 but not connexin32, whereas nearby oligodendrocyte gap junctions contained connexin32. In adult SCN, one neuronal gap junction was >600 connexons, whereas 75% were smaller than 50 connexons, which may be below the limit of detectability by fluorescence microscopy and thin-section electron microscopy. Whole-cell recordings in hypothalamic slices revealed tracer coupling with neurobiotin in <5% of SCN neurons, and paired recordings (>40 pairs) did not reveal obvious electrotonic coupling or synchronized action potentials, consistent with few neurons possessing large gap junctions. However, most neurons had partial spikes or spikelets (often <1 mV), which remained after QX-314 [N-(2,6-dimethylphenylcarbamoylmethyl)triethylammonium bromide] had blocked sodium-mediated action potentials within the recorded neuron, consistent with spikelet transmission via small gap junctions. Thus, a few "miniature" gap junctions on most SCN neurons appear to mediate weak electrotonic coupling between limited numbers of neuron pairs, thus accounting for frequent detection of partial spikes and hypothetically providing the basis for "loose" electrical or metabolic synchronization of electrical activity commonly observed in SCN neuronal populations during circadian rhythms.

Experimental simulation of frost wedging-induced crack propagation in alpine rockwall

NASA Astrophysics Data System (ADS)

Jia, Hailiang; Leith, Kerry; Krautblatter, Michael

2016-04-01

Frost wedging is widely presumed to be the principal mechanism responsible for shattering jointed low-porosity rocks in high alpine rockwalls. The interaction of ice and rock physics regulates the efficacy of frost wedging. In order to better understand temporal aspects of this interaction, we present results of a series of laboratory experiments monitoring crack widening as a result of ice formation in an artificial crack (4mm wide, 80mm deep) cut 20 mm from the end of a rectangular granite block. Our results indicate that i) freezing direction plays a key role in determining the magnitude of crack widening; in short-term (1 day) experiments, maximum crack widening during top-down freezing (associated with 'autumn' conditions) was around 0.11mm, while inside-out freezing (resulting from 'spring' conditions) produced only 0.02 mm of deformation; ii) neither ice, nor water pressure (direct tension and hydraulic fracturing respectively) caused measurable irreversible crack widening during short-term tests, as the calculated maximum stress intensity at the crack tip was less than the fracture toughness of our granite sample; iii) development of ice pressure is closely related to the mechanical properties of the fracture in which it forms, and as such, the interaction of ice and rock is intrinsically dynamic; iv) irreversible crack widening (about 0.03mm) was only observed following a long-term (53 day) experiment representing a simplified transition from autumn to winter conditions. We suggest this is the result of stress corrosion aided by strong opening during freezing, and to a lesser degree by ice segregation up to one week after the initial freezing period, and downward migration of liquid water during the remainder of the test. Our results suggest the fundamental assumption of frost wedging, that rapid freezing from open ends of cracks can seal water inside the crack and thus cause damage through excessive stresses induced by volumetric expansion seems questionable.

Challenges in the Structure Determination of Self-Assembled Metallacages: What Do Cage Cavities Contain, Internal Vapor Bubbles or Solvent and/or Counterions?

PubMed

Givelet, Cecile C; Dron, Paul I; Wen, Jin; Magnera, Thomas F; Zamadar, Matibur; Čépe, Klára; Fujiwara, Hiroki; Shi, Yue; Tuchband, Michael R; Clark, Noel; Zbořil, Radek; Michl, Josef

2016-05-25

Proving the structures of charged metallacages obtained by metal ion coordination-driven solution self-assembly is challenging, and the common use of routine NMR spectroscopy and mass spectrometry is unreliable. Carefully determined diffusion coefficients from diffusion-ordered proton magnetic resonance (DOSY NMR) for six cages of widely differing sizes lead us to propose a structural reassignment of two molecular cages from a previously favored trimer to a pentamer or hexamer, and another from a trimer to a much higher oligomer, possibly an intriguing tetradecamer. In the former case, strong support for the reassignment to a larger cage is provided by an observation of a slow reversible transformation of the initially formed cage into a smaller but spectrally very similar one upon dilution. In the latter case, freeze-fracture transmission electron micrographs demonstrate that at least some of the solutions are colloidal, and high-resolution electron transmission and atomic force microscopy images are compatible with a tetradecamer but not a trimer. Comparison of solute partial molar volumes deduced from measurement of solution density with volumes anticipated from molecular models argues strongly against the presence of large voids (solvent vapor bubbles) in cages dissolved in nitromethane. The presence of bubbles was previously proposed in an attempt to account for the bilinear nature of the Eyring plot of the rate constant for pyridine ligand edge exchange reaction in one of the cages and for the unusual activation parameters in the high-temperature regime. An alternative interpretation is proposed now.

Processing, thermal and mechanical behaviour of PEI/MWCNT/carbon fiber nanostructured laminate

NASA Astrophysics Data System (ADS)

Santos, L. F. P.; Ribeiro, B.; Hein, L. R. O.; Botelho, E. C.; Costa, M. L.

2017-11-01

In this work, nanostructured composites of polyetherimide (PEI) with addition of functionalized multiwall carbon nanotube (MWCNT) were processed via solution mixing. After processing, these nanocomposites were evaluated by thermogravimetry (TGA), dynamic-mechanical analysis (DMA), scanning electron microscopy (SEM) and atomic force microscopy (AFM). Subsequently, the nanocomposite was processed with carbon fibers by using hot compression molding. In order to evaluate interlaminar fracture strength, the processed laminates were mechanically evaluated by interlaminar shear strength (ILSS) and compression shear test (CST). Also, the Weibull distribution was employed to help in the statistical treatment of the data obtained from the mechanical tests. With regards to the fracture of the specimens, optical microscopy was used for the evaluation of the material. The addition of 1 wt% of MWCNT in the polymer matrix increased both thermal stability and viscoelastic behavior of the material. These improvements positively impacted the mechanical properties, generating a 16% and 58% increase in the short-beam strength and apparent interlaminar shear, respectively. In addition, it can be verified from morphological analysis of the fracture a change in the failure mode of the laminate by the incorporation of MWCNT. This behavior can be proven from CST test where there was no presence of the shear force by compression.

Phosphatidylinositol 3,5-Bisphosphate-Rich Membrane Domains in Endosomes and Lysosomes.

PubMed

Takatori, Sho; Tatematsu, Tsuyako; Cheng, Jinglei; Matsumoto, Jun; Akano, Takuya; Fujimoto, Toyoshi

2016-02-01

Phosphatidylinositol 3,5-bisphosphate (PtdIns(3,5)P2 ) has critical functions in endosomes and lysosomes. We developed a method to define nanoscale distribution of PtdIns(3,5)P2 using freeze-fracture electron microscopy. GST-ATG18-4×FLAG was used to label PtdIns(3,5)P2 and its binding to phosphatidylinositol 3-phosphate (PtdIns(3)P) was blocked by an excess of the p40(phox) PX domain. In yeast exposed to hyperosmotic stress, PtdIns(3,5)P2 was concentrated in intramembrane particle (IMP)-deficient domains in the vacuolar membrane, which made close contact with adjacent membranes. The IMP-deficient domain was also enriched with PtdIns(3)P, but was deficient in Vph1p, a liquid-disordered domain marker. In yeast lacking either PtdIns(3,5)P2 or its effector, Atg18p, the IMP-deficient, PtdIns(3)P-rich membranes were folded tightly to make abnormal tubular structures, thus showing where the vacuolar fragmentation process is arrested when PtdIns(3,5)P2 metabolism is defective. In HeLa cells, PtdIns(3,5)P2 was significantly enriched in the vesicular domain of RAB5- and RAB7-positive endosome/lysosomes of the tubulo-vesicular morphology. This biased distribution of PtdIns(3,5)P2 was also observed using fluorescence microscopy, which further showed enrichment of a retromer component, VPS35, in the tubular domain. This is the first report to show segregation of PtdIns(3,5)P2 -rich and -deficient domains in endosome/lysosomes, which should be important for endosome/lysosome functionality. © 2015 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Two classes of gap junction channels mediate soma-germline interactions essential for germline proliferation and gametogenesis in Caenorhabditis elegans.

PubMed

Starich, Todd A; Hall, David H; Greenstein, David

2014-11-01

In all animals examined, somatic cells of the gonad control multiple biological processes essential for germline development. Gap junction channels, composed of connexins in vertebrates and innexins in invertebrates, permit direct intercellular communication between cells and frequently form between somatic gonadal cells and germ cells. Gap junctions comprise hexameric hemichannels in apposing cells that dock to form channels for the exchange of small molecules. Here we report essential roles for two classes of gap junction channels, composed of five innexin proteins, in supporting the proliferation of germline stem cells and gametogenesis in the nematode Caenorhabditis elegans. Transmission electron microscopy of freeze-fracture replicas and fluorescence microscopy show that gap junctions between somatic cells and germ cells are more extensive than previously appreciated and are found throughout the gonad. One class of gap junctions, composed of INX-8 and INX-9 in the soma and INX-14 and INX-21 in the germ line, is required for the proliferation and differentiation of germline stem cells. Genetic epistasis experiments establish a role for these gap junction channels in germline proliferation independent of the glp-1/Notch pathway. A second class of gap junctions, composed of somatic INX-8 and INX-9 and germline INX-14 and INX-22, is required for the negative regulation of oocyte meiotic maturation. Rescue of gap junction channel formation in the stem cell niche rescues germline proliferation and uncovers a later channel requirement for embryonic viability. This analysis reveals gap junctions as a central organizing feature of many soma-germline interactions in C. elegans. Copyright © 2014 by the Genetics Society of America.

Plastoglobules Are Lipoprotein Subcompartments of the Chloroplast That Are Permanently Coupled to Thylakoid Membranes and Contain Biosynthetic Enzymes

PubMed Central

Austin, Jotham R.; Frost, Elizabeth; Vidi, Pierre-Alexandre; Kessler, Felix; Staehelin, L. Andrew

2006-01-01

Plastoglobules are lipoprotein particles inside chloroplasts. Their numbers have been shown to increase during the upregulation of plastid lipid metabolism in response to oxidative stress and during senescence. In this study, we used state-of-the-art high-pressure freezing/freeze-substitution methods combined with electron tomography as well as freeze-etch electron microscopy to characterize the structure and spatial relationship of plastoglobules to thylakoid membranes in developing, mature, and senescing chloroplasts. We demonstrate that plastoglobules are attached to thylakoids through a half-lipid bilayer that surrounds the globule contents and is continuous with the stroma-side leaflet of the thylakoid membrane. During oxidative stress and senescence, plastoglobules form linkage groups that are attached to each other and remain continuous with the thylakoid membrane by extensions of the half-lipid bilayer. Using three-dimensional tomography combined with immunolabeling techniques, we show that the plastoglobules contain the enzyme tocopherol cyclase (VTE1) and that this enzyme extends across the surface monolayer into the interior of the plastoglobules. These findings demonstrate that plastoglobules function as both lipid biosynthesis and storage subcompartments of thylakoid membranes. The permanent structural coupling between plastoglobules and thylakoid membranes suggests that the lipid molecules contained in the plastoglobule cores (carotenoids, plastoquinone, and tocopherol [vitamin E]) are in a dynamic equilibrium with those located in the thylakoid membranes. PMID:16731586

Subcellular pigment distribution is altered under far-red light acclimation in cyanobacteria that contain chlorophyll f.

PubMed

Majumder, Erica L-W; Wolf, Benjamin M; Liu, Haijun; Berg, R Howard; Timlin, Jerilyn A; Chen, Min; Blankenship, Robert E

2017-11-01

Far-Red Light (FRL) acclimation is a process that has been observed in cyanobacteria and algae that can grow solely on light above 700 nm. The acclimation to FRL results in rearrangement and synthesis of new pigments and pigment-protein complexes. In this study, cyanobacteria containing chlorophyll f, Synechococcus sp. PCC 7335 and Halomicronema hongdechloris, were imaged as live cells with confocal microscopy. H. hongdechloris was further studied with hyperspectral confocal fluorescence microscopy (HCFM) and freeze-substituted thin-section transmission electron microscopy (TEM). Under FRL, phycocyanin-containing complexes and chlorophyll-containing complexes were determined to be physically separated and the synthesis of red-form phycobilisome and Chl f was increased. The timing of these responses was observed. The heterogeneity and eco-physiological response of the cells was noted. Additionally, a gliding motility for H. hongdechloris is reported.

Nature and morphology of fumed oxides and features of interfacial phenomena

NASA Astrophysics Data System (ADS)

Gun'ko, V. M.; Zarko, V. I.; Goncharuk, O. V.; Matkovsky, A. K.; Remez, O. S.; Skubiszewska-Zięba, J.; Wojcik, G.; Walusiak, B.; Blitz, J. P.

2016-03-01

Individual and complex fumed nanooxides were studied using high-resolution transmission electron microscopy, X-ray diffraction, ultraviolet-visible (UV-vis) spectroscopy, differential scanning calorimetry, nuclear magnetic resonance spectroscopy, adsorption, desorption (evaporation), and quantum chemical methods. For mixed nanooxides in contrast to simple and small nanoparticles of individual silica or titania, complex core-shell nanoparticles (50-200 nm in size) with titania or alumina cores and silica or alumina shells can be destroyed under high-pressure cryogelation (HPCG), mechnochemical activation (MCA) that also affect the structure of aggregates of nanoparticles and agglomerates of aggregates becoming more compacted. This is accompanied by changes in color from white to beige of different tints and changes in the UV-vis spectra in the 300-600 nm range, as well as changes in crystalline structure of alumina. Any treatment of 'soft' nanooxides affects the interfacial behavior of polar and nonpolar adsorbates. For some of them, the hysteresis loops become strongly open. Rearrangement of secondary particles affects the freezing-melting point depression. Clusterization of adsorbates bound in pores causes diminution of heat effects during phase transition (freezing, fusion). Freezing point depression and increasing melting point cause significant hysteresis freezing-melting effects for adsorbates bound to oxide nanoparticles. The study shows that complex nanooxides can be more sensitive to external actions than simple nanooxides such as silica.

Mass Determination of Rous Sarcoma Virus Virions by Scanning Transmission Electron Microscopy

PubMed Central

Vogt, Volker M.; Simon, Martha N.

1999-01-01

The internal structural protein of retroviruses, Gag, comprises most of the mass of the virion, and Gag itself can give rise to virus-like particles when expressed in appropriate cells. Previously the stoichiometry of Gag in virions was inferred from indirect measurements carried out 2 decades ago. We now have directly determined the masses of individual particles of the prototypic avian retrovirus, Rous sarcoma virus (RSV), by using scanning transmission electron microscopy. In this technique, the number of scattered electrons in the dark-field image integrated over an individual freeze-dried virus particle on a grid is directly proportional to its mass. The RSV virions had a mean mass of 2.5 × 108 Da, corresponding to about 1,500 Gag molecules per virion. The population of virions was not homogeneous, with about one-third to two-thirds of the virions deviating from the mean by more than 10% of the mass in two respective preparations. The mean masses for virions carrying genomes of 7.4 or 9.3 kb were indistinguishable, suggesting that mass variability is not due to differences in RNA incorporation. PMID:10400808

Fractography of human intact long bone by bending.

PubMed

Kimura, T; Ogawa, K; Kamiya, M

1977-05-27

Human intact tibiae were tested using the static bending method to learn about the relationship between the fracture surface and the failure mode. The bending test was applied to test pieces and to whole bones. The fracture surface was observed by scanning electron microscopy. The bone fracture is closely related to the architecture of the bone substance, especially to the direction of the Haversian canals and the lamellae. The failure mode and the sequence of the break line of the bone can be found out by the observation on the fracture surface. Hardly any crushing effects caused by the compressive force is seen. The mechanical properties of the fractured bone can be estimated to some extend by considering the direction of the break line and the failure mode. The strength calculated by the simple beam formula for elastic materials can not be obtained directly because of the plastic deformation of the bone. The results of the tensile test may be applied to the fracture using the static bending moment.

Studies of the structure of insulin fibrils by Fourier transform infrared (FTIR) spectroscopy and electron microscopy.

PubMed

Nielsen, L; Frokjaer, S; Carpenter, J F; Brange, J

2001-01-01

Fibril formation (aggregation) of insulin was investigated in acid media by visual inspection, transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. Insulin fibrillated faster in hydrochloric acid than in acetic acid at elevated temperatures, whereas the fibrillation tendencies were reversed at ambient temperatures. Electron micrographs showed that bovine insulin fibrils consisted of long fibers with a diameter of 5 to 10 nm and lengths of several microns. The fibrils appeared either as helical filaments (in hydrochloric acid) or arranged laterally in bundles (in acetic acid, NaCl). Freeze-thawing cycles broke the fibrils into shorter segments. FTIR spectroscopy showed that the native secondary structure of insulin was identical in hydrochloric acid and acetic acid, whereas the secondary structure of fibrils formed in hydrochloric acid was different from that formed in acetic acid. Fibrils of bovine insulin prepared by heating or agitating an acid solution of insulin showed an increased content of beta-sheet (mostly intermolecular) and a decrease in the intensity of the alpha-helix band. In hydrochloric acid, the frequencies of the beta-sheet bands depended on whether the fibrillation was induced by heating or agitation. This difference was not seen in acetic acid. Freeze-thawing cycles of the fibrils in hydrochloric acid caused an increase in the intensity of the band at 1635 cm(-1) concomitant with reduction of the band at 1622 cm(-1). The results showed that the structure of insulin fibrils is highly dependent on the composition of the acid media and on the treatment. Copyright 2001 Wiley-Liss Inc. and the American Pharmaceutical Association J Pharm Sci 90: 29-37, 2001

Influence of the geometry of curved artificial canals on the fracture of rotary nickel-titanium instruments subjected to cyclic fatigue tests.

PubMed

Lopes, Hélio P; Vieira, Márcia V B; Elias, Carlos N; Gonçalves, Lucio S; Siqueira, José F; Moreira, Edson J L; Vieira, Victor T L; Souza, Letícia C

2013-05-01

This study evaluated the influence of different features of canal curvature geometry on the number of cycles to fracture of a rotary nickel-titanium endodontic instrument subjected to a cyclic fatigue test. BioRaCe BR4C instruments (FKG Dentaire, La Chaux-de Fonds, Switzerland) were tested in 4 grooves simulating curved metallic artificial canals, each one measuring 1.5 mm in width, 20 mm in total length, and 3.5 mm in depth with a U-shaped bottom. The parameters of curvature including the radius and arc lengths and the position of the arc differed in the 4 canal designs. Fractured surfaces and helical shafts of the separated instruments were analyzed by scanning electron microscopy. The Student's t test showed that a significantly lower number of cycles to fracture values were observed for instruments tested in canals with the smallest radius, the longest arc, and the arc located in the middle portion of the canal. Scanning electron microscopic analysis of the fracture surfaces revealed morphologic characteristics of ductile fracture. Plastic deformation was not observed in the helical shaft of the fractured instruments. Curvature geometry including the radius and arc lengths and the position of the arc along the root canal influence the number of cycles to fracture of rotary nickel-titanium instruments subjected to flexural load. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Tensile Fracture Behavior and Failure Mechanism of Additively-Manufactured AISI 4140 Low Alloy Steel by Laser Engineered Net Shaping

PubMed Central

Kim, Hoyeol; Liu, Zhichao; Cong, Weilong; Zhang, Hong-Chao

2017-01-01

AISI 4140 powder was directly deposited on AISI 4140 wrought substrate using laser engineered net shaping (LENS) to investigate the compatibility of a LENS-deposited part with the substrate. Tensile testing at room temperature was performed to evaluate the interface bond performance and fracture behavior of the test specimens. All the samples failed within the as-deposited zone, indicating that the interfacial bond is stronger than the interlayer bond inside the deposit. The fracture surfaces were analyzed using scanning electron microscopy (SEM) and energy disperse X-ray spectrometry (EDS). Results show that the tensile fracture failure of the as-deposited part is primarily affected by lack-of-fusion defects, carbide precipitation, and oxide particles inclusions, which causes premature failure of the deposit by deteriorating the mechanical properties and structural integrity. PMID:29120374

Tensile Fracture Behavior and Failure Mechanism of Additively-Manufactured AISI 4140 Low Alloy Steel by Laser Engineered Net Shaping.

PubMed

Kim, Hoyeol; Liu, Zhichao; Cong, Weilong; Zhang, Hong-Chao

2017-11-09

AISI 4140 powder was directly deposited on AISI 4140 wrought substrate using laser engineered net shaping (LENS) to investigate the compatibility of a LENS-deposited part with the substrate. Tensile testing at room temperature was performed to evaluate the interface bond performance and fracture behavior of the test specimens. All the samples failed within the as-deposited zone, indicating that the interfacial bond is stronger than the interlayer bond inside the deposit. The fracture surfaces were analyzed using scanning electron microscopy (SEM) and energy disperse X-ray spectrometry (EDS). Results show that the tensile fracture failure of the as-deposited part is primarily affected by lack-of-fusion defects, carbide precipitation, and oxide particles inclusions, which causes premature failure of the deposit by deteriorating the mechanical properties and structural integrity.

Fabrication of Al/Mg/Al Composites via Accumulative Roll Bonding and Their Mechanical Properties

PubMed Central

Nie, Jinfeng; Liu, Mingxing; Wang, Fang; Zhao, Yonghao; Li, Yusheng; Cao, Yang; Zhu, Yuntian

2016-01-01

Al(1060)/Mg(AZ31)/Al(1060) multilayered composite was successfully produced using an accumulative roll bonding (ARB) process for up to four cycles at an elevated temperature (400 °C). The microstructure evolution of the composites and the bonding characteristics at the interfaces between Al and Mg layers with increasing ARB cycles were characterized through optical microscopy, field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM). It was found that the grains of Al and Mg layers were significantly refined and Al3Mg2 and Al12 Mg17 intermetallic compound layers formed at the Al/Mg bonding interfaces. The strength increased gradually and the ultimate tensile strength (UTS) reached a maximum value of about 240 MPa at the third pass. Furthermore, the strengthening mechanism of the composite was analyzed based on the fracture morphologies. PMID:28774072

Low-Cycle Fatigue Properties of P92 Ferritic-Martensitic Steel at Elevated Temperature

NASA Astrophysics Data System (ADS)

Zhang, Zhen; Hu, ZhengFei; Schmauder, Siegfried; Mlikota, Marijo; Fan, KangLe

2016-04-01

The low-cycle fatigue behavior of P92 ferritic-martensitic steel and the corresponding microstructure evolution at 873 K has been extensively studied. The test results of fatigue lifetime are consistent with the Coffin-Manson relationship over a range of controlled total strain amplitudes from 0.15 to 0.6%. The influence of strain amplitude on the fatigue crack initiation and growth has been observed using optical microscopy and scanning electron microscopy. The formation mechanism of secondary cracks is established according to the observation of fracture after fatigue process and there is an intrinsic relationship between striation spacing, current crack length, and strain amplitude. Transmission electron microscopy has been employed to investigate the microstructure evolution after fatigue process. It indicates the interaction between carbides and dislocations together with the formation of cell structure inhibits the cyclic softening. The low-angle sub-boundary elimination in the martensite is mainly caused by the cyclic stress.

Experimental Investigation of the Effect of Hydrogen on Fracture Toughness of 2.25Cr-1Mo-0.25V Steel and Welds after Annealing

PubMed Central

Song, Yan; Chai, Mengyu; Wu, Weijie; Liu, Yilun; Qin, Mu; Cheng, Guangxu

2018-01-01

Hydrogen embrittlement (HE) is a critical issue that hinders the reliability of hydrogenation reactors. Hence, it is of great significance to investigate the effect of hydrogen on fracture toughness of 2.25Cr-1Mo-0.25V steel and weld. In this work, the fracture behavior of 2.25Cr-1Mo-0.25V steel and welds was studied by three-point bending tests under hydrogen-free and hydrogen-charged conditions. The immersion charging method was employed to pre-charge hydrogen inside specimen and the fracture toughness of these joints was evaluated quantitatively. The microstructure and grain size of the specimens were observed by scanning electron microscopy (SEM) and by metallurgical microscopy to investigate the HE mechanisms. It was found that fracture toughness for both the base metal (BM) and the weld zone (WZ) significantly decreased under hydrogen-charged conditions due to the coexistence of the hydrogen-enhanced decohesion (HEDE) and hydrogen-enhanced localized plasticity (HELP) mechanisms. Moreover, the formation and growth of primary voids were observed in the BM, leading to a superior fracture toughness. In addition, the BM compared to the WZ shows superior resistance to HE because the finer grain size in the BM leads to a larger grain boundary area, thus distributing more of the diffusive hydrogen trapped in the grain boundary and reducing the hydrogen content. PMID:29584678

Mechanisms of hydrogen-assisted fracture in austenitic stainless steel welds.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Balch, Dorian K.; Sofronis, Petros; Somerday, Brian P.

2005-03-01

The objective of this study was to quantify the hydrogen-assisted fracture susceptibility of gas-tungsten arc (GTA) welds in the nitrogen-strengthened, austenitic stainless steels 21Cr-6Ni-9Mn (21-6-9) and 22Cr-13Ni-5Mn (22-13-5). In addition, mechanisms of hydrogen-assisted fracture in the welds were identified using electron microscopy and finite-element modeling. Elastic-plastic fracture mechanics experiments were conducted on hydrogen-charged GTA welds at 25 C. Results showed that hydrogen dramatically lowered the fracture toughness from 412 kJ/m{sup 2} to 57 kJ/m{sup 2} in 21-6-9 welds and from 91 kJ/m{sup 2} to 26 kJ/m{sup 2} in 22-13-5 welds. Microscopy results suggested that hydrogen served two roles in themore » fracture of welds: it promoted the nucleation of microcracks along the dendritic structure and accelerated the link-up of microcracks by facilitating localized deformation. A continuum finite-element model was formulated to test the notion that hydrogen could facilitate localized deformation in the ligament between microcracks. On the assumption that hydrogen decreased local flow stress in accordance with the hydrogen-enhanced dislocation mobility argument, the finite-element results showed that deformation was localized in a narrow band between two parallel, overlapping microcracks. In contrast, in the absence of hydrogen, the finite-element results showed that deformation between microcracks was more uniformly distributed.« less

Microstructural Evolution and Fracture Behavior of Friction-Stir-Welded Al-Cu Laminated Composites

NASA Astrophysics Data System (ADS)

Beygi, R.; Kazeminezhad, Mohsen; Kokabi, A. H.

2014-01-01

In this study, we attempt to characterize the microstructural evolution during friction stir butt welding of Al-Cu-laminated composites and its effect on the fracture behavior of the joint. Emphasis is on the material flow and particle distribution in the stir zone. For this purpose, optical microscopy and scanning electron microscopy (SEM) images, energy-dispersive spectroscopy EDS and XRD analyses, hardness measurements, and tensile tests are carried out on the joints. It is shown that intermetallic compounds exist in lamellas of banding structure formed in the advancing side of the welds. In samples welded from the Cu side, the banding structure in the advancing side and the hook formation in the retreating side determine the fracture behavior of the joint. In samples welded from the Al side, a defect is formed in the advancing side of the weld, which is attributed to insufficient material flow. It is concluded that the contact surface of the laminate (Al or Cu) with the shoulder of the FSW tool influences the material flow and microstructure of welds.

Solution and surface effects on plasma fibronectin structure

PubMed Central

1983-01-01

As assessed by electron microscopy, the reported shape of the plasma fibronectin molecule ranges from that of a compact particle to an elongated, rod-like structure. In this study, we evaluated the effects of solution and surface conditions on fibronectin shape. Freeze-dried, unstained human plasma fibronectin molecules deposited at pH 7.0-7.4 onto carbon films and examined by scanning transmission electron microscopy appeared relatively compact and pleiomorphic, with approximate average dimensions of 24 nm X 16 nm. Negatively stained molecules also had a similar shape but revealed greater detail in that we observed irregular, yarn-like structures. Glutaraldehyde-induced intramolecular cross-linking did not alter the appearance of plasma fibronectin. Molecules deposited at pH 2.8, pH 9.3, or after succinylation were less compact than those deposited at neutral pH. In contrast, fibronectin molecules sprayed onto mica surfaces at pH 7, rotary shadowed, and examined by transmission electron microscopy were elongated and nodular with a contour length of 120-130 nm. Sedimentation velocity experiments and electron microscopic observations indicate that fibronectin unfolds when it is succinylated, when the ionic strength is raised at pH 7, or when the pH is adjusted to 9.3 or 2.8. Greater unfolding is observed at pH 2.8 at low ionic strength (less than 0.01) compared with material at that pH in 0.15 M NaCl solution. We conclude that (a) the shape assumed by the fibronectin molecule can be strongly affected by solution conditions and by deposition onto certain surfaces; and that (b) the images of fibronectin seen by scanning transmission electron microscopy at neutral pH on carbon film are representative of molecules in physiologic solution. PMID:6417145

Fracture Toughness (KIC) of Lithography Based Manufactured Alumina Ceramic

NASA Astrophysics Data System (ADS)

Nindhia, T. G. T.; Schlacher, J.; Lube, T.

2018-04-01

Precision shaped ceramic components can be obtained by an emerging technique called Lithography based Ceramic Manufacturing (LCM). A green part is made from a slurry consisting of a ceramic powder in a photocurable binder with addition of dispersant and plasticizer. Components are built in a layer–by-layer way by exposing the desired cross- sections to light. The parts are subsequently sintered to their final density. It is a challenge to produce ceramic component with this method that yield the same mechanical properties in all direction. The fracture toughness (KIc) of of LCM-alumina (prepared at LITHOZ GmbH, Austria) was tested by using the Single-Edge-V-Notched Beam (SEVNB) method. Notches are made into prismatic bend-bars in all three direction X, Y and Z to recognize the value of fracture toughness of the material in all three directions. The microstructure was revealed with optical microscopy as well as Scanning Electron Microscopy (SEM). The results indicate that the fracture toughness in Y-direction has the highest value (3.10 MPam1/2) that is followed by the one in X-direction which is just a bit lower (2.90 MPam1/2). The Z-direction is found to have a similar fracture toughness (2.95 MPam1/2). This is supported by a homogeneous microstructure showing no hint of the layers used during production.

Fabrication of homogeneously dispersed graphene/Al composites by solution mixing and powder metallurgy

NASA Astrophysics Data System (ADS)

Zeng, Xiang; Teng, Jie; Yu, Jin-gang; Tan, Ao-shuang; Fu, Ding-fa; Zhang, Hui

2018-01-01

Graphene-reinforced aluminum (Al) matrix composites were successfully prepared via solution mixing and powder metallurgy in this study. The mechanical properties of the composites were studied using microhardness and tensile tests. Compared to the pure Al alloy, the graphene/Al composites showed increased strength and hardness. A tensile strength of 255 MPa was achieved for the graphene/Al composite with only 0.3wt% graphene, which has a 25% increase over the tensile strength of the pure Al matrix. Raman spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and transmission electron microscopy were used to investigate the morphologies, chemical compositions, and microstructures of the graphene and the graphene/Al composites. On the basis of fractographic evidence, a relevant fracture mechanism is proposed.

Sustained delivery of cytarabine-loaded vesicular phospholipid gels for treatment of xenografted glioma.

PubMed

Qi, Na; Cai, Cuifang; Zhang, Wei; Niu, Yantao; Yang, Jingyu; Wang, Lihui; Tian, Bin; Liu, Xiaona; Lin, Xia; Zhang, Yu; Zhang, Yan; He, Haibing; Chen, Kang; Tang, Xing

2014-09-10

This study described the development of vesicular phospholipid gels (VPGs) for sustained delivery of cytarabine (Ara-C) for the treatment of xenografted glioma. Ara-C-loaded VPGs in the state of a semisolid phospholipid dispersion looked like numerous vesicles tightly packing together under the freeze-fracture electron microscopy (FF-TEM), their release profiles displayed sustained drug release up to 384 h in vitro. The biodistribution of Ara-C in the rat brain showed that Ara-C-loaded VPGs could maintain therapeutic concentrations up to 5mm distance from the implantation site in brain tissue within 28 days. At the same time, fluorescence micrograph confirmed drug distribution in brain tissue visually. Furthermore, after single administration, Ara-C-loaded VPGs group significantly inhibited the U87-MG glioma growth in right flank in comparison with Ara-C solution (p<0.01). It was explained that the entrapped drug in VPGs could avoid degradation from cytidine deaminase and sustained release of drug from Ara-C-loaded VPGs could maintain the effective therapeutic levels for a long time around the tumor. In conclusion, Ara-C-loaded VPGs, with the properties of sustained release, high penetration capacity, nontoxicity and no shape restriction of the surgical cavity, are promising local delivery systems for post-surgical sustained chemotherapy against glioma. Copyright © 2014 Elsevier B.V. All rights reserved.

Ultrastructure of the extracellular matrix of bovine dura mater, optic nerve sheath and sclera.

PubMed Central

Raspanti, M; Marchini, M; Della Pasqua, V; Strocchi, R; Ruggeri, A

1992-01-01

The sclera, the outermost sheath of the optic nerve and the dura mater have been investigated histologically and ultrastructurally. Although these tissues appear very similar under the light microscope, being dense connective tissues mainly composed of collagen bundles and a limited amount of cells and elastic fibres, they exhibit subtle differences on electron microscopy. In the dura and sclera collagen appears in the form of large, nonuniform fibrils, similar to those commonly found in tendons, while in the optic nerve sheath the fibrils appear smaller and uniform, similar to those commonly observed in reticular tissues, vessel walls and skin. Freeze-fracture also reveals these fibrils to have different subfibrillar architectures, straight or helical, which correspond to 2 distinct forms of collagen fibril previously described (Raspanti et al. 1989). The other extracellular matrix components also vary with the particular collagen fibril structure. Despite their common embryological derivation, the dura mater, optic nerve sheath and sclera exhibit diversification of their extracellular matrix consistent with the mechanical loads to which these tissues are subjected. Our observations indicate that the outermost sheath of the optic nerve resembles the epineurium of peripheral nerves rather than the dura to which it is commonly likened. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 Fig. 6 Fig. 7 Fig. 8 Fig. 9 PMID:1295858

PsbS protein modulates non-photochemical chlorophyll fluorescence quenching in membranes depleted of photosystems.

PubMed

Ware, Maxwell A; Giovagnetti, Vasco; Belgio, Erica; Ruban, Alexander V

2015-11-01

Plants with varying levels of PsbS protein were grown on lincomycin. Enhanced levels of non-photochemical fluorescence quenching (NPQ) in over-expressers of the protein have been observed. This was accompanied by increased amplitude of the irreversible NPQ component, qI, previously considered to reflect mainly photoinhibition of PSII reaction centres (RCII). However, since RCIIs were largely absent the observed qI is likely to originate from the LHCII antenna. In chloroplasts of over-expressers of PsbS grown on lincomycin an abnormally large NPQ (∼7) was characterised by a 0.34 ns average chlorophyll fluorescence lifetime. Yet the lifetime in the Fm state was similar to that of wild-type plants. 77K fluorescence emission spectra revealed a specific 700 nm peak typical of LHCII aggregates as well as quenching of the PSI fluorescence at 730 nm. The aggregated state manifested itself as a clear change in the distance between LHCII complexes detected by freeze-fracture electron microscopy. Grana thylakoids in the quenched state revealed 3 times more aggregated LHCII particles compared to the dark-adapted state. Overall, the results directly demonstrate the importance of LHCII aggregation in the NPQ mechanism and show that the PSII supercomplex structure plays no role in formation of the observed quenching. Copyright © 2015 Elsevier B.V. All rights reserved.

The effect of Al intermediate layer on thermal resistance of EB-PVD yttria-stabilized zirconia coatings on titanium substrate

NASA Astrophysics Data System (ADS)

Panin, Alexey; Panin, Victor; Kazachenok, Marina; Shugurov, Artur; Sinyakova, Elena; Martynov, Sergey; Rusyaev, Andrey; Kasterov, Artur

2017-12-01

The yttria-stabilized zirconia coatings sprayed on titanium substrates by the electron beam physical vapor deposition were subjected to thermal annealing in air at 1000°C for 1, 30 and 60 min. The delamination and fracture of the coatings are studied by the scanning electron microscopy and X-ray diffraction. It is shown that a magnetron sputtered Al interlayer between the coating and the substrate considerably improves the thermal resistance of ceramic coatings.

Isolation of a Soluble Component of Plasmodium berghei Which Induces Immunity in Rats

PubMed Central

Grothaus, G. D.; Kreier, J. P.

1980-01-01

Soluble material was obtained from sonically freed plasmodiae by three procedures. Two procedures, cryo-impacting and freeze-thawing, were evaluated for their ability to disrupt the parasites and release soluble material. The soluble materials obtained by these procedures were compared to materials washed from the surfaces of sonically freed parasites. Between 35 and 40% of the total parasite protein was solubilized by freeze-thawing or cryo-impacting. One cycle of freeze-thawing released nearly as much protein as could be released by this method, and additional cycles of freeze-thawing had little additional effect. Cryo-impacting solubilized only a small amount of protein in addition to that which was released by the cycle of freeze-thawing inherent in the procedure. Reductions in the packed cell volume of the material remaining after freeze-thawing or cryo-impacting indicate that the insoluble fragments are broken into smaller pieces as treatment is extended. Electron microscopy of 30-s cryo-impacted and three-times freeze-thawed parasites revealed membrane fragments similar in appearance. Patterns obtained by polyacrylamide gel electrophoresis of the soluble material from freeze-thawed and cryo-impacted parasites were also similar, and approximately 13 protein bands were demonstrated. The material washed from the surfaces of the free parasites, on the other hand, resolved into only two to four major bands on the gel columns. In immunization studies, the soluble and insoluble fractions obtained by freeze-thawing or cryo-impacting and the material washed from the surfaces of the parasites all stimulated a protective immune response. On the basis of the amount of protein required to stimulate roughly comparable immunity, the soluble fraction obtained by freeze-thawing or cryo-impacting free parasites was about twice as potent an immunogen as was the insoluble fraction. The material obtained by gentle washing of the freed parasites was approximately 20 times as potent an immunogen as were the freed parasites and about 7 times as potent as the soluble material obtained by freeze-thawing or cryo-impacting. Images Fig. 1 Fig. 2 PMID:6991439

Electron microscopy of the amphibian model systems Xenopus laevis and Ambystoma mexicanum.

PubMed

Kurth, Thomas; Berger, Jürgen; Wilsch-Bräuninger, Michaela; Kretschmar, Susanne; Cerny, Robert; Schwarz, Heinz; Löfberg, Jan; Piendl, Thomas; Epperlein, Hans H

2010-01-01

In this chapter we provide a set of different protocols for the ultrastructural analysis of amphibian (Xenopus, axolotl) tissues, mostly of embryonic origin. For Xenopus these methods include: (1) embedding gastrulae and tailbud embryos into Spurr's resin for TEM, (2) post-embedding labeling of methacrylate (K4M) and cryosections through adult and embryonic epithelia for correlative LM and TEM, and (3) pre-embedding labeling of embryonic tissues with silver-enhanced nanogold. For the axolotl (Ambystoma mexicanum) we present the following methods: (1) SEM of migrating neural crest (NC) cells; (2) SEM and TEM of extracellular matrix (ECM) material; (3) Cryo-SEM of extracellular matrix (ECM) material after cryoimmobilization; and (4) TEM analysis of hyaluronan using high-pressure freezing and HABP labeling. These methods provide exemplary approaches for a variety of questions in the field of amphibian development and regeneration, and focus on cell biological issues that can only be answered with fine structural imaging methods, such as electron microscopy. Copyright © 2010 Elsevier Inc. All rights reserved.

Green synthesis of silver nanoparticles from Moringa oleifera leaf extracts and its antimicrobial potential

NASA Astrophysics Data System (ADS)

Moodley, Jerushka S.; Babu Naidu Krishna, Suresh; Pillay, Karen; Sershen; Govender, Patrick

2018-03-01

In this study we report on the synthesis of silver nanoparticles (AgNPs) from the leaf extracts of Moringa oleifera using sunlight irradiation as primary source of energy, and its antimicrobial potential. Silver nanoparticle formation was confirmed by surface plasmon resonance at 450 nm and 440 nm, respectively for both fresh and freeze-dried leaf samples. Crystanality of AgNPs was confirmed by transmission electron microscopy, scanning electron microscopy with energy dispersive x-ray spectroscopy and Fourier transform infrared (FTIR) spectroscopy analysis. FTIR spectroscopic analysis suggested that flavones, terpenoids and polysaccharides predominate and are primarily responsible for the reduction and subsequent capping of AgNPs. X-ray diffraction analysis also demonstrated that the size range of AgNPs from both samples exhibited average diameters of 9 and 11 nm, respectively. Silver nanoparticles showed antimicrobial activity on both bacterial and fungal strains. The biosynthesised nanoparticle preparations from M. oleifera leaf extracts exhibit potential for application as broad-spectrum antimicrobial agents.

Hierarchical microstructures in CZT

NASA Astrophysics Data System (ADS)

Sundaram, S. K.; Henager, C. H.; Edwards, D. J.; Schemer-Kohrn, A. L.; Bliss, M.; Riley, B. R.; Toloczko, M. B.; Lynn, K. G.

2011-10-01

Advanced characterization tools, such as electron backscatter diffraction and transmitted IR microscopy, are being applied to study critical microstructural features and orientation relations in as-grown CZT crystals to aid in understanding the relation between structure and properties in radiation detectors. Even carefully prepared single crystals of CZT contain regions of slight misorientation, Te-particles, and dislocation networks that must be understood for more accurate models of detector response. This paper describes initial research at PNNL into the hierarchy of microstructures observed in CZT grown via the vertical gradient freeze or vertical Bridgman method at PNNL and WSU.

Experimental Study and Fractal Analysis on the Anisotropic Performance of Explosively Welded Interfaces of 304 Stainless Steel/245 Carbon Steel

NASA Astrophysics Data System (ADS)

Fu, Yanshu; Qiu, Yaohui; Li, Yulong

2018-03-01

The mechanical anisotropy of an explosive welding composite plate made of 304 stainless steel/245 steel was studied through shear experiments performed on explosively welded wavy interfaces along several orientation angles. The results indicated that the strength and the fracture energy of samples significantly varied with the orientation angles. The fracture surfaces of all samples were observed using a scanning electron microscope and through three-dimensional structure microscopy. The periodic features of all the fracture surfaces were clearly shown in different fracture modes. The fractal dimension of the fracture surfaces was calculated based on the fractal geometry by the box-counting method in MATLAB. The cohesive element model was used to analyze the fracture energy according to the physical dependence of the fractal dimension on thermodynamic entropy and interface separation energy. The fracture energy was an exponential function of the fractal dimension value, which was in good agreement with the experimental results. All results were validated for effective use in the application of anisotropy analysis to the welded interface and structural optimization of explosively welded composite plates.

Comparison of Intralaminar and Interlaminar Mode-I Fracture Toughness of Unidirectional IM7/8552 Graphite/Epoxy Composite

NASA Technical Reports Server (NTRS)

Czabaj, Michael W.; Ratcliffe, James

2012-01-01

The intralaminar and interlaminar mode-I fracture-toughness of a unidirectional IM7/8552 graphite/epoxy composite were measured using compact tension (CT) and double cantilever beam (DCB) test specimens, respectively. Two starter crack geometries were considered for both the CT and DCB specimen configurations. In the first case, starter cracks were produced by 12.5 micron thick, Teflon film inserts. In the second case, considerably sharper starter cracks were produced by fatigue precracking. For each specimen configuration, use of the Teflon film starter cracks resulted in initially unstable crack growth and artificially high initiation fracture-toughness values. Conversely, specimens with fatigue precracks exhibited stable growth onset and lower initiation fracture toughness. For CT and DCB specimens with fatigue precracks, the intralaminar and interlaminar initiation fracture toughnesses were approximately equal. However, during propagation, the CT specimens exhibited more extensive fiber bridging, and rapidly increasing R-curve behavior as compared to the DCB specimens. Observations of initiation and propagation of intralaminar and interlaminar fracture, and the measurements of fracture toughness, were supported by fractographic analysis using scanning electron microscopy.

Synchrotron quantification of fracturing during maturation of shales

NASA Astrophysics Data System (ADS)

Figueroa Pilz, Fernando; Fauchille, Anne-Laure; Dowey, Patrick; Courtois, Loic; Bay, Brian; Ma, Lin; Taylor, Kevin; Mecklenburgh, Julian; Lee, Peter

2017-04-01

To understand both the hydrocarbon migration within and from shale rocks, and during hydraulic fracturing, is needed to evaluate and predict its environmental footprint. As a consequence, the time characterization of fracture networks in shale is particularly important. Time resolved synchrotron X-ray tomography was used to quantify the initiation and propagation of fractures during the simulated maturation of an organic-rich Kimmeridge Clay shale from the µm to mm scales. Scanning electron microscopy (SEM) observations were performed before and after maturation in order to compare the microstructure evolution and better understand the fracture location. Fracture and strain development during heating was quantified in 3D by Digital Volume Correlation (DVC) (Bay et al., 1999). The combination of DVC, X-Ray tomography and SEM obtained direct 4D strain measurements of the anisotropic mechanical behaviour of Kimmeridge shale with the temperature during an accelerated thermal maturation (Figueroa Pilz et al.). Such a combination has rarely been investigated in 4D at these scales in the past. In the study conditions, the results demonstrated the anisotropy in thermal expansion and the aperture fracture pathways through organic matter and clay matrix.

Experimental Study and Fractal Analysis on the Anisotropic Performance of Explosively Welded Interfaces of 304 Stainless Steel/245 Carbon Steel

NASA Astrophysics Data System (ADS)

Fu, Yanshu; Qiu, Yaohui; Li, Yulong

2018-05-01

The mechanical anisotropy of an explosive welding composite plate made of 304 stainless steel/245 steel was studied through shear experiments performed on explosively welded wavy interfaces along several orientation angles. The results indicated that the strength and the fracture energy of samples significantly varied with the orientation angles. The fracture surfaces of all samples were observed using a scanning electron microscope and through three-dimensional structure microscopy. The periodic features of all the fracture surfaces were clearly shown in different fracture modes. The fractal dimension of the fracture surfaces was calculated based on the fractal geometry by the box-counting method in MATLAB. The cohesive element model was used to analyze the fracture energy according to the physical dependence of the fractal dimension on thermodynamic entropy and interface separation energy. The fracture energy was an exponential function of the fractal dimension value, which was in good agreement with the experimental results. All results were validated for effective use in the application of anisotropy analysis to the welded interface and structural optimization of explosively welded composite plates.

Tritium and decay helium effects on the fracture toughness properties of types 316L, 304L and 21Cr-6Ni-9Mn stainless steels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Morgan, M.J.; Tosten, M.H

1994-10-01

J-integral fracture mechanics techniques and electron microscopy observations were used to investigate the effects of tritium and its radioactive decay product, {sup 3}He, on Types 316L, 304L and 21Cr-6Ni-9Mn stainless steels. Tritium-exposed-and-aged steels had lower fracture-toughness values and shallower sloped crack-growth-resistance curves than unexposed steels. Both fracture-toughness parameters decreased with increasing concentrations of {sup 3}He. The fracture-toughness reductions were accompanied by a change in fracture mode from microvoid-nucleation-and-growth processes in control samples to grain-and-twin-boundary fracture in tritium-charged-and-aged samples. Type 316L stainless steel had the highest fracture-toughness values and Type 21Cr-6Ni-9Mn had the lowest. Samples containing {sup 3}He but degassed ofmore » tritium had fracture toughness properties that were similar to uncharged samples. The results indicate that helium bubbles enhance the embrittlement effects of hydrogen by affecting the deformation properties and by increasing localized hydrogen concentrations through trapping effects.« less

[Reparative Osteogenesis and Angiogenesis in Low Intensity Electromagnetic Radiation of Ultra-High Frequency].

PubMed

Iryanov, Y M; Kiryanov, N A

2015-01-01

Non-drug correction of reparative bone tissue regeneration in different pathological states - one of the most actual problems of modern medicine. Our aim was to conduct morphological analysis of the influence of electromagnetic radiation of ultra-high frequency and low intensity on reparative osteogenesis and angiogenesis in fracture treatment under transosseous osteosynthesis. A controlled nonrandomized study was carried out. In the experiment conducted on rats we modeled tibial fracture with reposition and fixation of the bone fragments both in control and experimental groups. In the animals of the experimental group the fracture zone was exposed to low intensity electromagnetic radiation of ultra-high frequency. Exposure simulation was performed in the control group. The operated bones were examined using radiography, light and electronic microscopy, X-ray electronic probe microanalysis. It has been established that electromagnetic radiation of ultra-high frequency sessions in fracture treatment stimulate secretory activity and degranulation of mast cells, produce microcirculatory bed vascular permeability increase, endotheliocyte migration phenotype expression, provide endovascular endothelial outgrowth formation, activate reparative osteogenesis and angiogenesis while fracture reparation becomes the one of the primary type. The full periosteal, intermediary and intraosteal bone union was defined in 28 days. Among the therapeutic benefits of electromagnetic radiation of ultra-high frequency in fracture treatment we can detect mast cell secretorv activity stimulation and endovascular anziozenesis activation.

Ultrastructure of the mycangium of the southern pine beetle, Dendroctonus frontalis (Coleoptera: Curculionidae, Scolytinae): complex morphology for complex interactions

Treesearch

Cetin Yuceer; Chuan-Yu Hsu; Nadir Erbilgin; Kier D. Klepzig

2011-01-01

The southern pine beetle (SPB) (Dendroctonus frontalis Zimmermann) is the most economically important pest of southern pine forests. Beetles carry fungal cells within specialised cuticular structures, called mycangia. Little is known about the mycangia ultrastructure or function. We used cryo-fracturing and scanning electron microscopy to examine the ultrastructural...

High damage tolerance of electrochemically lithiated silicon

PubMed Central

Wang, Xueju; Fan, Feifei; Wang, Jiangwei; Wang, Haoran; Tao, Siyu; Yang, Avery; Liu, Yang; Beng Chew, Huck; Mao, Scott X.; Zhu, Ting; Xia, Shuman

2015-01-01

Mechanical degradation and resultant capacity fade in high-capacity electrode materials critically hinder their use in high-performance rechargeable batteries. Despite tremendous efforts devoted to the study of the electro–chemo–mechanical behaviours of high-capacity electrode materials, their fracture properties and mechanisms remain largely unknown. Here we report a nanomechanical study on the damage tolerance of electrochemically lithiated silicon. Our in situ transmission electron microscopy experiments reveal a striking contrast of brittle fracture in pristine silicon versus ductile tensile deformation in fully lithiated silicon. Quantitative fracture toughness measurements by nanoindentation show a rapid brittle-to-ductile transition of fracture as the lithium-to-silicon molar ratio is increased to above 1.5. Molecular dynamics simulations elucidate the mechanistic underpinnings of the brittle-to-ductile transition governed by atomic bonding and lithiation-induced toughening. Our results reveal the high damage tolerance in amorphous lithium-rich silicon alloys and have important implications for the development of durable rechargeable batteries. PMID:26400671

High damage tolerance of electrochemically lithiated silicon

NASA Astrophysics Data System (ADS)

Wang, Xueju; Fan, Feifei; Wang, Jiangwei; Wang, Haoran; Tao, Siyu; Yang, Avery; Liu, Yang; Beng Chew, Huck; Mao, Scott X.; Zhu, Ting; Xia, Shuman

2015-09-01

Mechanical degradation and resultant capacity fade in high-capacity electrode materials critically hinder their use in high-performance rechargeable batteries. Despite tremendous efforts devoted to the study of the electro-chemo-mechanical behaviours of high-capacity electrode materials, their fracture properties and mechanisms remain largely unknown. Here we report a nanomechanical study on the damage tolerance of electrochemically lithiated silicon. Our in situ transmission electron microscopy experiments reveal a striking contrast of brittle fracture in pristine silicon versus ductile tensile deformation in fully lithiated silicon. Quantitative fracture toughness measurements by nanoindentation show a rapid brittle-to-ductile transition of fracture as the lithium-to-silicon molar ratio is increased to above 1.5. Molecular dynamics simulations elucidate the mechanistic underpinnings of the brittle-to-ductile transition governed by atomic bonding and lithiation-induced toughening. Our results reveal the high damage tolerance in amorphous lithium-rich silicon alloys and have important implications for the development of durable rechargeable batteries.

Microstructure and Precipitate's Characterization of the Cu-Ni-Si-P Alloy

NASA Astrophysics Data System (ADS)

Zhang, Yi; Tian, Baohong; Volinsky, Alex A.; Sun, Huili; Chai, Zhe; Liu, Ping; Chen, Xiaohong; Liu, Yong

2016-04-01

Microstructure of the Cu-Ni-Si-P alloy was investigated by transmission electron microscopy (TEM). The alloy had 551 MPa tensile strength, 226 HV hardness, and 36% IACS electrical conductivity after 80% cold rolling and aging at 450 °C for 2 h. Under the same aging conditions, but without the cold rolling, the strength, hardness, and electrical conductivity were 379 MPa, 216 HV, and 32% IACS, respectively. The precipitates identified by TEM characterization were δ-Ni2Si. Some semi-coherent spherical precipitates with a typical coffee bean contrast were found after aging for 48 h at 450 °C. The average diameter of the observed semi-coherent precipitates is about 5 nm. The morphology of the fracture surface was observed by scanning electron microscopy. All samples showed typical ductile fracture. The addition of P refined the grain size and increased the nucleation rate of the precipitates. The precipitated phase coarsening was inhibited by the small additions of P. After aging, the Cu-Ni-Si-P alloy can gain excellent mechanical properties with 804 MPa strength and 49% IACS conductivity. This study aimed to optimize processing conditions of the Cu-Ni-Si-P alloys.

Annealing to optimize the primary drying rate, reduce freezing-induced drying rate heterogeneity, and determine T(g)' in pharmaceutical lyophilization.

PubMed

Searles, J A; Carpenter, J F; Randolph, T W

2001-07-01

In a companion paper we show that the freezing of samples in vials by shelf-ramp freezing results in significant primary drying rate heterogeneity because of a dependence of the ice crystal size on the nucleation temperature during freezing.1 The purpose of this study was to test the hypothesis that post-freezing annealing, in which the product is held at a predetermined temperature for a specified duration, can reduce freezing-induced heterogeneity in sublimation rates. In addition, we test the impact of annealing on primary drying rates. Finally, we use the kinetics of relaxations during annealing to provide a simple measurement of T(g)', the glass transition temperature of the maximally freeze-concentrated amorphous phase, under conditions and time scales most appropriate for industrial lyophilization cycles. Aqueous solutions of hydroxyethyl starch (HES), sucrose, and HES:sucrose were either frozen by placement on a shelf while the temperature was reduced ("shelf-ramp frozen") or by immersion into liquid nitrogen. Samples were then annealed for various durations over a range of temperatures and partially lyophilized to determine the primary drying rate. The morphology of fully dried liquid nitrogen-frozen samples was examined using scanning electron microscopy. Annealing reduced primary drying rate heterogeneity for shelf-ramp frozen samples, and resulted in up to 3.5-fold increases in the primary drying rate. These effects were due to increased ice crystal sizes, simplified amorphous structures, and larger and more numerous holes on the cake surface of annealed samples. Annealed HES samples dissolved slightly faster than their unannealed counterparts. Annealing below T(g)' did not result in increased drying rates. We present a simple new annealing-lyophilization method of T(g)' determination that exploits this phenomenon. It can be carried out with a balance and a freeze-dryer, and has the additional advantage that a large number of candidate formulations can be evaluated simultaneously.

Rapid Tempering of Martensitic Stainless Steel AISI420: Microstructure, Mechanical and Corrosion Properties

NASA Astrophysics Data System (ADS)

Abbasi-Khazaei, Bijan; Mollaahmadi, Akbar

2017-04-01

In this research, the effect of rapid tempering on the microstructure, mechanical properties and corrosion resistance of AISI 420 martensitic stainless steel has been investigated. At first, all test specimens were austenitized at 1050 °C for 1 h and tempered at 200 °C for 1 h. Then, the samples were rapidly reheated by a salt bath furnace in a temperature range from 300 to 1050 °C for 2 min and cooled in air. The tensile tests, impact, hardness and electrochemical corrosion were carried out on the reheated samples. Scanning electron microscopy was used to study the microstructure and fracture surface. To investigate carbides, transmission electron microscopy and also scanning electron microscopy were used. X-ray diffraction was used for determination of the retained austenite. The results showed that the minimum properties such as the tensile strength, impact energy, hardness and corrosion resistance were obtained at reheating temperature of 700 °C. Semi-continuous carbides in the grain boundaries were seen in this temperature. Secondary hardening phenomenon was occurred at reheating temperature of 500 °C.

Membrane interactions between secretion granules and plasmalemma in three exocrine glands

PubMed Central

Tanaka, Y; De Camilli, P; Meldolesi, J

1980-01-01

Three types of membrane interactions were studied in three exocrine systems (the acinar cells of the rat parotid, rat lacrimal gland, and guinea pig pancrease) by freeze- fracture and thin-section electron microscopy: exocytosis, induced in vivo by specific pharmacological stimulations; the mutual apposition of secretory granule membranes in the intact cell; membrane appositions induced in vitro by centrifugation of the isolated granules. In all three glandular cells, the distribution of intramembrane particles (IMP) on the fracture faces of the luminal plasmagranule membrane particles (IMP) on the fracture faces of the lumenal plasmalemma appeared random before stimulation. However, after injection of secretagogues, IMP were rapidly clearly from the areas of granule- plasmalemma apposition in the parotid cells and, especially, in lacrimocytes. In the latter, the cleared areas appeared as large bulges toward the lumen, whereas in the parotid they were less pronounced. Exocytotic openings were usually large and the fracture faces of their rims were covered with IMP. In contrast, in stimulated pancreatic acinar cells, the IMP distribution remained apparently random after stimulation. Exocytoses were established through the formation of narrown necks, and no images which might correspond to early stages of membrane fusion were revealed. Within the cytoplasm of parotid and lacrimal cells (but not in the pancreas), both at rest and after stimulation, secretion granules were often closely apposed by means of flat, circular areas, also devoid of IMP. In thin sections, the images corresponding to IMP-free areas were close granule-granule and granule-plasmalemma appositions, sometimes with focal merging of the membrane outer layers to yield pentalaminar structures. Isolated secretion granules were forced together in vitro by centrifugation. Under these conditions, increasing the centrifugal force from 1,600 to 50,000 g for 10 min resulted in a progressive, statistically significant increase of the frequency of IMP-free flat appositions between parotid granules. In contrast, no such areas were seen between freeze-fractured pancreatic granules, although some focal pentalaminar appositions appeared in section after centrifugation at 50 and 100,000 g for 10 min. On the basis of the observation that, in secretory cells, IMP clearing always develops in deformed membrane areas (bulges, depressions, flat areas), it is suggested that it might result from the forced mechanical apposition of the interacting membranes. This might be a preliminary process not sufficient to initiate fusion. In the pancreas, IMP clearing could occur over surface areas too small to be detected. In stimulated parotid and lacrimal glands they were exceptional. These structures were either attached at the sites of continuity between granule and plasma membranes, or free in the acinar lumen, with a preferential location within exocytotic pockets or in their proximity. Experiments designed to investigate the nature of these blisters and vesicles revealed that they probably arise artifactually during glutaraldehyde fixation. In fact, (a) they were large and numerous in poorly fixed samples but were never observed in thin sections of specimens fixed in one step with glutaraldehyde and OsO(4); and (b) no increase in concentration of phospholipids was observed in the parotid saliva and pancreatic juice after stimulation of protein discharge, as was to be expected if release of membrane material were occurring after exocytosis. PMID:7380885

Aging and loading rate effects on the mechanical behavior of equine bone

NASA Astrophysics Data System (ADS)

Kulin, Robb M.; Jiang, Fengchun; Vecchio, Kenneth S.

2008-06-01

Whether due to a sporting accident, high-speed impact, fall, or other catastrophic event, the majority of clinical bone fractures occur under dynamic loading conditions. However, although extensive research has been performed on the quasi-static fracture and mechanical behavior of bone to date, few high-quality studies on the fracture behavior of bone at high strain rates have been performed. Therefore, many questions remain regarding the material behavior, including not only the loading-rate-dependent response of bone, but also how this response varies with age. In this study, tests were performed on equine femoral bone taken post-mortem from donors 6 months to 28 years of age. Quasi-static and dynamic tests were performed to determine the fracture toughness and compressive mechanical behavior as a function of age at varying loading rates. Fracture paths were then analyzed using scanning confocal and scanning-electron microscopy techniques to assess the role of various microstructural features on toughening mechanisms.

Effect of strain rate and notch geometry on tensile properties and fracture mechanism of creep strength enhanced ferritic P91 steel

NASA Astrophysics Data System (ADS)

Pandey, Chandan; Mahapatra, M. M.; Kumar, Pradeep; Saini, N.

2018-01-01

Creep strength enhanced ferritic (CSEF) P91 steel were subjected to room temperature tensile test for quasi-static (less than 10-1/s) strain rate by using the Instron Vertical Tensile Testing Machine. Effect of different type of notch geometry, notch depth and angle on mechanical properties were also considered for different strain rate. In quasi-static rates, the P91 steel showed a positive strain rate sensitivity. On the basis of tensile data, fracture toughness of P91 steel was also calculated numerically. For 1 mm notch depth (constant strain rate), notch strength and fracture toughness were found to be increased with increase in notch angle from 45° to 60° while the maximum value attained in U-type notch. Notch angle and notch depth has found a minute effect on P91 steel strength and fracture toughness. The fracture surface morphology was studied by field emission scanning electron microscopy (FESEM).

Immersion Freezing of Coal Combustion Ash Particles from the Texas Panhandle

NASA Astrophysics Data System (ADS)

Whiteside, C. L.; Tobo, Y.; Mulamba, O.; Brooks, S. D.; Mirrielees, J.; Hiranuma, N.

2017-12-01

Coal combustion aerosol particles contribute to the concentrations of ice-nucleating particles (INPs) in the atmosphere. Especially, immersion freezing can be considered as one of the most important mechanisms for INP formation in supercooled tropospheric clouds that exist at temperatures between 0°C and -38°C. The U.S. contains more than 550 operating coal-burning plants consuming 7.2 x 108 metric tons of coal (in 2016) to generate a total annual electricity of >2 billion MW-h, resulting in the emission of at least 4.9 x 105 metric tons of PM10 (particulate matter smaller than 10 µm in diameter). In Texas alone, 19 combustion plants generate 0.15 billion MW-h electricity and >2.4 x 104 metric tons of PM10. Here we present the immersion freezing behavior of combustion fly ash and bottom ash particles collected in the Texas Panhandle region. Two types of particulate samples, namely <45 µm sieved bottom ash (B_Ash_TX_PH) and <45 µm sieved fly ash (F_Ash_TX_PH), were prepared. Afterwards, their immersion freezing abilities were measured using the Cryogenic Refrigerator Applied to Freezing Test (CRAFT) system covering the heterogeneous freezing temperature down to -30 °C. The results were generated and are reported through two metrics, frozen fraction, ffrozen(T), and ice nucleation active site density per unit mass, nm(T) as a function of temperature. Our preliminary results show that an onset increase in ffrozen(T) for B_Ash_TX_PH (ffrozen) occurred as high as at -15°C, whereas the onset for F_Ash_TX_PH is at -18°C. Secondly, B_Ash_TX_PH exhibited a higher nm(-20 °C) of 105 g-1 than that of F_Ash_TX_PH ( 5 x 103 g-1). On the other hand, previous studies on different combustion ash samples have reported that the opposite trend (i.e., ice nucleation efficiency of fly ash is greater than that of bottom ash; Grawe et al., 2016, ACP; Umo et al., 2015, ACP). We will discuss possible reasons for the observed differences. In addition, the results of complementary physico-chemical analyses via X-ray diffraction technique, Raman microscopy and scanning electron microscopy on both ash types will also be presented to relate the crystallographic and chemical properties to their ice nucleation abilities.

Lamellae spatial distribution modulates fracture behavior and toughness of african pangolin scales

DOE PAGES

Chon, Michael J.; Daly, Matthew; Wang, Bin; ...

2017-06-10

Pangolin scales form a durable armor whose hierarchical structure offers an avenue towards high performance bio-inspired materials design. In this paper, the fracture resistance of African pangolin scales is examined using single edge crack three-point bend fracture testing in order to understand toughening mechanisms arising from the structures of natural mammalian armors. In these mechanical tests, the influence of material orientation and hydration level are examined. The fracture experiments reveal an exceptional fracture resistance due to crack deflection induced by the internal spatial orientation of lamellae. An order of magnitude increase in the measured fracture resistance due to scale hydration,more » reaching up to ~ 25 kJ/m 2 was measured. Post-mortem analysis of the fracture samples was performed using a combination of optical and electron microscopy, and X-ray computerized tomography. Interestingly, the crack profile morphologies are observed to follow paths outlined by the keratinous lamellae structure of the pangolin scale. Most notably, the inherent structure of pangolin scales offers a pathway for crack deflection and fracture toughening. Finally, the results of this study are expected to be useful as design principles for high performance biomimetic applications.« less

Lamellae spatial distribution modulates fracture behavior and toughness of african pangolin scales.

PubMed

Chon, Michael J; Daly, Matthew; Wang, Bin; Xiao, Xianghui; Zaheri, Alireza; Meyers, Marc A; Espinosa, Horacio D

2017-12-01

Pangolin scales form a durable armor whose hierarchical structure offers an avenue towards high performance bio-inspired materials design. In this study, the fracture resistance of African pangolin scales is examined using single edge crack three-point bend fracture testing in order to understand toughening mechanisms arising from the structures of natural mammalian armors. In these mechanical tests, the influence of material orientation and hydration level are examined. The fracture experiments reveal an exceptional fracture resistance due to crack deflection induced by the internal spatial orientation of lamellae. An order of magnitude increase in the measured fracture resistance due to scale hydration, reaching up to ~ 25kJ/m 2 was measured. Post-mortem analysis of the fracture samples was performed using a combination of optical and electron microscopy, and X-ray computerized tomography. Interestingly, the crack profile morphologies are observed to follow paths outlined by the keratinous lamellae structure of the pangolin scale. Most notably, the inherent structure of pangolin scales offers a pathway for crack deflection and fracture toughening. The results of this study are expected to be useful as design principles for high performance biomimetic applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effects of electropolishing surface treatment on the cyclic fatigue resistance of BioRace nickel-titanium rotary instruments.

PubMed

Lopes, Hélio P; Elias, Carlos N; Vieira, Victor T L; Moreira, Edson J L; Marques, Raquel V L; de Oliveira, Julio C Machado; Debelian, Gilberto; Siqueira, José F

2010-10-01

This study evaluated the influence of electropolishing surface treatment on the number of cycles to fracture of BioRace rotary nickel-titanium endodontic instruments. BioRace size BR5C instruments with or without electropolishing surface treatment were used in an artificial curved canal under rotational speed of 300 rpm until fracture. Fractured surfaces and the helical shafts of fractured instruments were analyzed by scanning electron microscopy (SEM). Polished instruments displayed a significantly higher number of cycles to fracture when compared with nonpolished instruments (P < .001). Actually, the number of cycles to fracture of a polished BR5C instrument was 124% higher than that of a nonpolished instrument. SEM analysis showed that the fractured surface of both polished and nonpolished BR5C instruments had ductile morphologic characteristics. Evaluation of the separated fragments after cyclic fatigue testing showed the presence of microcracks near the fracture surface. Polished instruments exhibited fine cracks that assumed an irregular path (zigzag crack pattern), whereas nonpolished instruments showed cracks running along the machining grooves. Electropolishing surface treatment of BioRace endodontic instruments significantly increased the cyclic fatigue resistance. Copyright © 2010 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Cyclic fatigue resistance of ProTaper Universal instruments when subjected to static and dynamic tests.

PubMed

Lopes, Hélio P; Britto, Izabelle M O; Elias, Carlos N; Machado de Oliveira, Julio C; Neves, Mônica A S; Moreira, Edson J L; Siqueira, José F

2010-09-01

This study evaluated the number of cycles to fracture of ProTaper Universal S2 instruments when subjected to static and dynamic cyclic fatigue tests. ProTaper Universal S2 instruments were used until fracture in an artificial curved canal under rotational speed of 300 rpm in either a static or a dynamic test model. Afterward, the length of the fractured segments was measured and fractured surfaces and helical shafts analyzed by scanning electron microscopy (SEM). The number of cycles to fracture was significantly increased when instruments were tested in the dynamic model (P<.001). Instrument separation occurred at the point of maximum flexure within the artificial canals, i.e., the midpoint of the curved canal segment. SEM analysis revealed that fractured surfaces exhibited characteristics of the ductile mode. Plastic deformation was not observed in the helical shaft of fractured instruments. The number of cycles to fracture ProTaper Universal S2 instruments significantly increased with the use of instruments in a dynamic cyclic fatigue test compared with a static model. These findings reinforce the need for performing continuous pecking motions during rotary instrumentation of curved root canals. Copyright (c) 2010 Mosby, Inc. All rights reserved.

Lamellae spatial distribution modulates fracture behavior and toughness of african pangolin scales

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chon, Michael J.; Daly, Matthew; Wang, Bin

Pangolin scales form a durable armor whose hierarchical structure offers an avenue towards high performance bio-inspired materials design. In this paper, the fracture resistance of African pangolin scales is examined using single edge crack three-point bend fracture testing in order to understand toughening mechanisms arising from the structures of natural mammalian armors. In these mechanical tests, the influence of material orientation and hydration level are examined. The fracture experiments reveal an exceptional fracture resistance due to crack deflection induced by the internal spatial orientation of lamellae. An order of magnitude increase in the measured fracture resistance due to scale hydration,more » reaching up to ~ 25 kJ/m 2 was measured. Post-mortem analysis of the fracture samples was performed using a combination of optical and electron microscopy, and X-ray computerized tomography. Interestingly, the crack profile morphologies are observed to follow paths outlined by the keratinous lamellae structure of the pangolin scale. Most notably, the inherent structure of pangolin scales offers a pathway for crack deflection and fracture toughening. Finally, the results of this study are expected to be useful as design principles for high performance biomimetic applications.« less

Electronically transparent graphene replicas of diatoms: a new technique for the investigation of frustule morphology

PubMed Central

Pan, Zhengwei; Lerch, Sarah J. L.; Xu, Liang; Li, Xufan; Chuang, Yen-Jun; Howe, Jane Y.; Mahurin, Shannon M.; Dai, Sheng; Hildebrand, Mark

2014-01-01

The morphogenesis of the silica cell walls (called frustules) of unicellular algae known as diatoms is one of the most intriguing mysteries of the diatoms. To study frustule morphogenesis, optical, electron and atomic force microscopy has been extensively used to reveal the frustule morphology. However, since silica frustules are opaque, past observations were limited to outer and fracture surfaces, restricting observations of interior structures. Here we show that opaque silica frustules can be converted into electronically transparent graphene replicas, fabricated using chemical vapor deposition of methane. Chemical vapor deposition creates a continuous graphene coating preserving the frustule's shape and fine, complicated internal features. Subsequent dissolution of the silica with hydrofluoric acid yields a free-standing replica of the internal and external native frustule morphologies. Electron microscopy renders these graphene replicas highly transparent, revealing previously unobserved, complex, three-dimensional, interior frustule structures, which lend new insights into the investigation of frustule morphogenesis. PMID:25135739

Study of Cold Coiling Spring Steel on Microstructure and Cold Forming Performance

NASA Astrophysics Data System (ADS)

Jiang, Y.; Liang, Y. L.; Ming, Y.; Zhao, F.

2017-09-01

Medium-carbon cold-coiling locomotive spring steels were treated by a novel Q-P-T (quenching-partitioning-tempering) process. Scanning electron microscopy (SEM), transmission electron microscope (TEM) and X-ray diffraction (XRD) were used to characterize the relevant parameters of the steel. Results show that the microstructure of tested steel treated by Q-P-T process is a complex microstructures composed of martensite, bainite and retained austenite. The volume fraction of retained austenite (wt.%) is up to 31%. After pre-deforming and tempering again at 310°C, the plasticity of samples treated by Q-P-T process is still well. Fracture images show that the Q-P-T samples are ductile fracture. It is attributed to the higher volume fraction of the retained austenite and the interactions between the multi-phases in Q-P-T processed sample.

Microstructure and hydrogen induced failure mechanisms in iron-nickel weldments

NASA Astrophysics Data System (ADS)

Fenske, Jamey Alan

A recent series of inexplicable catastrophic failures of specific subsea dissimilar metal Fe-Ni butter welds has illuminated a fundamental lack of understanding of both the microstructure created along the fusion line as well as its impact on the hydrogen susceptibility of these interfaces. In order to remedy this, the present work compares and contrasts the microstructure and hydrogen-induced fracture morphology of AISI 8630-IN 625 and F22-IN 625 dissimilar metal weld interfaces as a function of post-weld heat treatment duration. A variety of techniques were used to study details of both the microstructure and fracture morphology including optical microscopy, scanning electron microscopy, secondary ion mass spectrometry, transmission electron microscopy, electron backscatter diffraction, and energy dispersive x-ray spectroscopy. For both systems, the microstructure along the weld interface consisted of a coarse grain heat-affected zone in the Fe-base metal followed by discontinuous martensitic partially-mixed zones and a continuous partially-mixed zone on the Ni-side of the fusion line. Within the partially mixed zone on the Ni-side there exists a 200 nm-wide transition zone within a 20 mum-wide planar solidification region followed by a cellular dendritic region with Nb-Mo rich carbides decorating the dendrite boundaries. The size, area fraction and composition of the discontinuous PMZ were determined to be controlled by uneven mixing in the liquid weld pool influenced by convection currents produced from the welding procedure. The virgin martensitic microstructure produced in these regions is formed as consequence of a both the local composition and the post-weld heat treatment. The local higher Ni content results in these regions being retransformed into austenite during the post-weld heat treatment and then virgin martensite while cooling to room temperature. Although there were differences in the volume of the discontinuous partially mixed-zones, the major difference in the weld metal interfaces was the presence of M 7C3 precipitates in the planar solidification region. The formation of these precipitates, which were found in what was previously referred to as the "featureless-zone," were determined to be dependent on the carbon content of the Fe-base metal and the duration of the post-weld heat treatment. A high density of these ordered 100 nm-long by 10 nm-wide needle-like precipitates were found in the AISI 8630-IN 625 weldment in the 10 hour post-weld heat treatment condition while only the initial stages of their nucleation were evident in the F22-IN 625 15 hour post-weld heat treatment specimen. The study of the fractured specimens revealed that the M7C 3 carbides play a key role in the susceptibility to hydrogen embrittlement of the Fe-Ni butter weldments. The fractures initially nucleate along the isolated Fe-base metal -- discontinuous partially mixed zone interfaces. The M7C3 carbides accumulate hydrogen and then provide a low energy fracture path between the discontinuous partially mixed zones leading to catastrophic failure. The result is a fracture morphology that alternates between flat regions produced by fracture along the discontinuous partially mixed zones and cleavage-like fracture regions produced by fracture along the ordered carbide matrix interfaces.

The fracture properties and mechanical design of human fingernails.

PubMed

Farren, L; Shayler, S; Ennos, A R

2004-02-01

Fingernails are a characteristic feature of primates, and are composed of three layers of the fibrous composite keratin. This study examined the structure and fracture properties of human fingernails to determine how they resist bending forces while preventing fractures running longitudinally into the nail bed. Nail clippings were first torn manually to examine the preferred crack direction. Next, scissor cutting tests were carried out to compare the fracture toughness of central and outer areas in both the transverse and longitudinal direction. The fracture toughness of each of the three isolated layers was also measured in this way to determine their relative contributions to the toughness. Finally, the structure was examined by carrying out scanning electron microscopy of free fracture surfaces and polarized light microscopy of nail sections. When nails were torn, cracks were always diverted transversely, parallel to the free edge of the nail. Cutting tests showed that this occurred because the energy to cut nails transversely, at approximately 3 kJ m(-2), was about half that needed (approx. 6 kJ m(-2)) to cut them longitudinally. This anisotropy was imparted by the thick intermediate layer, which comprises long, narrow cells that are oriented transversely; the energy needed to cut this layer transversely was only a quarter of that needed to cut it longitudinally. In contrast the tile-like cells in the thinner dorsal and ventral layers showed isotropic behaviour. They probably act to increase the nail's bending strength, and as they wrap around the edge of the nail, they also help prevent cracks from forming. These results cast light on the mechanical behaviour and care of fingernails.

Fracture toughness improvements of dental ceramic through use of yttria-stabilized zirconia (YSZ) thin-film coatings.

PubMed

Chan, Ryan N; Stoner, Brian R; Thompson, Jeffrey Y; Scattergood, Ronald O; Piascik, Jeffrey R

2013-08-01

The aim of this study was to evaluate strengthening mechanisms of yttria-stabilized zirconia (YSZ) thin film coatings as a viable method for improving fracture toughness of all-ceramic dental restorations. Bars (2mm×2mm×15mm, n=12) were cut from porcelain (ProCAD, Ivoclar-Vivadent) blocks and wet-polished through 1200-grit using SiC abrasive. A Vickers indenter was used to induce flaws with controlled size and geometry. Depositions were performed via radio frequency magnetron sputtering (5mT, 25°C, 30:1 Ar/O2 gas ratio) with varying powers of substrate bias. Film and flaw properties were characterized by optical microscopy, scanning electron microscopy (SEM), and X-ray diffraction (XRD). Flexural strength was determined by three-point bending. Fracture toughness values were calculated from flaw size and fracture strength. Data show improvements in fracture strength of up to 57% over unmodified specimens. XRD analysis shows that films deposited with higher substrate bias displayed a high %monoclinic volume fraction (19%) compared to non-biased deposited films (87%), and resulted in increased film stresses and modified YSZ microstructures. SEM analysis shows critical flaw sizes of 67±1μm leading to fracture toughness improvements of 55% over unmodified specimens. Data support surface modification of dental ceramics with YSZ thin film coatings to improve fracture toughness. Increase in construct strength was attributed to increase in compressive film stresses and modified YSZ thin film microstructures. It is believed that this surface modification may lead to significant improvements and overall reliability of all-ceramic dental restorations. Copyright © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

Buckling of C60 whiskers

NASA Astrophysics Data System (ADS)

Asaka, Koji; Kato, Ryoei; Miyazawa, Kun'ichi; Kizuka, Tokushi

2006-08-01

The authors demonstrated the mechanics of materials for crystalline whiskers composed of C60 molecules; compressive deformation of the whiskers was observed by in situ transmission electron microscopy with simultaneous force measurement by means of an optical cantilever method, as used in atomic force microscopy. In response to compression along the long axis, the whiskers bent first elastically, then buckled. A whisker with 160nm diameter fractured brittlely at a strain of 0.08. According to Euler's formula, Young's modulus of the whisker was estimated to be 32-54GPa, which is 160%-650% of that of C60 bulk crystals.

Thylakoid membrane landscape in the sixties: a tribute to Andrew Benson.

PubMed

Anderson, Jan M

2007-05-01

Prior to the 1960s, the model for the molecular structure of cell membranes consisted of a lipid bilayer held in place by a thin film of electrostatically-associated protein stretched over the bilayer surface: (the Danielli-Davson-Robertson "unit membrane" model). Andrew Benson, an expert in the lipids of chloroplast thylakoid membranes, questioned the relevance of the unit membrane model for biological membranes, especially for thylakoid membranes, instead of emphasizing evidence in favour of hydrophobic interactions of membrane lipids within complementary hydrophobic regions of membrane-spanning proteins. With Elliot Weier, Benson postulated a remarkable subunit lipoprotein monolayer model for thylakoids. Following the advent of freeze fracture microscopy and the fluid lipid-protein mosaic model by Singer and Nicolson, the subunits, membrane-spanning integral proteins, span a dynamic lipid bilayer. Now that high resolution X-ray structures of photosystems I and II are being revealed, the seminal contribution of Andrew Benson can be appreciated.

Morphological and chemical information in fresh and vitrified ovarian tissues revealed by X-ray Microscopy and Fluorescence: observational study

NASA Astrophysics Data System (ADS)

Pascolo, L.; Venturin, I.; Gianoncelli, A.; Salomé, M.; Altissimo, M.; Bedolla, D. E.; Giolo, E.; Martinelli, M.; Luppi, S.; Romano, F.; Zweyer, M.; Ricci, G.

2018-06-01

Many clinical circumstances impose the necessity of collection and prolonged storage of gametes and/or ovarian tissue in order to preserve the reproduction potential of subjects. This is particularly appropriate in the case of young women and pre-pubertal girls undergoing chemotherapeutic treatments. The success of later assisted fertilization will depend on the suitable cooling protocols minimizing cryo-damages and preserving their biological function. The freeze-thaw processes of cryopreservation may induce, in fact, morphological and structural damages of oocytes and tissue mainly due to the formation of intracellular ice and to the toxicity of cryoprotectant. The most used cryo-protocol is the slow freezing procedure, but recently many authors have proposed vitrification as an alternative, because of its simplicity. The damage extent and the quality of follicles after cryopreservation are usually evaluated morphologically by conventional histological procedures, light and electron microscopy. Our laboratory, to further improve the evaluation and to better investigate damages, is adopting a combination of Synchrotron soft X-ray Microscopy (at TwinMic – Elettra) and XRF at different incident energies (at TwinMic – Elettra and ID21 – ESRF). X-ray techniques were performed on histological sections at micro and sub-micron resolution. Phase contrast and absorption images revealed changes in the compactness of the tissues, as well as cellular abnormalities revealed at sub-micrometric resolution. The distributions of the elements detected at 7.3 and 1.5 keV were compared and particularly Cl resulted to be indicative of follicle integrity. The results demonstrate the utility and the potential of X-ray microscopy and fluorescence in this research field.

Distinguishing magnetic blocking and surface spin-glass freezing in nickel ferrite nanoparticles

NASA Astrophysics Data System (ADS)

Nadeem, K.; Krenn, H.; Traussing, T.; Letofsky-Papst, I.

2011-01-01

Nickel ferrite nanoparticles dispersed in SiO2 matrix have been synthesized by sol-gel method. Structural analysis has been performed by using x-ray diffraction and transmission electron microscopy. Magnetic properties have been investigated by using superconducting quantum interference device magnetometry. In addition to the average blocking temperature peak at TB=120 K measured by a zero field cooled temperature scan of the dc susceptibility, an additional hump near 15 K is observed. Temperature dependent out-of-phase ac susceptibility shows the same features: one broad peak at high temperature and a second narrow peak at low temperature. The high temperature peak corresponds to magnetic blocking of individual nanoparticles, while the low temperature peak is attributed to surface spin-glass freezing which becomes dominant for decreasing particle diameter. To prove the dynamics of the spin (dis)order in both regimes of freezing and blocking, the frequency dependent ac susceptibility is investigated under a biasing dc field. The frequency shift in the "frozen" low-temperature ac susceptibility peak is fitted to a dynamic scaling law with a critical exponent zv=7.5, which indicates a spin-glass phase. Exchange bias is turned on at low temperature which signifies the existence of a strong core-shell interaction. Aging and memory effects are further unique fingerprints of a spin-glass freezing on the surface of isolated magnetic nanoparticles.

Spray-Freeze Drying: a Suitable Method for Aerosol Delivery of Antibodies in the Presence of Trehalose and Cyclodextrins.

PubMed

Pouya, Maryam Amini; Daneshmand, Behnaz; Aghababaie, Shabnam; Faghihi, Homa; Vatanara, Alireza

2018-05-08

We aimed to prepare spray-freeze-dried powder of IgG considering physicochemical stability and aerodynamic aspects. Spray-freeze drying (SFD) exposes proteins to various stresses which should be compensated by suitable stabilizers. The competence of cyclodextrins (CDs), namely beta-cyclodextrin (βCD) and hydroxypropyl βCD (HPβCD), at very low concentrations, was investigated in the presence of separate mannitol- and trehalose-based formulations. Spray-freeze-dried preparations were quantified in terms of monomer recovery and conformation by size exclusion chromatography (SEC-HPLC) and Fourier transform infrared (FTIR) spectroscopy, respectively. Differential scanning calorimetry (DSC) and X-ray diffractometry (XRD) were employed to identify the thermal characteristics of powders. Particle morphology was visualized by scanning electron microscopy (SEM). Aerodynamic behavior of powders was checked through an Anderson cascade impactor (ACI). Although all formulations protected antibody from aggregation during the SFD process (aggregation < 1%), mannitol-containing ones failed upon the storage (19.54% in the worst case). Trehalose-HPβCD incomparably preserved the formulation with fine particle fraction (FPF) of 51.29%. Crystallization of mannitol resulted in IgG destabilization upon storage. Although employed concentration of CDs is too low (less than 50:1 molar ratio to protein), they successfully served as stabilizing agents in SFD with perfect improvement in aerosol functionality. Graphical Abstract ᅟ.

Doxycycline hydrochloride-metronidazole solid lipid microparticles gels for treatment of periodontitis: development, in-vitro and in-vivo clinical evaluation.

PubMed

Gad, Heba A; Kamel, Amany O; Ezzat, Ola M; El Dessouky, Hadir F; Sammour, Omaima A

2017-11-01

To formulate solid lipid microparticles (SLMs) encapsulating doxycycline hydrochloride (DH) and metronidazole (MT) for the treatment of periodontal diseases. SLMs were prepared applying hot homogenization method, using different types of lipids and stabilized with various types and concentrations of surfactants. The optimized formula was subjected to freeze-drying followed by incorporation into poloxamer gel. Microbiological and clinical evaluation of the selected SLMs on patients suffering from periodontal diseases was performed. SLMs could entrap high percentage of both drugs (81.14% and 68.75 % for doxycycline hydrochloride and metronidazole respectively). Transmission electron microscopy images of SLMs showed nearly spherical particles. Freeze-dried SLMs showed satisfactory stability for three months. Combined drugs were molecularly dispersed in SLMs. Incorporation of the freeze-dried SLMs powder in poloxamer gel could control the drugs release for 72 h. In-vivo study revealed effective and safe use of SLMs gel for periodontitis treatment. Significant improvement in both microbiological and clinical parameters was observed as compared to scaling and root planing alone. The formulated SLMs gel offers an applicable dosage form that can be injected directly into the periodontal pocket as adjunctive to scaling and root planing.

Ice Nucleation Activity of Graphene and Graphene Oxides

PubMed Central

2018-01-01

Aerosols can act as cloud condensation nuclei and/or ice-nucleating particles (INPs), influencing cloud properties. In particular, INPs show a variety of different and complex mechanisms when interacting with water during the freezing process. To gain a fundamental understanding of the heterogeneous freezing mechanisms, studies with proxies for atmospheric INPs must be performed. Graphene and its derivatives offer suitable model systems for soot particles, which are ubiquitous aerosols in the atmosphere. In this work, we present an investigation of the ice nucleation activity (INA) of different types of graphene and graphene oxides. Immersion droplet freezing experiments as well as additional analytical analyses, such as X-ray photoelectron spectroscopy, Raman spectroscopy, and transmission electron microscopy, were performed. We show within a group of samples that a highly ordered graphene lattice (Raman G band intensity >50%) can support ice nucleation more effectively than a lowly ordered graphene lattice (Raman G band intensity <20%). Ammonia-functionalized graphene revealed the highest INA of all samples. Atmospheric ammonia is known to play a primary role in the formation of secondary particulate matter, forming ammonium-containing aerosols. The influence of functionalization on interactions between the particle interface and water molecules, as well as on hydrophobicity and agglomeration processes, is discussed. PMID:29707097

Micromechanisms of brittle fracture: STM, TEM and electron channeling analysis. Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gerberich, W.W.

1997-01-01

The original thrust of this grant was to apply newly developed techniques in scanning tunneling and transmission electron microscopy to elucidate the mechanism of brittle fracture. This grant spun-off several new directions in that some of the findings on bulk structural materials could be utilized on thin films or intermetallic single crystals. Modeling and material evaluation efforts in this grant are represented in a figure. Out of this grant evolved the field the author has designated as Contact Fracture Mechanics. By appropriate modeling of stress and strain distribution fields around normal indentations or scratch tracks, various measures of thin filmmore » fracture or decohesion and brittle fracture of low ductility intermetallics is possible. These measures of fracture resistance in small volumes are still evolving and as such no standard technique or analysis has been uniformly accepted. For brittle ceramics and ceramic films, there are a number of acceptable analyses such as those published by Lawn, Evans and Hutchinson. For more dissipative systems involving metallic or polymeric films and/or substrates, there is still much to be accomplished as can be surmised from some of the findings in the present grant. In Section 2 the author reviews the funding history and accomplishments associated mostly with bulk brittle fracture. This is followed by Section 3 which covers more recent work on using novel techniques to evaluate fracture in low ductility single crystals or thin films using micromechanical probes. Basically Section 3 outlines how the recent work fits in with the goals of defining contact fracture mechanics and gives an overview of how the several examples in Section 4 (the Appendices) fit into this framework.« less

Delivery of femtolitre droplets using surface acoustic wave based atomisation for cryo-EM grid preparation.

PubMed

Ashtiani, Dariush; Venugopal, Hari; Belousoff, Matthew; Spicer, Bradley; Mak, Johnson; Neild, Adrian; de Marco, Alex

2018-04-06

Cryo-Electron Microscopy (cryo-EM) has become an invaluable tool for structural biology. Over the past decade, the advent of direct electron detectors and automated data acquisition has established cryo-EM as a central method in structural biology. However, challenges remain in the reliable and efficient preparation of samples in a manner which is compatible with high time resolution. The delivery of sample onto the grid is recognized as a critical step in the workflow as it is a source of variability and loss of material due to the blotting which is usually required. Here, we present a method for sample delivery and plunge freezing based on the use of Surface Acoustic Waves to deploy 6-8 µm droplets to the EM grid. This method minimises the sample dead volume and ensures vitrification within 52.6 ms from the moment the sample leaves the microfluidics chip. We demonstrate a working protocol to minimize the atomised volume and apply it to plunge freeze three different samples and provide proof that no damage occurs due to the interaction between the sample and the acoustic waves. Copyright © 2018 Elsevier Inc. All rights reserved.

Influent of Borax Decahydrate Composition as Additional Flux into Stoneware Bodies

NASA Astrophysics Data System (ADS)

Bakil, Siti Natrah Abd; Hussin, Rosniza; Bakar Aramjat, Abu

2017-08-01

Stoneware is vitrified, has less porosity and requires high sintering temperature. The influent of borax decahydrate composition at sintering temperature 1050°C and 1150°C on the thermal analysis, fracture surface, linear shrinkage, water absorption and modular of rapture (MOR) were investigated. Rectangular sample were produced by uniaxially pressing at 40MPa. The thermal behavior was determined by thermogravimetric and different thermal analysis (TGA-DTA). The Scanning electron microscopy (SEM) was used for fracture surface analysis. The water absorption (%) of the sample were determined using Archimedes’ method. The experimental result showed that content of borax decahydrate have influent the properties of stoneware bodies.

Thermal-stress fracture and fractography in UO/sub 2/

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kennedy, C.R.; Bandyopadhyay, G.

1976-01-01

Pressed and sintered UO/sub 2/ pellets were thermally shocked by quenching into a water bath at room temperature. The cracking behavior and strength degradation, as measured by the diametral compression technique, in these quench tests are discussed. Fractography of the thermally shocked specimens by scanning-electron microscopy indicated predominantly intergranular fracture in UO/sub 2/ in severe thermal-shock tests. The implication of this observation is that intergranular cracking may occur during the initial heat up in a reactor. Because fission gas bubbles tend to migrate toward the grain boundary, preferential microcracking along the boundary may strongly affect subsequent fission gas release behavior.

Functional Reconstitution into Liposomes of Purified Human RhCG Ammonia Channel

PubMed Central

Mouro-Chanteloup, Isabelle; Cochet, Sylvie; Chami, Mohamed; Genetet, Sandrine; Zidi-Yahiaoui, Nedjma; Engel, Andreas; Colin, Yves; Bertrand, Olivier; Ripoche, Pierre

2010-01-01

Background Rh glycoproteins (RhAG, RhBG, RhCG) are members of the Amt/Mep/Rh family which facilitate movement of ammonium across plasma membranes. Changes in ammonium transport activity following expression of Rh glycoproteins have been described in different heterologous systems such as yeasts, oocytes and eukaryotic cell lines. However, in these complex systems, a potential contribution of endogenous proteins to this function cannot be excluded. To demonstrate that Rh glycoproteins by themselves transport NH3, human RhCG was purified to homogeneity and reconstituted into liposomes, giving new insights into its channel functional properties. Methodology/Principal Findings An HA-tag introduced in the second extracellular loop of RhCG was used to purify to homogeneity the HA-tagged RhCG glycoprotein from detergent-solubilized recombinant HEK293E cells. Electron microscopy analysis of negatively stained purified RhCG-HA revealed, after image processing, homogeneous particles of 9 nm diameter with a trimeric protein structure. Reconstitution was performed with sphingomyelin, phosphatidylcholine and phosphatidic acid lipids in the presence of the C12E8 detergent which was subsequently removed by Biobeads. Control of protein incorporation was carried out by freeze-fracture electron microscopy. Particle density in liposomes was a function of the Lipid/Protein ratio. When compared to empty liposomes, ammonium permeability was increased two and three fold in RhCG-proteoliposomes, depending on the Lipid/Protein ratio (1/300 and 1/150, respectively). This strong NH3 transport was reversibly inhibited by mercuric and copper salts and exhibited a low Arrhenius activation energy. Conclusions/Significance This study allowed the determination of ammonia permeability per RhCG monomer, showing that the apparent PunitNH3 (around 1×10−3 µm3.s−1) is close to the permeability measured in HEK293E cells expressing a recombinant human RhCG (1.60×10−3 µm3.s−1), and in human red blood cells endogenously expressing RhAG (2.18×10−3 µm3.s−1). The major finding of this study is that RhCG protein is active as an NH3 channel and that this function does not require any protein partner. PMID:20126667

[Scanning electron microscopy of heat-damaged bone tissue].

PubMed

Harsanyl, L

1977-02-01

Parts of diaphyses of bones were exposed to high temperature of 200-1300 degrees C. Damage to the bone tissue caused by the heat was investigated. The scanning electron microscopic picture seems to be characteristic of the temperature applied. When the bones heated to the high temperature of 700 degrees C characteristic changes appear on the periostal surface, higher temperatura on the other hand causes damage to the compact bone tissue and can be observed on the fracture-surface. Author stresses the importance of this technique in the legal medicine and anthropology.

Multiphase materials with lignin. VI. Effect of cellulose derivative structure on blend morphology with lignin

Treesearch

Timothy G. Rials; Wolfgang G. Glasser

1989-01-01

Polymeric blends of lignin with ethyl cellulose (EC) and cellulose acetate/butyrate (CAB) prepared by solution casting from dioxane. Fracture surface analysis by scanning electron microscopy revealed phase separation when the lignin content exceeded 10% for blends with EC and 5% in the CAB system. While this phase behavior is as predicted for the EC blends, a greater...

An investigation of the constituent elements and modes of fracture of in vivo fractured metallic posts.

PubMed

Fox, Kathryn; Wood, David J; Youngson, Callum C

2007-01-01

The objective of this investigation was to determine the materials used in metallic post and core construction and investigate and the modes of failure that occurred resulting in post fracture. Eighty-five fractured metal posts were clinically retrieved over a 5-year period. The tooth with the fractured post, and relevant clinical details were recorded. Investigation of the fractured posts was undertaken to determine their constituent elements and modes of fracture using scanning electron microscopy (SEM) and energy-dispersive X-ray analysis (EDX). The results showed that 77% of the posts were cast and 23% wrought. The modes of fracture were ductile (67%) or brittle (52%) although some posts exhibited both within the same specimen. In addition, evidence of fatigue was present in 20% of posts and porosity in 48%. A large variety of materials were used in the construction of post and cores although low-gold silver-palladium alloy most commonly encountered. Their failure was predominantly ductile in nature although a significant amount of brittle fracture was also noted. Porosity within castings also appeared to be a contributory factor in some cases and this was particularly evident in high-palladium castings. Evidence of fatigue was noted in a minority of posts.

Effect of Low-Temperature Environment on Stress Corrosion Cracking Behavior of X80 Pipeline Steel in Simulated Alkaline Soil Solution

NASA Astrophysics Data System (ADS)

Xie, Fei; Wang, Dan; Wu, Ming; Yu, Chengxiang; Sun, Dongxu; Yang, Xu; Xu, Changhao

2018-04-01

The stress corrosion cracking (SCC) of X80 pipeline steel in simulated alkaline soil solution under different temperatures was investigated by slow-strain-rate testing, scanning electron microscopy and energy-dispersive spectroscopy. Results showed that the fracture was transgranular and brittle at 273 K to 278 K (0 °C to 5 °C), and the metal surface was dissolved by a large number of chloride ions. Furthermore, hydrogen embrittlement was caused by the hydrogen atom extended to the high-stress region. The fracture process was controlled by hydrogen-induced cracking, and SCC was highly sensitive at this stage. At 288 K to 298 K (15 °C to 25 °C), the fracture morphology was attributed to the mixed mode of ductile and brittle fractures, the fracture process was controlled by the mechanism of hydrogen-induced cracking and anodic dissolution, and the susceptibility to SCC decreased. When the temperature reached 308 K to 318 K (35 °C to 45 °C), the fracture was mainly intergranular and ductile, the fracture process was controlled by anodic dissolution, and SCC sensitivity was the smallest in this temperature range.

Scanning electron microscopic observations of 'fractured' biodegradable plates and screws.

PubMed

Kosaka, Masaaki; Uemura, Fumiko; Tomemori, Shoko; Kamiishi, Hiroshi

2003-02-01

We encountered two out of 100 cases in which implanted biodegradable plates and screws had fractured within 1 month postoperatively. Failure of the material was confirmed through clinical symptoms, radiographs or CT findings. In addition, four specimens obtained from these two cases were examined with regard to their ultrastructure using scanning electron microscopy. Several principal patterns of the fractured surface were found: (1) gradual cracking, i.e. 'circular stair' and, (2) tortuous threads, i.e. a wavy line. It is conceivable that the material may not have been hit by major sudden forces but a disproportion between the thread configuration and the drilled hole may have led to screw loosening and torsion. Subsequently, the threads were deformed in a 'wavy' manner, finally leading to cracking and fracture of plates and screws. Fractures of plates and screws due to these instabilities are thought to be distinguishable from material resorption. In the application of biodegradable materials, more than two screws per single bone segment should be used as a principle of plate-fixation technique in order to avoid a stability-compromising situation, particularly in the stress-bearing areas of the maxillofacial region. Moreover, three-dimensional fixation using more than two plates is recommended in the facial skeleton e.g. zygomatic tripod. Intermaxillary fixation should also be considered to reinforce initial stability in stress-bearing areas.

A study of hydrogen environment effects on microstructure property behavior of NASA-23 alloy and related alloy systems

NASA Technical Reports Server (NTRS)

Diwan, Ravinder M.

1990-01-01

This work is part of the overall advanced main combustion chamber (AMCC) casting characterization program of the Materials and Processes Laboratory of the Marshall Space Flight Center. The influence of hydrogen on the tensile properties and ductility behavior of NASA-23 alloy were analyzed. NASA-23 and other referenced alloys in cast and hipped conditions were solution treated and aged under selected conditions and characterized using optical metallography, scanning electron microscopy, and electron microprobe analysis techniques. The yield strength of NASA-23 is not affected much by hydrogen under tensile tests carried at 5000 psig conditions; however, the ultimate strength and ductility properties are degraded. This implies that the physical mechanisms operating would be related to the plastic deformation process. The fracture surfaces characteristics of NASA-23 specimens tensile tested in hydrogen, helium, and air were also analyzed. These revealed surface cracks around specimen periphery with the fracture surface showing a combination of intergranular and transgranular modes of fracture. It is seen that the specimens charged in hydrogen seem to favor a more brittle fracture mode in comparison to air and helium charged specimens. The AMCC casting characterization program is to be analyzed for their hydrogen behavior. As a result of this program, the basic microstructural factors and fracture characteristics in some cases were analyzed.

Detachment of particulate iron sulfide during shale-water interaction

NASA Astrophysics Data System (ADS)

Emmanuel, S.; Kreisserman, Y.

2017-12-01

Hydraulic fracturing, a commonly used technique to extract oil and gas from shales, is controversial in part because of the threat it poses to water resources. The technique involves the injection into the subsurface of large amounts of fluid, which can become contaminated by fluid-rock interaction. The dissolution of pyrite is thought to be a primary pathway for the contamination of fracturing fluids with toxic elements, such as arsenic and lead. In this study, we use direct observations with atomic force microscopy to show that the dissolution of carbonate minerals in Eagle Ford shale leads to the physical detachment of embedded pyrite grains. To simulate the way fluid interacts with a fractured shale surface, we also reacted rock samples in a flow-through cell, and used environmental scanning electron microscopy to compare the surfaces before and after interaction with water. Crucially, our results show that the flux of particulate iron sulfide into the fluid may be orders of magnitude higher than the flux of pyrite from chemical dissolution. This result suggests that mechanical detachment of pyrite grains could be the dominant mode by which arsenic and other inorganic elements are mobilized in the subsurface. Thus, during hydraulic fracturing operations and in groundwater systems containing pyrite, the transport of many toxic species may be controlled by the transport of colloidal iron sulfide particles.

The response of single human cells to zero gravity

NASA Technical Reports Server (NTRS)

Montgomery, P. O., Jr.; Cook, J. E.; Reynolds, R. C.; Paul, J. S.; Hayflick, L.; Stock, D.; Schulz, W. W.; Kimzey, S. L.; Thirolf, R. G.; Rogers, T.

1974-01-01

The SO15 experiment was designed to extend observations of the effects of zero-gravity to living human cells during and subsequent to a 59-day flight on Skylab 3. A strain of diploid human embryonic lung cells, WI-38, was chosen for this purpose. The studies were concerned with observations designed to detect the effects of zero-gravity on cell growth rates and on cell structure as observed by light microscopy, transmission and scanning electron microscopy and histochemistry. Studies of the effects of zero-gravity on the cell function and the cell cycle were performed by time lapse motion picture photography and microspectrophotometry. Subsequent study of the returned living cells included karotyping, G- and C-banding, and analyses of the culture media used. Some of the living cells returned were banked by deep freeze techniques for possible future experiments.

Scanning electron microscopy of the vestibular end organs. [morphological indexes of inner ear anatomy and microstructure

NASA Technical Reports Server (NTRS)

Lindeman, H. H.; Ades, H. W.; West, R. W.

1973-01-01

The vestibular end organs, after chemical fixation, were freeze dried, coated with gold and palladium, and studied in the scanning microscope. Scanning microscopy gives a good three dimensional view of the sensory areas and allows study of both gross anatomy and microstructures. Cross anatomical features of the structure of the ampullae are demonstrated. The form of the statoconia in different species of animals is shown. New aspects of the structure of the sensory hairs are revealed. The hair bundles in the central areas of the cristae and in the striola of the maculae differ structurally from the hair bundles at the periphery of the sensory regions. Furthermore, some hair bundles consisting of very short stereocilia were observed. The relationship between the cupula and the statoconial membrane to the epithelial surface is discussed.

Freeze-fracture study of the filamentous, segmented microorganism attached to the murine small bowel.

PubMed Central

Snellen, J E; Savage, D C

1978-01-01

A freeze-fracture study has provided new information about the filamentous, segmented microorganism known to live in the murine small bowel. The intracellular bodies produced by this microbe appear to arise by a modified sporogenesis so that they are enclosed in an envelopment membrane at least prior to release by the filament mother cell. At least some of the intracellular bodies divide while still within the mother cell, suggesting a reproductive role for these structures. The host epithelial membrane remains intact at the site of attachment, but does appear to have a reduced concentration of intramembrane particles. Changes in the host cytoplasm adjacent to the attachment site are documented and interpreted to be a sol-gel transformation which may stabilize the attachment socket. Images PMID:659364

Lithium-assisted electrochemical welding in silicon nanowire battery electrodes.

PubMed

Karki, Khim; Epstein, Eric; Cho, Jeong-Hyun; Jia, Zheng; Li, Teng; Picraux, S Tom; Wang, Chunsheng; Cumings, John

2012-03-14

From in situ transmission electron microscopy (TEM) observations, we present direct evidence of lithium-assisted welding between physically contacted silicon nanowires (SiNWs) induced by electrochemical lithiation and delithiation. This electrochemical weld between two SiNWs demonstrates facile transport of lithium ions and electrons across the interface. From our in situ observations, we estimate the shear strength of the welded region after delithiation to be approximately 200 MPa, indicating that a strong bond is formed at the junction of two SiNWs. This welding phenomenon could help address the issue of capacity fade in nanostructured silicon battery electrodes, which is typically caused by fracture and detachment of active materials from the current collector. The process could provide for more robust battery performance either through self-healing of fractured components that remain in contact or through the formation of a multiconnected network architecture. © 2012 American Chemical Society

Tensile Deformation Temperature Impact on Microstructure and Mechanical Properties of AISI 316LN Austenitic Stainless Steel

NASA Astrophysics Data System (ADS)

Xiong, Yi; He, Tiantian; Lu, Yan; Ren, Fengzhang; Volinsky, Alex A.; Cao, Wei

2018-03-01

Uniaxial tensile tests were conducted on AISI 316LN austenitic stainless steel from - 40 to 300 °C at a rate of 0.5 mm/min. Microstructure and mechanical properties of the deformed steel were investigated by optical, scanning and transmission electron microscopies, x-ray diffraction, and microhardness testing. The yield strength, ultimate tensile strength, elongation, and microhardness increase with the decrease in the test temperature. The tensile fracture morphology has the dimple rupture feature after low-temperature deformations and turns to a mixture of transgranular fracture and dimple fracture after high-temperature ones. The dominating deformation microstructure evolves from dislocation tangle/slip bands to large deformation twins/slip bands with temperature decrease. The deformation-induced martensite transformation can only be realized at low temperature, and its quantity increases with the decrease in the temperature.

Focus on membrane differentiation and membrane domains in the prokaryotic cell.

PubMed

Boekema, Egbert J; Scheffers, Dirk-Jan; van Bezouwen, Laura S; Bolhuis, Henk; Folea, I Mihaela

2013-01-01

A summary is presented of membrane differentiation in the prokaryotic cell, with an emphasis on the organization of proteins in the plasma/cell membrane. Many species belonging to the Eubacteria and Archaea have special membrane domains and/or membrane proliferation, which are vital for different cellular processes. Typical membrane domains are found in bacteria where a specific membrane protein is abundantly expressed. Lipid rafts form another example. Despite the rareness of conventional organelles as found in eukaryotes, some bacteria are known to have an intricate internal cell membrane organization. Membrane proliferation can be divided into curvature and invaginations which can lead to internal compartmentalization. This study discusses some of the clearest examples of bacteria with such domains and internal membranes. The need for membrane specialization is highest among the heterogeneous group of bacteria which harvest light energy, such as photosynthetic bacteria and halophilic archaea. Most of the highly specialized membranes and domains, such as the purple membrane, chromatophore and chlorosome, are found in these autotrophic organisms. Otherwise the need for membrane differentiation is lower and variable, except for those structures involved in cell division. Microscopy techniques have given essential insight into bacterial membrane morphology. As microscopy will further contribute to the unraveling of membrane organization in the years to come, past and present technology in electron microscopy and light microscopy is discussed. Electron microscopy was the first to unravel bacterial morphology because it can directly visualize membranes with inserted proteins, which no other technique can do. Electron microscopy techniques developed in the 1950s and perfected in the following decades involve the thin sectioning and freeze fractioning of cells. Several studies from the golden age of these techniques show amazing examples of cell membrane morphology. More recently, light microscopy in combination with the use of fluorescent dyes has become an attractive technique for protein localization with the natural membrane. However, the resolution problem in light microscopy remains and overinterpretation of observed phenomena is a pitfall. Thus, light microscopy as a stand-alone technique is not sufficient to prove, for instance, the long-range helical distribution of proteins in membrane such as MinD spirals in Bacillus subtilis. Electron tomography is an emerging electron microscopy technique that can provide three-dimensional reconstructions of small, nonchemically fixed bacteria. It will become a useful tool for studying prokaryotic membranes in more detail and is expected to collect information complementary to those of advanced light microscopy. Together, microscopy techniques can meet the challenge of the coming years: to specify membrane structures in more detail and to bring them to the level of specific protein-protein interactions. Copyright © 2013 S. Karger AG, Basel.

Role of different compatibilizing approaches on the microstructure and mechanical properties of polypropylene/talc composites

NASA Astrophysics Data System (ADS)

Homayounfar, S. Z.; Bagheri, R.

2014-05-01

Since in a highly filled polymer, a major problem arises from non-uniformity of properties due to the poor dispersion of filler, the application of coupling agents have been directed to overcome this problem and also to enhance the mechanical performance of the composites by improving the adhesion at the interface. In this study, a comparison between two major coupling approaches is conducted: 1) Using PPgMA as a kind of compatibilizer which changes the nature of the matrix, 2) Using titanate coupling agent which takes action at the interface and reacts with hydroxyl groups at the inorganic filler surface, resulting in the formation of monomolecular layer on the inorganic surface to increase compatibility of filler/matrix interface. The comparison is made based on the mechanical properties of the composites by means of elastic modulus, yield stress, impact strength and percentage of strain-to-fracture and evaluation of their effects on both the dispersion and adhesion of talc plates in the matrix through the microscopy. Transmission optical microscopy (TOM) and scanning electron microscopy (SEM) are used to observe the deformation micromechanism and the fracture surface of the composites, respectively.

Scanning electron acoustic microscopy of indentation-induced cracks and residual stresses in ceramics

NASA Technical Reports Server (NTRS)

Cantrell, John H.; Qian, Menglu; Ravichandran, M. V.; Knowles, K. M.

1990-01-01

The ability of scanning electron acoustic microscopy (SEAM) to characterize ceramic materials is assessed. SEAM images of Vickers indentations in SiC whisker-reinforced alumina clearly reveal not only the radial cracks, the length of which can be used to estimate the fracture toughness of the material, but also reveal strong contrast, interpreted as arising from the combined effects of lateral cracks and the residual stress field left in the SiC whisker-reinforced alumina by the indenter. The strong contrast is removed after the material is heat treated at 1000 C to relieve the residual stresses around the indentations. A comparison of these observations with SEAM and reflected polarized light observations of Vickers indentations in soda-lime glass both before and after heat treatment confirms the interpretation of the strong contrast.

Fracture Behaviors of Sn-Cu Intermetallic Compound Layer in Ball Grid Array Induced by Thermal Shock

NASA Astrophysics Data System (ADS)

Shen, Jun; Zhai, Dajun; Cao, Zhongming; Zhao, Mali; Pu, Yayun

2014-02-01

In this work, thermal shock reliability testing and finite-element analysis (FEA) of solder joints between ball grid array components and printed circuit boards with Cu pads were used to investigate the failure mechanism of solder interconnections. The morphologies, composition, and thickness of Sn-Cu intermetallic compounds (IMC) at the interface of Sn-3.0Ag-0.5Cu lead-free solder alloy and Cu substrates were investigated by scanning electron microscopy and transmission electron microscopy. Based on the experimental observations and FEA results, it can be recognized that the origin and propagation of cracks are caused primarily by the difference between the coefficient of thermal expansion of different parts of the packaged products, the growth behaviors and roughness of the IMC layer, and the grain size of the solder balls.

Particle shape effect on erosion of optical glass substrates due to microparticles

NASA Astrophysics Data System (ADS)

Waxman, Rachel; Gray, Perry; Guven, Ibrahim

2018-03-01

Impact experiments using sand particles and soda lime glass spheres were performed on four distinct glass substrates. Sand particles were characterized using optical and scanning electron microscopy. High-speed video footage from impact tests was used to calculate incoming and rebound velocities of the individual impact events, as well as the particle volume and two-dimensional sphericity. Furthermore, video analysis was used in conjunction with optical and scanning electron microscopy to relate the incoming velocity and particle shape to subsequent fractures, including both radial and lateral cracks. Indentation theory [Marshall et al., J. Am. Ceram. Soc. 65, 561-566 (1982)] was applied and correlated with lateral crack lengths. Multi-variable power law regression was performed, incorporating the particle shape into the model and was shown to have better fit to damage data than the previous indentation model.

Preparation of alpha-alumina-supported mesoporous bentonite membranes for reverse osmosis desalination of aqueous solutions.

PubMed

Li, Liangxiong; Dong, Junhang; Lee, Robert

2004-05-15

In this study, mesoporous bentonite clay membranes approximately 2 microm thick were prepared on porous alpha-alumina substrates by a sol-gel method. Nanosized clay particles were obtained from commercial Na-bentonite powders (Wyoming) by a process of sedimentation, washing, and freeze-drying. X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and nitrogen adsorption-desorption were employed for membrane characterization. It was found that the content of solids, concentration of polymer binder, and pH value of the clay colloidal suspension had critical influences on membrane formation during the dip-coating process. The membranes were tested for reverse osmosis separation of a 0.1 M NaCl solution. Both water permeability and Na(+) rejection rate of the supported membranes were comparable to those of the compacted thick membranes reported in the literature. However, due to the drastically reduced membrane thickness, water permeance and flux of the supported membranes were significantly higher than those of the compacted thick membranes. It was also observed that the calcination temperature played a critical role in determining structural stability in water and desalination performance of the clay membrane.

Dimeric arrangement and structure of the membrane-bound acetylcholine receptor studied by electron microscopy.

PubMed Central

Zingsheim, H P; Neugebauer, D C; Frank, J; Hänicke, W; Barrantes, F J

1982-01-01

The acetylcholine receptor protein (AChR) from the electric organ of Torpedo marmorata is studied in its membrane-bound form by electron microscopy and single-particle image averaging. About half the molecule protrudes from the membrane surface by approximately 5 nm. The low-resolution 3-D structure of this hydrated portion, including its handedness, can be deduced from averaged axial and lateral projections and from freeze-etched membrane surfaces. In native membrane fragments, a dimeric form of the AChR is observed and the relative orientation of the AChR monomers within the dimer is established. The dimers disappear upon disulfide reduction of the membrane preparations, whereas the average axial projections of the AChR monomer remain unaffected. Since the existence of disulfide bonds linking AChR monomers between their respective delta-subunits is well documented, the approximate position of the delta-subunit within the low-resolution structure of the AChR molecule can be deduced from the structure of the dimers. Images Fig. 1. Fig. 2. Fig. 3. PMID:7188351

Integrating two-photon microscopy and cryo-electron microscopy for studying the interaction of Cafeteria roenbergensis and CroV

NASA Astrophysics Data System (ADS)

Aghvami, Seyedmohammadali

Cafeteria roenbergensis (Cro) is a marine zooplankton; its voracious appetite plays a significant role in regulating bacteria populations. The giant virus that lives within Cro, known as Cafeteria roenbergensis virus (CroV), has an important effect on the mortality of Cro populations. Although viral infections are extremely abundant in oceans, the complete procedure of the infection is still unknown. We study the infection process of Cro by CroV to find out whether the initial contact is through phagocytosis or CroV penetrating the host cell membrane directly. Cro is a moving at speed in the range of 10-100 um/s, therefore, there are many difficulties and challenges for traditional imaging techniques to study this viral-host interaction. We apply two-photon fluorescence microscopy to image this infection process. The image is taken at video rate (30 frame/s), which makes us able to catch the moment of interaction. We are able to image host and virus simultaneously where CroV is stained by SYBR gold dye and Cro is excited through NADH autofluorescence. For further structural biology study, we will obtain atomic level resolution information of infection. After catching the initial moment of infection, we will freeze the sample instantly and image it with cryo-electron microscope .

Fracture behaviour of WC-Co hardmetals with WC partially substituted by titanium carbide

NASA Astrophysics Data System (ADS)

Szutkowska, M.; Boniecki, M.; Cygan, S.; Kalinka, A.; Grilli, M. L.; Balos, S.

2018-03-01

The addition of various amounts of TiC0.9 phase in the range from 5wt.% to 20wt.% substituting WC phase was applied in WC-Co hardmetals with 9.5 wt.% bonding cobalt phase. The hardmetals were consolidated using Hot Isostatic Pressing (HIP) method at temperature of 1573K and pressure of 1500 atm. The plain strain fracture toughness has been determined from 3PB test on a precracking single edge notched beam (SENB) specimen. The indentation fracture toughness with Vickers cracks for comparison was also measured, which changed from 12 to 9.0 MPa·m1/2. The amount of the TiC0.9 phase affected the mechanical and physical properties: Vickers hardness from 12.5 to 14.0 GPa, Young’s modulus from 550 to 460 GPa, density from 13.1 to 9.6 g/cm3, friction coefficient from 0.24 to 0.45, fracture toughness from 16.8 to 11.0 MPa·m1/2. Scanning electron microscopy (SEM), X-ray and electron diffraction phase analysis were used to examine the WC-Co hardmetal with addition of the TiC0.9 phase. For comparison, physical and mechanical properties of the WC-Co hardmetals before modification were tested.

Combining Pre- and Post-Nucleation Trajectories for the Synthesis of High FAU-Content Faujasite Nanocrystals from Organic-Free Sols

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khaleel, Maryam; Xu, Wenqian; Lesch, David A.

The effects of synthesis conditions on the FAU/EMT content and the size of nanocrystals, formed from inorganic aluminosilicate sols, were investigated. High resolution transmission electron microscopy imaging and comparison of experimental X-ray diffraction patterns with simulations demonstrated that all materials made starting from synthesis mixtures in the composition range (1.8-33) SiO2: 1 Al2O3: (2.7-33) Na2O: (41-1000) H2O contain FAU/EMT intergrowths. Compositions with low water content increase the FAU fraction up to 0.8 but the crystal size exceeds 100 nm. Extension of the higher FAU purity to nanocrystals was achieved only by first mixing the sol at high water content compositionsmore » that favor nanocrystal formation and then - after a certain time - lowering by freeze-drying the water to levels favoring the formation of FAU. Cryogenic transmission electron microscopy and small angle X-ray scattering from representative optically clear and colloidally stable precursor sols (aged and crystallized at ambient temperature) reveal the formation of amorphous aggregates before the detection of crystals, in agreement with earlier findings and an existing model for the aggregative growth of the zeolite MFI. The presence of these amorphous aggregates coincides with the aforementioned state of sol that preserves the original trajectory towards nano-crystals after the pronounced reduction of water content by freeze-drying. If water reduction by freeze-drying is applied earlier (before the detection of amorphous aggregates), the sol follows the low water content trajectory towards larger crystals. Despite this memory effect, the sol at this stage is still agnostic towards FAU or EMT formation, the relative content of which is dominantly determined by the final water content. These findings demonstrate that it is possible to combine the effects of pre-and post-nucleation sol composition to steer crystal size and crystal structure, respectively. They confirm precursor nanoparticle evolution, while they emphasize the importance of solution phase composition at both pre- and post-nucleation stages of aggregative crystal growth.« less

Novel nanoparticulate carrier system based on carnauba wax and decyl oleate for the dispersion of inorganic sunscreens in aqueous media.

PubMed

Villalobos-Hernández, J R; Müller-Goymann, C C

2005-05-01

The purpose of this study was to characterize carrier systems for inorganic sunscreens based on a matrix composed of carnauba wax and decyl oleate. Ultraviolet radiation attenuators like barium sulfate, strontium carbonate and titanium dioxide were tested. The lipid matrices were used either as capsules or as accompanying vehicles for the pigments in aqueous dispersions. Manufacturing was performed using high pressure homogenization at 300bar and a temperature of 75 degrees C. To evaluate the effect of the pigments on the crystalline structure of the wax-oil mixture, X-ray diffraction and differential scanning calorimetry were used. Further parameters determined were particle size, polydispersity index, z-potential, viscosity and sun protection factor (SPF). Transmission electron microscopy was also applied for visualization of nanoparticles. The X-ray diffraction patterns and the melting points of the lipid mixtures remained unchanged after the pigments were added. The particle sizes of the encapsulated species ranged from 239 to 749.9nm showing polydispersity values between 0.100 and 0.425. Surface charge measurements comprising values up to -40.8mV denoted the presence of stable dispersions. The formulations could be described as ideal viscous presenting viscosities in a range of 1.40-20.5mPas. Significant increases in SPF up to about 50 were reported after the encapsulation of titanium dioxide. Freeze fracture micrographs confirmed the presence of encapsulated inorganic crystals.

Analyzing phorbol ester effects on gap junctional communication: a dramatic inhibition of assembly

PubMed Central

1994-01-01

The effect of 12-O-tetradeconylphorbol-13-acetate (TPA) on gap junction assembly between Novikoff hepatoma cells was examined. Cells were dissociated with EDTA to single cells and then reaggregated to form new junctions. When TPA (25 nM) was added to the cells at the onset of the 60-min reaggregation, dye transfer was detected at only 0.6% of the cell-cell interfaces compared to 72% for the untreated control and 74% for 4-alpha TPA, an inactive isomer of TPA. Freeze-fracture electron microscopy of reaggregated control cells showed interfaces containing an average of more than 600 aggregated intramembranous gap junction particles, while TPA-treated cells had no gap junctions. However, Lucifer yellow dye transfer between nondissociated cells via gap junctions was unaffected by 60 min of TPA treatment. Therefore, TPA dramatically inhibited gap junction assembly but did not alter channel gating nor enhance disassembly of preexisting gap junction structures. Short term TPA treatment (< 30 min) increased phosphorylation of the gap junction protein molecular weight of 43,000 (Cx43), but did not change the cellular level of Cx43. Cell surface biotinylation experiments suggested that TPA did not substantially reduce the plasma membrane concentration of Cx43. Therefore, the simple presence of Cx43 in the plasma membrane is not sufficient for gap junction assembly, and protein kinase C probably exerts an effect on assembly of gap junctions at the plasma membrane level. PMID:7806568

Production of Composite Scaffold Containing Silk Fibroin, Chitosan, and Gelatin for 3D Cell Culture and Bone Tissue Regeneration.

PubMed

Li, Jianqing; Wang, Qiuke; Gu, Yebo; Zhu, Yu; Chen, Liang; Chen, Yunfeng

2017-11-08

BACKGROUND Bone tissue engineering, a powerful tool to treat bone defects, is highly dependent on use of scaffolds. Both silk fibroin (SF) and chitosan (Cs) are biocompatible and actively studied for reconstruction of tissue engineering. Gelatin (Gel) is also widely applied in the biomedical field due to its low antigenicity and physicochemical stability. MATERIAL AND METHODS In this study, 4 different types of scaffolds were constructed - SF, SF/Cs, SF/Gel, and SF/Cs/Gel - and we compared their physical and chemical properties as well as biological characterization of these scaffolds to determine the most suitable scaffold for use in bone regeneration. First, these scaffolds were produced via chemical cross-linking method and freeze-drying technique. Next, the characterization of internal structure was studied using scanning electron microscopy and the porosity was evaluated by liquid displacement method. Then, we compared physicochemical properties such as water absorption rate and degradation property. Finally, MC3T3-E1 cells were inoculated on the scaffolds to study the biocompatibility and osteogenesis of the three-dimensional (3D) scaffolds in vitro. RESULTS The composite scaffold formed by all 3 components was the best for use in bone regeneration. CONCLUSIONS We conclude that the best scaffold among the 4 studied for MC3T3-E1 cells is our SF/Cs/Gel scaffold, suggesting a new choice for bone regeneration that can be used to treat bone defects or fractures in clinical practice.

The ginger component 6-shogaol prevents TNF-α-induced barrier loss via inhibition of PI3K/Akt and NF-κB signaling.

PubMed

Luettig, Julia; Rosenthal, Rita; Lee, In-Fah M; Krug, Susanne M; Schulzke, Jörg D

2016-12-01

Anti-inflammatory properties of the ginger-derived pungent component 6-shogaol (6-SG) have been studied intensively in recent years. Purpose of this study was to characterize the influence of 6-SG on inflammation-related intestinal barrier dysfunction, especially its paracellular component. The effect of 6-SG was studied in the human intestinal cell models HT-29/B6 and Caco-2 either under control conditions or challenged by the pro-inflammatory cytokine tumor necrosis factor α (TNF-α). Electrophysiological measurements, freeze-fracture electron microscopy, and protein analyses were performed. 6-SG partially prevented both, the TNF-α-induced decrease in transepithelial resistance and the rise in fluorescein permeability. By inhibiting phosphatidylinositol-3-kinase/Akt signaling 6-SG prevented the TNF-α-induced increase in protein expression of claudin-2, a channel-forming tight junction protein. In addition, the TNF-α-induced disassembly of the sealing tight junction protein claudin-1 was attenuated, the latter of which was due to TNF-α-triggered phosphorylation of nuclear factor kappa light chain enhancer of activated B cells (NF-κB). 6-SG has barrier-protective effects by affecting TNF-α-induced claudin-2 upregulation and claudin-1 disassembly via inhibition of phoshatidylinositol-3-kinase/Akt and nuclear factor kappa light chain enhancer of activated B-cell signaling. Therefore, 6-SG-containing food might be beneficial for barrier preservation during intestinal inflammation. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Physicochemical characterization of artificial nanoerythrosomes derived from erythrocyte ghost membranes.

PubMed

Deák, Róbert; Mihály, Judith; Szigyártó, Imola Cs; Wacha, András; Lelkes, Gábor; Bóta, Attila

2015-11-01

Colloidal stabile nanoerythrosomes with 200 nm average diameter were formed from hemoglobin-free erythrocyte ghost membrane via sonication and membrane extrusion. The incorporation of extra lipid (1,2-dipalmitoyl-sn-glycero-3-phosphocholine, DPPC), added to the sonicated ghosts, caused significant changes in the thermotropic character of the original membranes. As a result of the increased DPPC ratio the chain melting of the hydrated DPPC system and the characteristic small angle X-ray scattering (SAXS) of the lipid bilayers appeared. Significant morphological changes were followed by transmission electron microscopy combined with freeze fracture method (FF-TEM). After the ultrasonic treatment the large entities of erythrocyte ghosts transformed into nearly spherical nanoerythrosomes with diameters between 100 and 300 nm and at the same time a great number of 10-30 nm large membrane proteins or protein clusters were dispersed in the aqueous medium. The infrared spectroscopy (FT-IR) pointed out, that the sonication did not cause changes in the secondary structures of the membrane proteins under our preparation conditions. About fivefold of extra lipid--compared to the lipid content of the original membrane--caused homogeneous dispersion of nanoerythrosomes however the shape of the vesicles was not uniform. After the addition of about tenfold of DPPC, monoform and monodisperse nanoerythrosomes became typical. The outer surfaces of these roughly spherical objects were frequently polygonal, consisting of a net of pentagons and hexagons. Copyright © 2015 Elsevier B.V. All rights reserved.

Distinct cerebellar engrams in short-term and long-term motor learning.

PubMed

Wang, Wen; Nakadate, Kazuhiko; Masugi-Tokita, Miwako; Shutoh, Fumihiro; Aziz, Wajeeha; Tarusawa, Etsuko; Lorincz, Andrea; Molnár, Elek; Kesaf, Sebnem; Li, Yun-Qing; Fukazawa, Yugo; Nagao, Soichi; Shigemoto, Ryuichi

2014-01-07

Cerebellar motor learning is suggested to be caused by long-term plasticity of excitatory parallel fiber-Purkinje cell (PF-PC) synapses associated with changes in the number of synaptic AMPA-type glutamate receptors (AMPARs). However, whether the AMPARs decrease or increase in individual PF-PC synapses occurs in physiological motor learning and accounts for memory that lasts over days remains elusive. We combined quantitative SDS-digested freeze-fracture replica labeling for AMPAR and physical dissector electron microscopy with a simple model of cerebellar motor learning, adaptation of horizontal optokinetic response (HOKR) in mouse. After 1-h training of HOKR, short-term adaptation (STA) was accompanied with transient decrease in AMPARs by 28% in target PF-PC synapses. STA was well correlated with AMPAR decrease in individual animals and both STA and AMPAR decrease recovered to basal levels within 24 h. Surprisingly, long-term adaptation (LTA) after five consecutive daily trainings of 1-h HOKR did not alter the number of AMPARs in PF-PC synapses but caused gradual and persistent synapse elimination by 45%, with corresponding PC spine loss by the fifth training day. Furthermore, recovery of LTA after 2 wk was well correlated with increase of PF-PC synapses to the control level. Our findings indicate that the AMPARs decrease in PF-PC synapses and the elimination of these synapses are in vivo engrams in short- and long-term motor learning, respectively, showing a unique type of synaptic plasticity that may contribute to memory consolidation.

Effects of the Electron Beam Welding Process on the Microstructure, Tensile, Fatigue and Fracture Properties of Nickel Alloy Nimonic 80A

NASA Astrophysics Data System (ADS)

Zhang, H.; Huang, Chongxiang; Guan, Zhongwei; Li, Jiukai; Liu, Yongjie; Chen, Ronghua; Wang, Qingyuan

2018-01-01

The purpose of this study was to evaluate rotary bending high-cycle fatigue properties and crack growth of Nimonic 80A-based metal and electron beam-welded joints. All the tests were performed at room temperature. Fracture surfaces under high-cycle fatigue and fatigue crack growth were observed by scanning electron microscopy. Microstructure, hardness and tensile properties were also evaluated in order to understand the effects on the fatigue results obtained. It was found that the tensile properties, hardness and high-cycle fatigue properties of the welded joint are lower than the base metal. The fracture surface of the high-cycle fatigue shows that fatigue crack initiated from the surface under the high stress amplitude and from the subsurface under the low stress amplitude. The effect of the welding process on the statistical fatigue data was studied with a special focus on probabilistic life prediction and probabilistic lifetime limits. The fatigue crack growth rate versus stress intensity factor range data were obtained from the fatigue crack growth tests. From the results, it was evident that the fatigue crack growth rates of the welded are higher than the base metal. The mechanisms and fracture modes of fatigue crack growth of welded specimens were found to be related to the stress intensity factor range ΔK. In addition, the effective fatigue crack propagation thresholds and mismatch of welded joints were described and discussed.

Morphology Formation in PC/ABS Blends during Thermal Processing and the Effect of the Viscosity Ratio of Blend Partners

PubMed Central

Bärwinkel, Stefanie; Seidel, Andreas; Hobeika, Sven; Hufen, Ralf; Mörl, Michaela; Altstädt, Volker

2016-01-01

Morphology formation during compounding, as well as injection molding of blends containing 60 wt % polycarbonate (PC) and 40 wt % polybutadiene rubber-modified styrene-acrylonitrile copolymers (ABS), has been investigated by transmission electron microscopy (TEM). Profiles of the blend morphology have been recorded in injection-molded specimens and significant morphology gradients observed between their skin and core. A <10 µm thick surface layer with strongly dispersed and elongated nano-scale (streak-like) styrene acrylonitrile (SAN) phases and well-dispersed, isolated SAN-grafted polybutadiene rubber particles is followed by a 50–150 µm thick skin layer in which polymer morphology is characterized by lamellar SAN/ABS phases. Thickness of these lamellae increases with the distance from the specimen’s surface. In the core of the specimens the SAN-grafted polybutadiene rubber particles are exclusively present within the SAN phases, which exhibit a much coarser and less oriented, dispersed morphology compared to the skin. The effects of the viscosity of the SAN in the PC/ABS blends on phase morphologies and correlations with fracture mechanics in tensile and impact tests were investigated, including scanning electron microscopy (SEM) assessment of the fracture surfaces. A model explaining the mechanisms of morphology formation during injection molding of PC/ABS blends is discussed. PMID:28773780

Mechanical performance and thermal stability of glass fiber reinforced silica aerogel composites based on co-precursor method by freeze drying

NASA Astrophysics Data System (ADS)

Zhou, Ting; Cheng, Xudong; Pan, Yuelei; Li, Congcong; Gong, Lunlun; Zhang, Heping

2018-04-01

In order to maintain the integrity, glass fiber (GF) reinforced silica aerogel composites were synthesized using methltrimethoxysilane (MTMS) and water glass co-precursor by freeze drying method. The composites were characterized by scanning electron microscopy, Brunauer-Emmett-Teller analysis, uniaxial compressive test, three-point bending test, thermal conductivity analysis, contact angle test, TG-DSC analysis. It was found that the molar ratio of MTMS/water glass could significantly affect the properties of composites. The bulk density and thermal conductivity first decreased and then increased with the increasing molar ratio. The composites showed remarkable mechanical strength and flexibility compared with pure silica aerogel. Moreover, when the molar ratio is 1.8, the composites showed high specific surface area (870.9 m2/g), high contact angle (150°), great thermal stability (560 °C) and low thermal conductivity (0.0248 W/m·K). These outstanding properties indicate that GF/aerogels have broad prospects in the field of thermal insulation.

Supermacroporous chemically cross-linked poly(aspartic acid) hydrogels.

PubMed

Gyarmati, Benjámin; Mészár, E Zsuzsanna; Kiss, Lóránd; Deli, Mária A; László, Krisztina; Szilágyi, András

2015-08-01

Chemically cross-linked poly(aspartic acid) (PASP) gels were prepared by a solid-liquid phase separation technique, cryogelation, to achieve a supermacroporous interconnected pore structure. The precursor polymer of PASP, polysuccinimide (PSI) was cross-linked below the freezing point of the solvent and the forming crystals acted as templates for the pores. Dimethyl sulfoxide was chosen as solvent instead of the more commonly used water. Thus larger temperatures could be utilized for the preparation and the drawback of increase in specific volume of water upon freezing could be eliminated. The morphology of the hydrogels was characterized by scanning electron microscopy and interconnectivity of the pores was proven by the small flow resistance of the gels. Compression tests also confirmed the interconnected porous structure and the complete re-swelling and shape recovery of the supermacroporous PASP hydrogels. The prepared hydrogels are of interest for several biomedical applications as scaffolding materials because of their cytocompatibility, controllable morphology and pH-responsive character. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Freeze/thaw stress induces organelle remodeling and membrane recycling in cryopreserved human mature oocytes.

PubMed

Nottola, Stefania Annarita; Albani, Elena; Coticchio, Giovanni; Palmerini, Maria Grazia; Lorenzo, Caterina; Scaravelli, Giulia; Borini, Andrea; Levi-Setti, Paolo Emanuele; Macchiarelli, Guido

2016-12-01

Our aim was to evaluate the ultrastructure of human metaphase II oocytes subjected to slow freezing and fixed after thawing at different intervals during post-thaw rehydration. Samples were studied by light and transmission electron microscopy. We found that vacuolization was present in all cryopreserved oocytes, reaching a maximum in the intermediate stage of rehydration. Mitochondria-smooth endoplasmic reticulum (M-SER) aggregates decreased following thawing, particularly in the first and intermediate stages of rehydration, whereas mitochondria-vesicle (MV) complexes augmented in the same stages. At the end of rehydration, vacuoles and MV complexes both diminished and M-SER aggregates increased again. Cortical granules (CGs) were scarce in all cryopreserved oocytes, gradually diminishing as rehydration progressed. This study also shows that such a membrane remodeling is mainly represented by a dynamic process of transition between M-SER aggregates and MV complexes, both able of transforming into each other. Vacuoles and CG membranes may take part in the membrane recycling mechanism.

Fractographic Analysis of a Split Tooth Presenting Radiographically as a Horizontal Root Fracture in an Unrestored Mandibular Second Molar.

PubMed

Krishnan, Unni; Moule, Alex; Michael, Shaji; Swain, Michael

2018-02-01

Spontaneously catastrophic fracture of intact unrestored molar teeth is not common. Nevertheless, cracks do occur that progress apically, resulting in the complete splitting of the tooth and root. This report describes a catastrophic fracture that occurred in an unrestored mandibular second molar resulting in a previously unreported combination of a longitudinal and horizontal root fracture, appearing radiographically as a single horizontal root fracture. Tooth fragments were examined clinically, stereoscopically, and by scanning electron microscopy. Fractographic analysis was used to investigate the dynamics involved in fracture initiation, structural resistances encountered during progression of the fracture, and reasons for direction changes culminating in the unusual radiographic appearance. The uniqueness of this report is that it describes fractographic evidence of factors contributing to the initiation and progression of an in vivo crack. It shows fracture markings that are evidence of the energy dissipation mechanisms. The topographic location of these markings confirmed that cracks occur in vivo in stages with different rates of progression. This analysis helps to explain why split teeth are uncommon and highlights some of the multitude of factors that have to coincide for a tooth to catastrophically fracture. The report describes the mechanism of fracture and should stimulate clinicians and researchers to investigate cracking of teeth by undertaking fractographic analysis of extracted cracked teeth. Copyright © 2017 American Association of Endodontists. All rights reserved.

Contribution to the study of the vasculature of submandibular and sublingual glands and lymph nodes of rats by corrosion cast technique combined with scanning electron microscopy.

PubMed

Rossi-Schneider, Tíssiana Rachel; Verli, Flaviana Dornela; Yurgel, Liliane Soares; De Souza, Maria Antonieta Lopes; Cherubini, Karen

2008-10-01

The study of anatomical structures in their normal state allows the identification of pathological changes that can occur in them. Angiogenesis and the vasculature have been widely studied, mainly because of their association with the development of neoplasms. One of the methods applied for such purposes is the corrosion cast technique, which provides a copy of the vessels with normal as well as pathological structures. The replica of the vasculature provided by this technique allows the three-dimensional analysis of vessels by means of scanning electron microscopy. The aim of the present study was to demonstrate, by means of corrosion casts, the angioarchitecture of the submandibular and sublingual glands and lymph nodes. Scanning electron microscopy showed that the three structures have distinct vascular patterns. The corrosion cast technique can be employed in the study of the angioarchitecture of the submandibular and sublingual glands and lymph nodes, but requires specific precautions. The removal of the structures en bloc and the handling of the replicas with the aid of a stereoscopic magnifier reduce the risk of fractures. (c) 2008 Wiley-Liss, Inc.

Laser engravings as reason for mechanical failure of titanium-alloyed total hip stems.

PubMed

Kluess, Daniel; Steinhauser, Erwin; Joseph, Micheal; Koch, Ursula; Ellenrieder, Martin; Mittelmeier, Wolfram; Bader, Rainer

2015-07-01

Two revisions of broken β-titanium total hip stems had to be performed in our hospital after 2 and 4 years in situ. Since both fractures were located at the level of a laser engraving, a failure analysis was conducted. Both retrieved hip stems were disinfected and collected in our retrieval database after patient's signed agreement. Each fragment was macroscopically photographed. Fracture surfaces were analyzed using scanning electron microscopy (SEM). Quantification of element content was conducted using energy dispersive X-ray (EDX) analysis. Both stems show fatigue fracture, as displayed by the lines of rest on the fracture surface. The origin of fracture was identified directly at the laser engraving of the company logo at both stems by means of SEM. The EDX analysis showed an oxygen level beneath the laser engraving about twice as high as in the substrate, causing material embrittlement. Laser engravings need to be reduced to a minimum of necessary information, and should be placed at locations with minimum mechanical load. Biomechanical analyses are recommended to identify less loaded areas in implant components to avoid such implant failures.

Structure and Properties of the Aluminide Coatings on the Inconel 625 Superalloy

NASA Astrophysics Data System (ADS)

Adamiak, Stanisław; Bochnowski, Wojciech; Dziedzic, Andrzej; Filip, Ryszard; Szeregij, Eugeniusz

2016-01-01

The research samples used in this study were based on the Inconel 625 alloy; the examined samples were coated with aluminide films deposited in a low-activity chemical vapor deposition (CVD) process. The samples' microstructure was investigated with optical and electron microscopy and energy dispersive X-ray spectroscopy analysis. Hardness measurements were performed using Vickers and Berkovich test methods. The adhesion of the aluminide coating was determined by fractography. It was shown that the fracture mechanism was different for the respective zones of the aluminide coating and the substrate material. The outer zone of the aluminide coating is characterized by an intercrystalline fracture, with a small contribution of transcrystalline fracture within individual grains (large crystallites in the bottom of the zone, composed of smaller crystallites, also show an intercrystalline fracture). The substrate material exhibited a ductile intercrystalline fracture. Based on this investigation, an increase of the microhardness of the material occurring at loads below 0.2 N was observed. When determining microhardness of aluminide coating it is necessary to take into account the optimal choice of the indentation tip.

Enhanced properties of nanostructured TiO2-graphene composites by rapid sintering

NASA Astrophysics Data System (ADS)

Shon, In-Jin; Yoon, Jin-Kook; Hong, Kyung-Tae

2018-01-01

Despite of many attractive properties of TiO2, the drawback of TiO2 ceramic is low fracture toughness for widely industrial application. The method to improve the fracture toughness and hardness has been reported by addition of reinforcing phase to fabricate a nanostructured composite. In this regard, graphene has been evaluated as an ideal second phase in ceramics. Nearly full density of nanostructured TiO2-graphene composite was achieved within one min using pulsed current activated sintering. The effect of graphene on microstructure, fracture toughness and hardness of TiO2-graphene composite was evaluated using Vickers hardness tester and field emission scanning electron microscopy. The grain size of TiO2 in the TiO2-x vol% (x = 0, 1, 3, and 5) graphene composite was greatly reduced with increase in addition of graphene. Both hardness and fracture toughness of TiO2-graphene composites simultaneously increased in the addition of graphene.

Effects of metal binder on the microstructure and mechanical properties of Al2O3-based micro-nanocomposite ceramic tool material

NASA Astrophysics Data System (ADS)

Ni, Xiu-ying; Zhao, Jun; Sun, Jia-lin; Gong, Feng; Li, Zuo-li

2017-07-01

The Al2O3-(W,Ti)C composites with Ni and Mo additions varying from 0vol% to 12vol% were prepared via hot pressing sintering under 30 MPa. The microstructure was investigated via X-ray diffraction (XRD) and scanning electron microscopy (SEM) equipped with energy dispersive spectrometry (EDS). Mechanical properties such as flexural strength, fracture toughness, and Vickers hardness were also measured. Results show that the main phases A12O3 and (W,Ti)C were detected by XRD. Compound MoNi also existed in sintered nanocomposites. The fracture modes of the nanocomposites were both intergranular and transgranular fractures. The plastic deformation of metal particles and crack bridging were the main toughening mechanisms. The maximum flexural strength and fracture toughness were obtained for 9vol% and 12vol% additions of Ni and Mo, respectively. The hardness of the composites reduced gradually with increasing content of metals Ni and Mo.

Low cycle fatigue behavior of a ferritic reactor pressure vessel steel

NASA Astrophysics Data System (ADS)

Sarkar, Apu; Kumawat, Bhupendra K.; Chakravartty, J. K.

2015-07-01

The cyclic stress-strain response and the low cycle fatigue (LCF) behavior of 20MnMoNi55 pressure vessel steel were studied. Tensile strength and LCF properties were examined at room temperature (RT) using specimens cut from rolling direction of a rolled block. The fully reversed strain-controlled LCF tests were conducted at a constant total strain rate with different axial strain amplitude levels. The cyclic strain-stress relationships and the strain-life relationships were obtained through the test results, and related LCF parameters of the steel were calculated. The studied steel exhibits cyclic softening behavior. Furthermore, analysis of stabilized hysteresis loops showed that the steel exhibits non-Masing behavior. Complementary scanning electron microscopy examinations were also carried out on fracture surfaces to reveal dominant damage mechanisms during crack initiation, propagation and fracture. Multiple crack initiation sites were observed on the fracture surface. The investigated LCF behavior can provide reference for pressure vessel life assessment and fracture mechanisms analysis.

High damage tolerance of electrochemically lithiated silicon

DOE PAGES

Wang, Xueju; Fan, Feifei; Wang, Jiangwei; ...

2015-09-24

Mechanical degradation and resultant capacity fade in high-capacity electrode materials critically hinder their use in high-performance rechargeable batteries. Despite tremendous efforts devoted to the study of the electro–chemo–mechanical behaviours of high-capacity electrode materials, their fracture properties and mechanisms remain largely unknown. In this paper, we report a nanomechanical study on the damage tolerance of electrochemically lithiated silicon. Our in situ transmission electron microscopy experiments reveal a striking contrast of brittle fracture in pristine silicon versus ductile tensile deformation in fully lithiated silicon. Quantitative fracture toughness measurements by nanoindentation show a rapid brittle-to-ductile transition of fracture as the lithium-to-silicon molar ratiomore » is increased to above 1.5. Molecular dynamics simulations elucidate the mechanistic underpinnings of the brittle-to-ductile transition governed by atomic bonding and lithiation-induced toughening. Finally, our results reveal the high damage tolerance in amorphous lithium-rich silicon alloys and have important implications for the development of durable rechargeable batteries.« less

Size-segregated urban aerosol characterization by electron microscopy and dynamic light scattering and influence of sample preparation

NASA Astrophysics Data System (ADS)

Marvanová, Soňa; Kulich, Pavel; Skoupý, Radim; Hubatka, František; Ciganek, Miroslav; Bendl, Jan; Hovorka, Jan; Machala, Miroslav

2018-04-01

Size-segregated particulate matter (PM) is frequently used in chemical and toxicological studies. Nevertheless, toxicological in vitro studies working with the whole particles often lack a proper evaluation of PM real size distribution and characterization of agglomeration under the experimental conditions. In this study, changes in particle size distributions during the PM sample manipulation and also semiquantitative elemental composition of single particles were evaluated. Coarse (1-10 μm), upper accumulation (0.5-1 μm), lower accumulation (0.17-0.5 μm), and ultrafine (<0.17 μm) PM fractions were collected by high volume cascade impactor in Prague city center. Particles were examined using electron microscopy and their elemental composition was determined by energy dispersive X-ray spectroscopy. Larger or smaller particles, not corresponding to the impaction cut points, were found in all fractions, as they occur in agglomerates and are impacted according to their aerodynamic diameter. Elemental composition of particles in size-segregated fractions varied significantly. Ns-soot occurred in all size fractions. Metallic nanospheres were found in accumulation fractions, but not in ultrafine fraction where ns-soot, carbonaceous particles, and inorganic salts were identified. Dynamic light scattering was used to measure particle size distribution in water and in cell culture media. PM suspension of lower accumulation fraction in water agglomerated after freezing/thawing the sample, and the agglomerates were disrupted by subsequent sonication. Ultrafine fraction did not agglomerate after freezing/thawing the sample. Both lower accumulation and ultrafine fractions were stable in cell culture media with fetal bovine serum, while high agglomeration occurred in media without fetal bovine serum as measured during 24 h.

Image-Guided Cryoablation of the Spine in a Swine Model: Clinical, Radiological, and Pathological Findings with Light and Electron Microscopy.

PubMed

de Freitas, Ricardo Miguel Costa; Andrade, Celi Santos; Caldas, José Guilherme Mendes Pereira; Tsunemi, Miriam Harumi; Ferreira, Lorraine Braga; Arana-Chavez, Victor Elias; Cury, Patrícia Maluf

2015-10-01

This study was designed to present the feasibility of an in vivo image-guided percutaneous cryoablation of the porcine vertebral body. The institutional animal care committee approved this study. Cone-beam computed tomography (CBCT)-guided vertebral cryoablations (n = 22) were performed in eight pigs with short, 2-min, single or double-freezing protocols. Protective measures to nerves included dioxide carbon (CO2) epidural injections and spinal canal temperature monitoring. Clinical, radiological, and pathological data with light (n = 20) or transmission electron (n = 2) microscopic analyses were evaluated after 6 days of clinical follow-up and euthanasia. CBCT/fluoroscopic-guided transpedicular vertebral body cryoprobe positioning and CO2 epidural injection were successful in all procedures. No major complications were observed in seven animals (87.5 %, n = 8). A minor complication was observed in one pig (12.5 %, n = 1). Logistic regression model analysis showed the cryoprobe-spinal canal (Cp-Sc) distance as the most efficient parameter to categorize spinal canal temperatures lower than 19 °C (p < 0.004), with a significant Pearson's correlation test (p < 0.041) between the Cp-Sc distance and the lowest spinal canal temperatures. Ablation zones encompassed pedicles and the posterior wall of the vertebral bodies with an inflammatory rim, although no inflammatory infiltrate was depicted in the surrounding neural structures at light microscopy. Ultrastructural analyses evidenced myelin sheath disruption in some large nerve fibers, although neurological deficits were not observed. CBCT-guided vertebral cryoablation of the porcine spine is feasible under a combination of a short freezing protocol and protective measures to the surrounding nerves. Ultrastructural analyses may be helpful assess the early modifications of the nerve fibers.

Scanning electron microscopy of echinoid podia.

PubMed

Florey, E; Cahill, M A

1982-01-01

Tube feet of the sea urchin Strongylocentrotus franciscanus were studied with the scanning electron microscope (SEM). By use of fractured preparations it was possible to obtain views of all components of the layered tube-foot wall. The outer epithelium was found to bear tufts of cilia possibly belonging to sensory cells. The nerve plexus was clearly revealed as being composed of bundles of varicose axons. The basal lamina, which covers the outer and inner surfaces of the connective tissue layer, was found to be a mechanically resistant and elastic membrane. The connective tissue appears as dense bundles of (collagen) fibers. The luminal epithelium (coelothelium) is a single layer of flagellated collar cells. There is no indication that the muscle fibers, which insert on the inner basal lamina of the connective tissue layer are innervated by axons from the basi-epithelial nerve plexus. The results agree with previous conclusions concerning tube-foot structure based on transmission electron microscopy, and provide additional information, particularly with regard to the outer and inner epithelia.

Depositing nanoparticles on a silicon substrate using a freeze drying technique.

PubMed

Sigehuzi, Tomoo

2017-08-28

For the microscopic observation of nanoparticles, an adequate sample preparation is an essential part of this task. Much research has been performed for usable preparation methods that will yield aggregate-free samples. A freeze drying technique, which only requires a -80  ° C freezer and a freeze dryer, is shown to provide an on-substrate dispersion of mostly isolated nanoparticles. The particle density could be made sufficiently high for efficient observations using atomic force microscopy. Since this sandwich method is purely physical, it could be applied to deposit various nanoparticles independent of their surface chemical properties. Suspension film thickness, or the dimensionality of the suspension film, was shown to be crucial for the isolation of the particles. Silica nanoparticles were dispersed on a silicon substrate using this method and the sample properties were examined using atomic force microscopy.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Souza, Douglas F., E-mail: souzadf@outlook.com; Nunes, Eduardo H.M., E-mail: eduardohmn@gmail.com; Pimenta, Daiana S.

In this work we fabricated alumina samples by the freeze-casting technique using tert-butanol as the solvent. The prepared materials were examined by scanning electron microscopy and X-ray microtomography. Next, they were coated with sol–gel silica films by dip-coating. Permeability tests were carried out in order to assess the permeation behavior of the materials processed in this study. We observed that the sintering time and alumina loading showed a remarkable effect on both the structural properties and flexural strength of the freeze-cast samples. Nitrogen adsorption tests revealed that the silica prepared in this study exhibited a microporous structure. It was observedmore » that the presence of silica coatings on the alumina surface decreased the CO{sub 2} permeance by about one order of magnitude. Because of the similar kinetic diameters of nitrogen and carbon dioxide, the CO{sub 2}/N{sub 2} system showed a separation efficiency that was lower than that observed for the He/CO{sub 2} and He/N{sub 2} systems. We noticed that increasing the feed pressure improved the separation capacity of the obtained materials. - Highlights: • Porous alumina samples obtained by the freeze-casting technique • Microporous silica coating prepared by a simple sol–gel dip-coating methodology • Samples examined by SEM, μ-CT, and nitrogen sorption tests • Mechanical tests were carried out in the freeze-cast samples. • The presence of silica coatings on the alumina surface decreased the CO{sub 2} permeance.« less

Sol-gel NiFe2O4 nanoparticles: Effect of the silica coating

NASA Astrophysics Data System (ADS)

Larumbe, S.; Pérez-Landazábal, J. I.; Pastor, J. M.; Gómez-Polo, C.

2012-05-01

NiFe2O4 and NiFe2O4-SiO2 nanoparticles were synthesized by a sol-gel method using citric acid as fuel, giving rise its combustion to the crystallization of the spinel phase. Different synthesis conditions were analyzed with the aim of obtaining stoichiometric NiFe2O4 nanoparticles. The spinel structure in the calcined nanoparticles (400 °C, 2 h) was evaluated by x-ray diffraction. Their nanometer size (mean diameters around 10-15 nm) was confirmed through electron microscopy (field emission scanning electron microscopy and transmission electron microscopy). Rietveld refinement indicates the existence of a small percentage of NiO and Fe3O4 phases and a certain degree of structural disorder. The main effect of the silica coating is to enhance the disorder effects and prevent the crystalline growth after post-annealing treatments. Due to the small particle size, the nanoparticles display characteristic superparamagnetic behaviour and surface effects associated to a spin-glass like state: i.e., reduction in the saturation magnetization values and splitting of the zero field cooled (ZFC)-field cooled (FC) high field magnetization curves. The fitting of the field dependence of the ZFC-FC irreversibility temperatures to the Almeida—Thouless equation confirms the spin-glass nature of the detected magnetic phenomena. Exchange bias effects (shifts in the FC hysteresis loops) detected below the estimated freezing temperature support the spin-glass nature of the spin disorder effects.

Mineral Precipitation in Fractures: Multiscale Imaging and Geochemical Modeling

NASA Astrophysics Data System (ADS)

Hajirezaie, S.; Peters, C. A.; Swift, A.; Sheets, J. M.; Cole, D. R.; Crandall, D.; Cheshire, M.; Stack, A. G.; Anovitz, L. M.

2017-12-01

For subsurface energy technologies such as geologic carbon sequestration, fractures are potential pathways for fluid migration from target formations. Highly permeable fractures may become sealed by mineral precipitation. In this study, we examined shale specimens with existing cemented fractures as natural analogues, using an array of imaging methods to characterize mineralogy and porosity at several spatial scales. In addition, we used reactive transport modeling to investigate geochemical conditions that can lead to extensive mineral precipitation and to simulate the impacts on fracture hydraulic properties. The naturally-cemented fractured rock specimens were from the Upper Wolfcamp formation in Texas, at 10,000 ft depth. The specimens were scanned using x-ray computed tomography (xCT) at resolution of 13 microns. The xCT images revealed an original fracture aperture of 1.9 mm filled with several distinct mineral phases and vuggy void regions, and the mineral phase volumes and surface areas were quantified and mapped in 3D. Specimens were thin-sectioned and examined at micron- and submicron-scales using petrographic microscopy (PM), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDX), and small angle X-ray scattering (SAXS). Collectively these methods revealed crystals of dolomite as large as 900 microns in length overlain with a heterogeneous mixture of carbonate minerals including calcite, dolomite, and Fe-rich dolomite, interspersed at spatial scales as small as 5 microns. In addition, secondary precipitation of SiO2 was found to fill some of the void space. This multiscale imaging was used to inform the reactive transport modeling employed to examine the conditions that can cause the observed mineral precipitation in fractures at a larger scale. Two brines containing solutions that when mixed would lead to precipitation of various carbonate minerals were simulated as injectants into a fracture domain. In particular, the competing effects of transport dynamics and reaction kinetics were investigated in the context of profiles of the precipitated minerals and permeability behavior of the fracture flow path. This study contributes rich knowledge toward mastering the subsurface for energy production and storage and for the management of energy waste streams.

Analysis of Fracture Mechanism for Al-Mg/SiCp Composite Materials

NASA Astrophysics Data System (ADS)

Maleque, M. A.; Adebisi, A. A.; Izzati, N.

2017-03-01

The present study aims to examine the fracture mechnism of silicon carbide particle (SiCp) reinforced aluminium matrix composite (AMC) material with 1 wt% addition of magnesium is fabricated using the stir casting process. The aluminium composite (Al-Mg/SiCp) is investigated for fatigue life and impact strength considering reinforcement weight fraction and influence of temperature on fracture toughness. The fabricated composite was tested using fatigue testing machine and charpy impact tester. Fractographic observations were evaluated with the scanning electron microscopy (SEM) on the fracture surface. It was found that increasing the SiCp weight fraction increased the fatigue life of the composite. Moreover, the 20 wt% SiCp Al-Mg composite attained the highest number of cycle and fatigue life compared to other variations. The mechanism responsible for the phenomena includes load transfer from the Al matrix alloy phase to the high strength and stiffness of the incorporated SiCp. The temperature variation influenced the impact strength of the composite and improved fracture toughness is achieved at 150 °C. It can be concluded from this study that reinforcement weight fraction and temperature affects the fracture behavior of the composites.

Dilatational band formation in bone

PubMed Central

Poundarik, Atharva A.; Diab, Tamim; Sroga, Grazyna E.; Ural, Ani; Boskey, Adele L.; Gundberg, Caren M.; Vashishth, Deepak

2012-01-01

Toughening in hierarchically structured materials like bone arises from the arrangement of constituent material elements and their interactions. Unlike microcracking, which entails micrometer-level separation, there is no known evidence of fracture at the level of bone’s nanostructure. Here, we show that the initiation of fracture occurs in bone at the nanometer scale by dilatational bands. Through fatigue and indentation tests and laser confocal, scanning electron, and atomic force microscopies on human and bovine bone specimens, we established that dilatational bands of the order of 100 nm form as ellipsoidal voids in between fused mineral aggregates and two adjacent proteins, osteocalcin (OC) and osteopontin (OPN). Laser microdissection and ELISA of bone microdamage support our claim that OC and OPN colocalize with dilatational bands. Fracture tests on bones from OC and/or OPN knockout mice (OC−/−, OPN−/−, OC-OPN−/−;−/−) confirm that these two proteins regulate dilatational band formation and bone matrix toughness. On the basis of these observations, we propose molecular deformation and fracture mechanics models, illustrating the role of OC and OPN in dilatational band formation, and predict that the nanometer scale of tissue organization, associated with dilatational bands, affects fracture at higher scales and determines fracture toughness of bone. PMID:23129653

What transmission electron microscopes can visualize now and in the future.

PubMed

Müller, Shirley A; Aebi, Ueli; Engel, Andreas

2008-09-01

Our review concentrates on the progress made in high-resolution transmission electron microscopy (TEM) in the past decade. This includes significant improvements in sample preparation by quick-freezing aimed at preserving the specimen in a close-to-native state in the high vacuum of the microscope. Following advances in cold stage and TEM vacuum technology systems, the observation of native, frozen hydrated specimens has become a widely used approach. It fostered the development of computer guided, fully automated low-dose data acquisition systems allowing matched pairs of images and diffraction patterns to be recorded for electron crystallography, and the collection of entire tilt-series for electron tomography. To achieve optimal information transfer to atomic resolution, field emission electron guns combined with acceleration voltages of 200-300 kV are now routinely used. The outcome of these advances is illustrated by the atomic structure of mammalian aquaporin-O and by the pore-forming bacterial cytotoxin ClyA resolved to 12 A. Further, the Yersinia injectisome needle, a bacterial pseudopilus and the binding of phalloidin to muscle actin filaments were chosen to document the advantage of the high contrast offered by dedicated scanning transmission electron microscopy (STEM) and/or the STEM's ability to measure the mass of protein complexes and directly link this to their shape. Continued progress emerging from leading research laboratories and microscope manufacturers will eventually enable us to determine the proteome of a single cell by electron tomography, and to more routinely solve the atomic structure of membrane proteins by electron crystallography.

Comparison of the fracture resistances of glass fiber mesh- and metal mesh-reinforced maxillary complete denture under dynamic fatigue loading.

PubMed

Im, So-Min; Huh, Yoon-Hyuk; Cho, Lee-Ra; Park, Chan-Jin

2017-02-01

The aim of this study was to investigate the effect of reinforcing materials on the fracture resistances of glass fiber mesh- and Cr-Co metal mesh-reinforced maxillary complete dentures under fatigue loading. Glass fiber mesh- and Cr-Co mesh-reinforced maxillary complete dentures were fabricated using silicone molds and acrylic resin. A control group was prepared with no reinforcement (n = 15 per group). After fatigue loading was applied using a chewing simulator, fracture resistance was measured by a universal testing machine. The fracture patterns were analyzed and the fractured surfaces were observed by scanning electron microscopy. After cyclic loading, none of the dentures showed cracks or fractures. During fracture resistance testing, all unreinforced dentures experienced complete fracture. The mesh-reinforced dentures primarily showed posterior framework fracture. Deformation of the all-metal framework caused the metal mesh-reinforced denture to exhibit the highest fracture resistance, followed by the glass fiber mesh-reinforced denture ( P <.05) and the control group ( P <.05). The glass fiber mesh-reinforced denture primarily maintained its original shape with unbroken fibers. River line pattern of the control group, dimples and interdendritic fractures of the metal mesh group, and radial fracture lines of the glass fiber group were observed on the fractured surfaces. The glass fiber mesh-reinforced denture exhibits a fracture resistance higher than that of the unreinforced denture, but lower than that of the metal mesh-reinforced denture because of the deformation of the metal mesh. The glass fiber mesh-reinforced denture maintains its shape even after fracture, indicating the possibility of easier repair.

Nano-scale zero valent iron transport in a variable aperture dolomite fracture and a glass fracture

NASA Astrophysics Data System (ADS)

Mondal, P.; Sleep, B. E.; Cui, Z.; Zhou, Z.

2014-12-01

Experiments and numerical simulations are being performed to understand the transport behavior of carboxymethyl cellulose polymer stabilized nano-scale zero valent iron (nZVI) in a variable aperture dolomite rock fracture and a variable aperture glass replica of a fractured slate. The rock fracture was prepared by artificially inducing a fracture in a dolomite block along a stylolite, and the glass fracture was prepared by creating molds with melted glass on two opposing sides of a fractured slate rock block. Both of the fractures were 0.28 m in length and 0.21 m in width. Equivalent hydraulic apertures are about 110 microns for the rock fracture and 250 microns for the glass replica fracture. Sodium bromide and lissamine green B (LGB) serve as conservative tracers in the rock fracture and glass replica fracture, respectively. A dark box set-up with a light source and digital camera is being used to visualize the LGB and CMC-nZVI movement in the glass fracture. Experiments are being performed to determine the effects of water specific discharge and CMC concentration on nZVI transport in the fractures. Transmission electron microscopy, dynamic light scattering, and UV-visual spectrophotometry were performed to determine the stability and characteristics of the CMC-nZVI mixture. The transport of bromide, LGB, CMC, and CMC-nZVI in both fractures is being evaluated through analysis of the effluent concentrations. Time-lapse images are also being captured for the glass fracture. Bromide, LGB, and CMC recoveries have exceeded 95% in both fractures. Significant channeling has been observed in the fractures for CMC transport due to viscous effects.

Two-dimensional freezing criteria for crystallizing colloidal monolayers

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang Ziren; Han Yilong; Alsayed, Ahmed M.

Video microscopy was employed to explore crystallization of colloidal monolayers composed of diameter-tunable microgel spheres. Two-dimensional (2D) colloidal liquids were frozen homogenously into polycrystalline solids, and four 2D criteria for freezing were experimentally tested in thermal systems for the first time: the Hansen-Verlet freezing rule, the Loewen-Palberg-Simon dynamical freezing criterion, and two other rules based, respectively, on the split shoulder of the radial distribution function and on the distribution of the shape factor of Voronoi polygons. Importantly, these freezing criteria, usually applied in the context of single crystals, were demonstrated to apply to the formation of polycrystalline solids. At themore » freezing point, we also observed a peak in the fluctuations of the orientational order parameter and a percolation transition associated with caged particles. Speculation about these percolated clusters of caged particles casts light on solidification mechanisms and dynamic heterogeneity in freezing.« less

The contribution of apoptosis and necrosis in freezing injury of sea urchin embryonic cells.

PubMed

Boroda, Andrey V; Kipryushina, Yulia O; Yakovlev, Konstantin V; Odintsova, Nelly A

2016-08-01

Sea urchins have recently been reported to be a promising tool for investigations of oxidative stress, UV light perturbations and senescence. However, few available data describe the pathway of cell death that occurs in sea urchin embryonic cells after cryopreservation. Our study is focused on the morphological and functional alterations that occur in cells of these animals during the induction of different cell death pathways in response to cold injury. To estimate the effect of cryopreservation on sea urchin cell cultures and identify the involved cell death pathways, we analyzed cell viability (via trypan blue exclusion test, MTT assay and DAPI staining), caspase activity (via flow cytometry and spectrophotometry), the level of apoptosis (via annexin V-FITC staining), and cell ultrastructure alterations (via transmission electron microscopy). Using general caspase detection, we found that the level of caspase activity was low in unfrozen control cells, whereas the number of apoptotic cells with activated caspases rose after freezing-thawing depending on cryoprotectants used, also as the number of dead cells and cells in a late apoptosis. The data using annexin V-binding assay revealed a very high apoptosis level in all tested samples, even in unfrozen cells (about 66%). Thus, annexin V assay appears to be unsuitable for sea urchin embryonic cells. Typical necrotic cells with damaged mitochondria were not detected after freezing in sea urchin cell cultures. Our results assume that physical cell disruption but not freezing-induced apoptosis or necrosis is the predominant reason of cell death in sea urchin cultures after freezing-thawing with any cryoprotectant combination. Copyright © 2016 Elsevier Inc. All rights reserved.

Dynamic Torsional and Cyclic Fracture Behavior of ProFile Rotary Instruments at Continuous or Reciprocating Rotation as Visualized with High-speed Digital Video Imaging.

PubMed

Tokita, Daisuke; Ebihara, Arata; Miyara, Kana; Okiji, Takashi

2017-08-01

This study examined the dynamic fracture behavior of nickel-titanium rotary instruments in torsional or cyclic loading at continuous or reciprocating rotation by means of high-speed digital video imaging. The ProFile instruments (size 30, 0.06 taper; Dentsply Maillefer, Ballaigues, Switzerland) were categorized into 4 groups (n = 7 in each group) as follows: torsional/continuous (TC), torsional/reciprocating (TR), cyclic/continuous (CC), and cyclic/reciprocating (CR). Torsional loading was performed by rotating the instruments by holding the tip with a vise. For cyclic loading, a custom-made device with a 38° curvature was used. Dynamic fracture behavior was observed with a high-speed camera. The time to fracture was recorded, and the fractured surface was examined with scanning electron microscopy. The TC group initially exhibited necking of the file followed by the development of an initial crack line. The TR group demonstrated opening and closing of a crack according to its rotation in the cutting and noncutting directions, respectively. The CC group separated without any detectable signs of deformation. In the CR group, initial crack formation was recognized in 5 of 7 samples. The reciprocating rotation exhibited a longer time to fracture in both torsional and cyclic fatigue testing (P < .05). The scanning electron microscopic images showed a severely deformed surface in the TR group. The dynamic fracture behavior of NiTi rotary instruments, as visualized with high-speed digital video imaging, varied between the different modes of rotation and different fatigue testing. Reciprocating rotation induced a slower crack propagation and conferred higher fatigue resistance than continuous rotation in both torsional and cyclic loads. Copyright © 2017 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Fractographic analysis of 2.0-mm plates with a screw locking system in simulated fractures of the mandibular body.

PubMed

de Medeiros, Raquel Correia; Lauria de Moura, Andrezza; Rodrigues, Danillo Costa; Menezes Mendes, Marcelo Breno; Sawazaki, Renato; Fernandes Moreira, Roger William

2014-06-01

The purpose of the present study was to analyze the fractured plates from 2 brands of 2.0-mm locking fixation systems submitted to axial linear load testing. Four aluminum hemimandibles with linear sectioning to simulate a mandibular body fracture were used as a substrate and fixed with 2 fixation techniques from 2 national brands: Tóride and Traumec. The techniques were as follows: one 4-hole plate, with four 6-mm screws in the tension zone, and one 4-hole plate, with four 10-mm screws in the compression zone; and one 4-hole plate, with four 6-mm holes in the neutral zone. The hemimandibles were submitted to vertical linear load tests using an Instron 4411 mechanical test machine. The system was submitted to the test until complete failure had occurred. Next, a topographic analysis of the surface of the plates was performed using a stereomicroscope and an electronic scanning microscope. The samples were evaluated using different magnifications, and images were obtained. The surface of the fracture analyzed in scanning electron microscopy demonstrated a ductile-type fracture, usually found in the traction test bodies of ductile materials, such as titanium. No evidence of failure was observed in any fracture surface from a change in the structure or composition of the material. The plates were fractured by a ductile rupture mechanism, as expected, suggesting that the manufacturing of the national brand name plates used in the present study has been under adequate quality control, with no structural changes produced by the manufacturing process that could compromise their function. Copyright © 2014 American Association of Oral and Maxillofacial Surgeons. Published by Elsevier Inc. All rights reserved.

Mechanical properties and fracture toughness of rail steels and thermite welds at low temperature

NASA Astrophysics Data System (ADS)

Wang, Yuan-qing; Zhou, Hui; Shi, Yong-jiu; Feng, Bao-rui

2012-05-01

Brittle fracture occurs frequently in rails and thermite welded joints, which intimidates the security and reliability of railway service. Railways in cold regions, such as Qinghai-Tibet Railway, make the problem of brittle fracture in rails even worse. A series of tests such as uniaxial tensile tests, Charpy impact tests, and three-point bending tests were carried out at low temperature to investigate the mechanical properties and fracture toughness of U71Mn and U75V rail steels and their thermite welds. Fracture micromechanisms were analyzed by scanning electron microscopy (SEM) on the fracture surfaces of the tested specimens. The ductility indices (percentage elongation after fracture and percentage reduction of area) and the toughness indices (Charpy impact energy A k and plane-strain fracture toughness K IC) of the two kinds of rail steels and the corresponding thermite welds all decrease as the temperature decreases. The thermite welds are more critical to fracture than the rail steel base metals, as indicated by a higher yield-to-ultimate ratio and a much lower Charpy impact energy. U71Mn rail steel is relatively higher in toughness than U75V, as demonstrated by larger A k and K IC values. Therefore, U71Mn rail steel and the corresponding thermite weld are recommended in railway construction and maintenance in cold regions.

Wear effects on microscopic morphology and hyaluronan uptake in siloxane-hydrogel contact lenses.

PubMed

Tavazzi, Silvia; Tonveronachi, Martina; Fagnola, Matteo; Cozza, Federica; Ferraro, Lorenzo; Borghesi, Alessandro; Ascagni, Miriam; Farris, Stefano

2015-07-01

The purpose of this study was a comparison between new and worn siloxane-hydrogel contact lenses in terms of microscopic structure, surface morphology, and loading of hyaluronan. The analyses were performed by scanning electron microscopy, with the support of the freeze-drying technique, and by fluorescence confocal microscopy. Along the depth profile of new lenses, a thin porous top layer was observed, which corresponds to the region of hyaluronan penetration inside well-defined channels. The time evolution was followed from one day to two weeks of daily wear, when a completely different scenario was found. Clear experimental evidence of a buggy surface was observed with several crests and regions of swelling, which could be filled by the hyaluronan solution. The modifications are attributed to the progressive relaxation of the structure of the polymeric network. © 2014 Wiley Periodicals, Inc.

Preparation of Degradable Biological Carrier With LCC and its Application in Culture of Hepatocytes

NASA Astrophysics Data System (ADS)

Zhao, H. K.; Chen, X. K.; Wu, H. F.; Li, J. L.; Xie, Y. M.

2018-05-01

The purpose of this article is to extract lignin-carbohydrate complexes (LCC) with poplar as raw material, which was used to prepare bio-carrier by freeze-drying method. The chemical properties and morphological of LCC porous biological carriers were analyzed by GPC, FT-IR, scanning electron microscopy (SEM) and optical microscopy. The FT-IR spectrum results indicated that LCC which are composed of lignin and polysaccharide, with a typical LCC structure. Galactose have a specific ability to recognize liver cells owing to the presence of receptors on hepatocytes. Cell counting results showed that the cells increases fastest while the proliferation rate of the liver cell in LCC is obviously higher than that of control group. These results indicated that poplar LCC is very biocompatible, in which it might be a great potential biological carrier material for human hepatocyte culture.

Anther development of maize (Zea mays) and longstamen rice (Oryza longistaminata) revealed by cryo-SEM, with foci on locular dehydration and pollen arrangement.

PubMed

Tsou, Chih-Hua; Cheng, Ping-Chin; Tseng, Chiung-Maan; Yen, Hsiao-Jung; Fu, Yu-Lan; You, Tien-Rong; Walden, David B

2015-03-01

Key message: Pollen maturation in Poaceae. Another development has been extensively examined by various imaging tools, including transmission electron microscopy, scanning electron microscopy, and light microscopy, but none is capable of identifying liquid water. Cryo-scanning electron microscopy with high-pressure rapid freeze fixation is excellent in preserving structures at cellular level and differentiating gas- versus liquid-filled space, but rarely used in anther study. We applied this technique to examine anther development of Poaceae because of its economic importance and unusual peripheral arrangement of pollen. Maize and longstamen rice were focused on. Here, we report for the first time that anthers of Poaceae lose the locular free liquid during late-microspore to early pollen stages; the majority of pollen grains arranged in a tight peripheral whorl develops normally and reaches maturity in the gas-filled loculus. Occasionally, pollen grains are found situated in the locular cavity, but they remain immature or become shrunk at anthesis. At pollen stage, microchannels and cytoplasmic strands are densely distributed in the entire pollen exine and intine, respectively, suggesting that nutrients are transported into the pollen from the entire surface. We propose that in Poaceae, the specialized peripheral arrangement of pollen grains is crucial for pollen maturation in the gas-filled loculus, which enables pollen achieving large surface contact area with the tapetum and neighboring grains to maintain sufficient nutrient flow. This report also shows that the single aperture of pollen in Poaceae usually faces the tapetum, but other orientation is also common; pollen grains with different aperture orientations show no morphological differences.

Unconventional Imaging Methods to Capture Transient Structures during Actomyosin Interaction.

PubMed

Katayama, Eisaku; Kodera, Noriyuki

2018-05-08

Half a century has passed since the cross-bridge structure was recognized as the molecular machine that generates muscle tension. Despite various approaches by a number of scientists, information on the structural changes in the myosin heads, particularly its transient configurations, remains scant even now, in part because of their small size and rapid stochastic movements during the power stroke. Though progress in cryo-electron microscopy is eagerly awaited as the ultimate means to elucidate structural details, the introduction of some unconventional methods that provide high-contrast raw images of the target protein assemblies is quite useful, if available, to break the current impasse. Quick-freeze deep⁻etch⁻replica electron microscopy coupled with dedicated image analysis procedures, and high-speed atomic-force microscopy are two such candidates. We have applied the former to visualize actin-associated myosin heads under in vitro motility assay conditions, and found that they take novel configurations similar to the SH1⁻SH2-crosslinked myosin that we characterized recently. By incorporating biochemical and biophysical results, we have revised the cross-bridge mechanism to involve the new conformer as an important main player. The latter “microscopy” is unique and advantageous enabling continuous observation of various protein assemblies as they function. Direct observation of myosin-V’s movement along actin filaments revealed several unexpected behaviors such as foot-stomping of the leading head and unwinding of the coiled-coil tail. The potential contribution of these methods with intermediate spatial resolution is discussed.

Novel mesostructured inclusions in the epidermal lining of Artemia franciscana ovisacs show optical activity

PubMed Central

Hollergschwandtner, Elena; Schwaha, Thomas; Neumüller, Josef; Kaindl, Ulrich; Gruber, Daniela; Eckhard, Margret; Stöger-Pollach, Michael

2017-01-01

Background Biomineralization, e.g., in sea urchins or mollusks, includes the assembly of mesoscopic superstructures from inorganic crystalline components and biopolymers. The resulting mesocrystals inspire biophysicists and material scientists alike, because of their extraordinary physical properties. Current efforts to replicate mesocrystal synthesis in vitro require understanding the principles of their self-assembly in vivo. One question, not addressed so far, is whether intracellular crystals of proteins can assemble with biopolymers into functional mesocrystal-like structures. During our electron microscopy studies into Artemia franciscana (Crustacea: Branchiopoda), we found initial evidence of such proteinaceous mesostructures. Results EM preparations with high-pressure freezing and accelerated freeze substitution revealed an extraordinary intracellular source of mesostructured inclusions in both the cyto-and nucleoplasm of the epidermal lining of ovisacs of A. franciscana. Confocal reflection microscopy not only confirmed our finding; it also revealed reflective, light dispersing activity of these flake-like structures, their positioning and orientation with respect to the ovisac inside. Both the striation of alternating electron dense and electron-lucent components and the sharp edges of the flakes indicate self-assembly of material of yet unknown origin under supposed participation of crystallization. However, selected area electron diffraction could not verify the status of crystallization. Energy dispersive X-ray analysis measured a marked increase in nitrogen within the flake-like inclusion, and the almost complete absence of elements that are typically involved in inorganic crystallization. This rise in nitrogen could possibility be related to higher package density of proteins, achieved by mesostructure assembly. Conclusions The ovisac lining of A. franciscana is endowed with numerous mesostructured inclusions that have not been previously reported. We hypothesize that their self-assembly was from proteinaceous polycrystalline units and carbohydrates. These mesostructured flakes displayed active optical properties, as an umbrella-like, reflective cover of the ovisac, which suggests a functional role in the reproduction of A. franciscana. In turn, studies into ovisac mesostructured inclusions could help to optimizing rearing Artemia as feed for fish farming. We propose Artemia ovisacs as an in vivo model system for studying mesostructure formation. PMID:29093995

Novel mesostructured inclusions in the epidermal lining of Artemia franciscana ovisacs show optical activity.

PubMed

Hollergschwandtner, Elena; Schwaha, Thomas; Neumüller, Josef; Kaindl, Ulrich; Gruber, Daniela; Eckhard, Margret; Stöger-Pollach, Michael; Reipert, Siegfried

2017-01-01

Biomineralization, e.g., in sea urchins or mollusks, includes the assembly of mesoscopic superstructures from inorganic crystalline components and biopolymers. The resulting mesocrystals inspire biophysicists and material scientists alike, because of their extraordinary physical properties. Current efforts to replicate mesocrystal synthesis in vitro require understanding the principles of their self-assembly in vivo . One question, not addressed so far, is whether intracellular crystals of proteins can assemble with biopolymers into functional mesocrystal-like structures. During our electron microscopy studies into Artemia franciscana (Crustacea: Branchiopoda), we found initial evidence of such proteinaceous mesostructures. EM preparations with high-pressure freezing and accelerated freeze substitution revealed an extraordinary intracellular source of mesostructured inclusions in both the cyto-and nucleoplasm of the epidermal lining of ovisacs of A. franciscana . Confocal reflection microscopy not only confirmed our finding; it also revealed reflective, light dispersing activity of these flake-like structures, their positioning and orientation with respect to the ovisac inside. Both the striation of alternating electron dense and electron-lucent components and the sharp edges of the flakes indicate self-assembly of material of yet unknown origin under supposed participation of crystallization. However, selected area electron diffraction could not verify the status of crystallization. Energy dispersive X-ray analysis measured a marked increase in nitrogen within the flake-like inclusion, and the almost complete absence of elements that are typically involved in inorganic crystallization. This rise in nitrogen could possibility be related to higher package density of proteins, achieved by mesostructure assembly. The ovisac lining of A. franciscana is endowed with numerous mesostructured inclusions that have not been previously reported. We hypothesize that their self-assembly was from proteinaceous polycrystalline units and carbohydrates. These mesostructured flakes displayed active optical properties, as an umbrella-like, reflective cover of the ovisac, which suggests a functional role in the reproduction of A. franciscana . In turn, studies into ovisac mesostructured inclusions could help to optimizing rearing Artemia as feed for fish farming. We propose Artemia ovisacs as an in vivo model system for studying mesostructure formation.

[Fractographic analysis of clinically failed anterior all ceramic crowns].

PubMed

DU, Qian; Zhou, Min-bo; Zhang, Xin-ping; Zhao, Ke

2012-04-01

To identify the site of crack initiation and propagation path of clinically failed all ceramic crowns by fractographic analysis. Three clinically failed anterior IPS Empress II crowns and two anterior In-Ceram alumina crowns were retrieved. Fracture surfaces were examined using both optical stereo and scanning electron microscopy. Fractographic theory and fracture mechanics principles were applied to disclose the damage characteristics and fracture mode. All the crowns failed by cohesive failure within the veneer on the labial surface. Critical crack originated at the incisal contact area and propagated gingivally. Porosity was found within the veneer because of slurry preparation and the sintering of veneer powder. Cohesive failure within the veneer is the main failure mode of all ceramic crown. Veneer becomes vulnerable when flaws are present. To reduce the chances of chipping, multi-point occlusal contacts are recommended, and layering and sintering technique of veneering layer should also be improved.

Qualification of Ti6Al4V ELI Alloy Produced by Laser Powder Bed Fusion for Biomedical Applications

NASA Astrophysics Data System (ADS)

Yadroitsev, I.; Krakhmalev, P.; Yadroitsava, I.; Du Plessis, A.

2018-03-01

Rectangular Ti6Al4V extralow interstitials (ELI) samples were manufactured by laser powder bed fusion (LPBF) in vertical and horizontal orientations relative to the build platform and subjected to various heat treatments. Detailed analyses of porosity, microstructure, residual stress, tensile properties, fatigue, and fracture surfaces were performed based on x-ray micro-computed tomography, scanning electron microscopy, and x-ray diffraction methods. The types of fracture and the tensile fracture mechanisms of the LPBF Ti6Al4V ELI alloy were also studied. Detailed analysis of the microstructure and the corresponding mechanical properties were compared against standard specifications for conventional Ti6Al4V alloy for use in surgical implant applications. Conclusions regarding the mechanical properties and heat treatment of LPBF Ti6Al4V ELI for biomedical applications are made.

On the fractography of overload, stress corrosion, and cyclic fatigue failures in pyrolytic-carbon materials used in prosthetic heart-valve devices.

PubMed

Ritchie, R O; Dauskardt, R H; Pennisi, F J

1992-01-01

A scanning electron microscopy study is reported of the nature and morphology of fracture surfaces in pyrocarbons commonly used for the manufacture of mechanical heart-valve prostheses. Specifically, silicon-alloyed low-temperature-isotropic (LTI)-pyrolytic carbon is examined, both as a coating on graphite and as a monolithic material, following overload, stress corrosion (static fatigue), and cyclic fatigue failures in a simulated physiological environment of 37 degrees C Ringer's solution. It is found that, in contrast to most metallic materials yet in keeping with many ceramics, there are no distinct fracture morphologies in pyro-carbons which are characteristic of a specific mode of loading; fracture surfaces appear to be identical for both catastrophic and subcritical crack growth under either sustained or cyclic loading. We conclude that caution should be used in assigning the likely cause of failure of pyrolytic carbon heart-valve components using fractographic examination.

Non-ionic surfactant vesicles in pulmonary glucocorticoid delivery: characterization and interaction with human lung fibroblasts.

PubMed

Marianecci, Carlotta; Paolino, Donatella; Celia, Christian; Fresta, Massimo; Carafa, Maria; Alhaique, Franco

2010-10-01

Non-ionic surfactant vesicles (NSVs) were proposed for the pulmonary delivery of glucocorticoids such as beclomethasone dipropionate (BDP) for the treatment of inflammatory lung diseases, e.g. asthma, chronic obstructive pulmonary disease and various type of pulmonary fibrosis. The thin layer evaporation method followed by sonication was used to prepare small non-ionic surfactant vesicles containing beclomethasone dipropionate. Light scattering experiments showed that beclomethasone dipropionate-loaded non-ionic surfactant vesicles were larger than unloaded ones and showed a significant (P<0.001) decrease of the zeta potential. The morphological analysis, by freeze-fracture transmission electron microscopy, showed the maintenance of a vesicular structure in the presence of the drug. The colloidal and storage stability were evaluated by Turbiscan Lab Expert, which evidenced the good stability of BDP-loaded non-ionic surfactant vesicles, thus showing no significant variations of mean size and no colloidal phase segregation. Primary human lung fibroblast (HLF) cells were used for in vitro investigation of vesicle tolerability, carrier-cell interaction, intracellular drug uptake and drug-loaded vesicle anti-inflammatory activity. The investigated NSVs did not show a significant cytotoxic activity at all incubation times for concentrations ranging from 0.01 to 1 μM. Confocal laser scanning microscopy showed vesicular carrier localization at the level of the cytoplasm compartment, where the glucocorticoid receptor (target site) is localized. BDP-loaded non-ionic surfactant vesicles elicited a significant improvement of the HLF intracellular uptake of the drug with respect to the free drug solution, drug/surfactant mixtures and empty vesicles used as references. The treatment of HLF cells with BDP-loaded non-ionic surfactant vesicles determined a noticeable increase of the drug anti-inflammatory activity by reducing the secretion of both constitutive and interleukin-1β-stimulated nerve growth factor (as inflammatory index) of 68% and 85%, respectively. Obtained data indicate that the investigated NSVs represent a promising tool as a pulmonary drug delivery system. Copyright © 2010 Elsevier B.V. All rights reserved.

Thermodynamically stable vesicle formation from glycolipid biosurfactant sponge phase.

PubMed

Imura, Tomohiro; Yanagishita, Hiroshi; Ohira, Junko; Sakai, Hideki; Abe, Masahiko; Kitamoto, Dai

2005-06-25

Thermodynamically stable vesicle (L(alpha1)) formation from glycolipid biosurfactant sponge phase (L(3)) and its mechanism were investigated using a "natural" biocompatible mannosyl-erythritol lipid-A (MEL-A)/L-alpha-dilauroylphosphatidylcholine (DLPC) mixture by varying the composition. The trapping efficiency for calcein and turbidity measurements clearly indicated the existence of three regions: while the trapping efficiencies of the mixed MEL-A/DLPC assemblies at the compositions with X(DLPC)< or =0.1 or X(DLPC)> or =0.8 were almost zero, the mixed assemblies at the compositions with 0.1 or =0.8 were multilamellar vesicles (L(alpha)) with diameter from 2 to 10 microm. Meanwhile, dynamic light scattering (DLS) measurement revealed that the average size of the vesicles at the composition of X(DLPC)=0.3 was 633.2 nm, which is remarkably small compared to other compositions. Moreover, the mixed vesicle solution at the composition of X(DLPC)=0.3 was slightly bluish and turbid and kept its dispersion stability at 25 degrees C for more than 3 months, indicating the formation of a thermodynamically stable vesicle (L(alpha1)). These results exhibited the formation of a thermodynamically stable vesicle (L(alpha1)) with a high dispersibility from the MEL-A/DLPC mixture. The asymmetric distribution of MEL-A and DLPC in the two vesicle monolayers caused by the difference in geometrical structures is very likely to have changed their self-assembled structure from a sponge phase (L(3)) to a thermodynamically stable vesicle (L(alpha1)).

Morphology and Dynamic Mechanical Properties of Diglycidyl Ether of Bisphenol-A Toughened with Carboxyl-Terminated Butadiene-Acrylonitrile

NASA Technical Reports Server (NTRS)

Hong, S. D.; Chung, S. Y.; Fedors, R. F.; Moacanin, J.; Gupta, A.

1984-01-01

The fracture toughness of an incorporation of a carboxyl-terminated butadiene acrylonitrile (CTBN) elastomer in diglycidyl ether bisphenol A (DGEBA) resin was investigated. Measurements of dynamic mechanical properties, scanning electron microscopy and small-angle X-ray scattering were carried out to characterize the state of cure, morphology and particle size and size distribution of the neat resins and their graphite fiber reinforced composites.

Effect of LID (Registered) processing on the microstructure and mechanical properties of Ti-6Al-4V and Ti-6Al-2Sn-4Zr-2Mo titanium foil-gauge materials

NASA Technical Reports Server (NTRS)

Balckburn, Linda B.

1987-01-01

A study was undertaken to determine the mechanical properties and microstructures resulting from Liquid Interface Diffusion (LID -Registered) processing of foil-gauge specimens of Ti-6Al-4V and Ti-6Al-2Sn-4Zr-2Mo coated with varying amounts of LID material. In addition, the effects of various elevated temperature exposures on the concentration profiles of the LID alloying elements were investigated, using specimens with a narrow strip of LID material applied to the surface. Room and elevated temperature tensile properties were determined for both coated and uncoated specimens. Optical microscopy was used to examine alloy microstructures, and scanning electron microscopy to examine fracture surface morphologies. The chemical concentration profiles of the strip-coated specimens were determined with an electron microprobe.

Microstructure and Interfacial Shear Strength in W/(Zr55Cu30Al10Ni5)100- x Nb x Composites

NASA Astrophysics Data System (ADS)

Mahmoodan, M.; Gholamipour, R.; Mirdamadi, Sh.; Nategh, S.

2017-11-01

In the present study, (Zr55Cu30Al10Ni5)100- x Nb( x=0,1,2,3) bulk metallic glass matrix/tungsten wire composites were fabricated by a gas pressure infiltration process at temperature 950 °C for 5 min. Microstructural studies and mechanical behaviors of the materials have been investigated by scanning electron microscopy, transmission electron microscopy and pullout tests. The mechanical results showed that the interface shear strength in the composite sample with X = 2 increased more than twice compared to the composite sample with X = 0. Based on the microstructural results, the addition of two atomic percent Nb in the matrix composite causes an increase in the diffusion band thickness during the melt infiltration and change in the interface fracture mode as a result of pullout test.

Scanning Electron Microscopy Investigation of a Sample Depth Profile Through the Martian Meteorite Nakhla

NASA Technical Reports Server (NTRS)

Toporski, Jan; Steele, Andrew; Westall, Frances; McKay, David S.

2000-01-01

The ongoing scientific debate as to whether or not the Martian meteorite ALH84001 contained evidence of possible biogenic activities showed the need to establish consistent methods to ascertain the origin of such evidence. To distinguish between terrestrial organic material/microbial contaminants and possible indigenous microbiota within meteorites is therefore crucial. With this in mind a depth profile consisting of four samples from a new sample allocation of Martian meteorite Nakhla was investigated using scanning electron microscopy (SEM) and energy dispersive X-ray analysis. SEM imaging of freshly broken fractured chips revealed structures strongly recent terrestrial microorganisms, in some cases showing evidence of active growth. This conclusion was supported by EDX analysis, which showed the presence of carbon associated with these structures, we concluded that these structures represent recent terrestrial contaminants rather than structures indigenous to the meteorite. Page

Comparison of mechanical characteristics of focused ion beam fabricated silicon nanowires

NASA Astrophysics Data System (ADS)

Ina, Ginnosuke; Fujii, Tatsuya; Kozeki, Takahiro; Miura, Eri; Inoue, Shozo; Namazu, Takahiro

2017-06-01

In this study, we investigate the effects of focused ion beam (FIB)-induced damage and specimen size on the mechanical properties of Si nanowires (NWs) by a microelectromechanical system (MEMS)-based tensile testing technique. By an FIB fabrication technique, three types of Si NWs, which are as-FIB-fabricated, annealed, and FIB-implanted NWs, are prepared. A sacrificial-oxidized NW is also prepared to compare the mechanical properties of these FIB-based NWs. The quasi-static uniaxial tensile tests of all the NWs are conducted by scanning electron microscopy (SEM). The fabrication process and specimen size dependences on Young’s modulus and fracture strength are observed. Annealing is effective for improving the Young’s modulus of the FIB-damaged Si. Transmission electron microscopy (TEM) suggests that the mechanism behind the process dependence on the mechanical characteristics is related to the crystallinity of the FIB-damaged portion.

Predictions and Experimental Microstructural Characterization of High Strain Rate Failure Modes in Layered Aluminum Composites

NASA Astrophysics Data System (ADS)

Khanikar, Prasenjit

Different aluminum alloys can be combined, as composites, for tailored dynamic applications. Most investigations pertaining to metallic alloy layered composites, however, have been based on quasi-static approaches. The dynamic failure of layered metallic composites, therefore, needs to be characterized in terms of strength, toughness, and fracture response. A dislocation-density based crystalline plasticity formulation, finite-element techniques, rational crystallographic orientation relations and a new fracture methodology were used to predict the failure modes associated with the high strain rate behavior of aluminum layered composites. Two alloy layers, a high strength alloy, aluminum 2195, and an aluminum alloy 2139, with high toughness, were modeled with representative microstructures that included precipitates, dispersed particles, and different grain boundary (GB) distributions. The new fracture methodology, based on an overlap method and phantom nodes, is used with a fracture criteria specialized for fracture on different cleavage planes. One of the objectives of this investigation, therefore, was to determine the optimal arrangements of the 2139 and 2195 aluminum alloys for a metallic layered composite that would combine strength, toughness and fracture resistance for high strain-rate applications. Different layer arrangements were investigated for high strain-rate applications, and the optimal arrangement was with the high toughness 2139 layer on the bottom, which provided extensive shear strain localization, and the high strength 2195 layer on the top for high strength resistance. The layer thickness of the bottom high toughness layer also affected the bending behavior of the roll-boned interface and the potential delamination of the layers. Shear strain localization, dynamic cracking and delamination were the mutually competing failure mechanisms for the layered metallic composite, and control of these failure modes can be optimized for high strain-rate applications. The second major objective of this investigation was the use of recently developed dynamic fracture formulations to model and analyze the crack nucleation and propagation of aluminum layered composites subjected to high strain rate loading conditions and how microstructural effects, such as precipitates, dispersed particles, and GB orientations affect failure evolution. This dynamic fracture approach is used to investigate crack nucleation and crack growth as a function of the different microstructural characteristics of each alloy in layered composites with and without pre-existing cracks. The zigzag nature of the crack paths were mainly due to the microstructural features, such as precipitates and dispersed particles distributions and orientations ahead of the crack front, and it underscored the capabilities of the fracture methodology. The evolution of dislocation density and the formation of localized shear slip contributed to the blunting of the propagating crack. Extensive geometrical and thermal softening due to the localized plastic slip also affected crack path orientations and directions. These softening mechanisms resulted in the switching of cleavage planes, which affected crack path orientations. Interface delamination can also have an important role in the failure and toughening of the layered composites. Different scenarios of delamination were investigated, such as planar crack growth and crack penetration into the layers. The presence of brittle surface oxide platelets in the interface region also significantly influenced the interface delamination process. Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM) and Optical Microscopy (OM) characterization provided further physical insights and validation of the predictive capabilities. The inherent microstructural features of each alloy play a significant role in the dynamic fracture, shear strain localization, and interface delamination of the layered metallic composite. These microstructural features, such as precipitates, dispersed particles, and GB orientations and distributions can be optimized for desired behavior of metallic composites.

High-pressure freezing for scanning transmission electron tomography analysis of cellular organelles.

PubMed

Walther, Paul; Schmid, Eberhard; Höhn, Katharina

2013-01-01

Using an electron microscope's scanning transmission mode (STEM) for collection of tomographic datasets is advantageous compared to bright field transmission electron microscopic (TEM). For image formation, inelastic scattering does not cause chromatic aberration, since in STEM mode no image forming lenses are used after the beam has passed the sample, in contrast to regular TEM. Therefore, thicker samples can be imaged. It has been experimentally demonstrated that STEM is superior to TEM and energy filtered TEM for tomography of samples as thick as 1 μm. Even when using the best electron microscope, adequate sample preparation is the key for interpretable results. We adapted protocols for high-pressure freezing of cultivated cells from a physiological state. In this chapter, we describe optimized high-pressure freezing and freeze substitution protocols for STEM tomography in order to obtain high membrane contrast.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Rubing, E-mail: zrb86411680@126.com; Zhang, Yaoyao; Liu, Qiang

TiAl/Nb and TiAl/NiCoCrAl laminate composite sheets with a thickness of 0.4–0.6 mm and dimensions of 150 mm × 100 mm were successfully fabricated by electron beam physical vapor deposition. The microstructures of the sheets were examined, and their mechanical properties were compared with those of TiAl monolithic sheet produced by electron beam physical vapor deposition. Tensile testing was performed at room temperature and 750 °C, and the fracture surfaces were examined by scanning electron microscopy. Among the three microlaminate sheets, the TiAl/NiCoCrAl micro-laminate sheet had the best comprehensive properties at room temperature, and the TiAl/Nb micro-laminate sheet showed the idealmore » high-temperature strength and plasticity at 750 °C. The result was discussed in terms of metal strengthening mechanism. - Highlights: • TiAl-based multilayer foils was fabricated successfully by using EB-PVD method; • The tensile properties and micro-fracture morphologies of the sheet were investigated; • The deformation behavior of the multilayer foils was discussed.« less

Chrysophyte cysts as potential environmental indicators.

USGS Publications Warehouse

Adam, D.P.; Mahood, A.D.

1981-01-01

Many chrysophyte algae produce morphologically distinctive, siliceous, microscopic cysts during a resting stage of their life cycles; these cysts are often preserved in sediments. Scanning electron microscopy and Nomarski optics permit much more detailed observation of these cysts than was heretofore possible. Many cyst types are found only in specific habitats, such as montane lakes, wet meadows, ephemeral ponds, and Sphagnum bogs. In the samples studied, cysts seem to be most common in fluctuating fresh-water habitats of low to moderate pH and some winter freezing. Chrysophyte cysts have the potential to be a useful tool for modern environmental assessments and paleoecological studies of Cenozoic fresh-water lacustrine deposits. -from Authors

Cryo-STEM-EDX spectroscopy for the characterisation of nanoparticles in cell culture media

NASA Astrophysics Data System (ADS)

Ilett, M.; Bamiduro, F.; Matar, O.; Brown, A.; Brydson, R.; Hondow, N.

2017-09-01

We present a study of barium titanate nanoparticles dispersed in cell culture media. Scanning transmission electron microscopy combined with energy dispersive X-ray spectroscopy was undertaken on samples prepared using both conventional drop casting and also plunge freezing and examination under cryogenic conditions. This showed that drying artefacts occurred during conventional sample preparation, whereby some salt components of the cell culture media accumulated around the barium titanate nanoparticles; these were removed using the cryogenic route. Importantly, the formation of a calcium and phosphorus rich coating around the barium titanate nanoparticles was retained under cryo-conditions, highlighting that significant interactions do occur between nanomaterials and biological media.

Bioinspired Hydroxyapatite/Poly(methyl methacrylate) Composite with a Nacre-Mimetic Architecture by a Bidirectional Freezing Method.

PubMed

Bai, Hao; Walsh, Flynn; Gludovatz, Bernd; Delattre, Benjamin; Huang, Caili; Chen, Yuan; Tomsia, Antoni P; Ritchie, Robert O

2016-01-06

Using a bidirectional freezing technique, combined with uniaxial pressing and in situ polymerization, "nacre-mimetic" hydroxyapatite/poly(methyl methacrylate) (PMMA) composites are developed by processing large-scale aligned lamellar ceramic scaffolds. Structural and mechanical characterization shows "brick-and-mortar" structures, akin to nacre, with interesting combinations of strength, stiffness, and work of fracture, which provide a pathway to making strong and tough lightweight materials. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Application of Optical Coherence Tomography Freeze-Drying Microscopy for Designing Lyophilization Process and Its Impact on Process Efficiency and Product Quality.

PubMed

Korang-Yeboah, Maxwell; Srinivasan, Charudharshini; Siddiqui, Akhtar; Awotwe-Otoo, David; Cruz, Celia N; Muhammad, Ashraf

2018-01-01

Optical coherence tomography freeze-drying microscopy (OCT-FDM) is a novel technique that allows the three-dimensional imaging of a drug product during the entire lyophilization process. OCT-FDM consists of a single-vial freeze dryer (SVFD) affixed with an optical coherence tomography (OCT) imaging system. Unlike the conventional techniques, such as modulated differential scanning calorimetry (mDSC) and light transmission freeze-drying microscopy, used for predicting the product collapse temperature (Tc), the OCT-FDM approach seeks to mimic the actual product and process conditions during the lyophilization process. However, there is limited understanding on the application of this emerging technique to the design of the lyophilization process. In this study, we investigated the suitability of OCT-FDM technique in designing a lyophilization process. Moreover, we compared the product quality attributes of the resulting lyophilized product manufactured using Tc, a critical process control parameter, as determined by OCT-FDM versus as estimated by mDSC. OCT-FDM analysis revealed the absence of collapse even for the low protein concentration (5 mg/ml) and low solid content formulation (1%w/v) studied. This was confirmed by lab scale lyophilization. In addition, lyophilization cycles designed using Tc values obtained from OCT-FDM were more efficient with higher sublimation rate and mass flux than the conventional cycles, since drying was conducted at higher shelf temperature. Finally, the quality attributes of the products lyophilized using Tc determined by OCT-FDM and mDSC were similar, and product shrinkage and cracks were observed in all the batches of freeze-dried products irrespective of the technique employed in predicting Tc.

Storing Blood Cells

NASA Technical Reports Server (NTRS)

1976-01-01

The National Cancer Institute worked with Goddard Space Flight Center to propose a solution to the blood-cell freezing problem. White blood cells and bone marrow are stored for future use by leukemia patients as a result of Goddard and Jet Propulsion Laboratory expertise in electronics and cryogenics. White blood cell and bone marrow bank established using freezing unit. Freezing unit monitors temperature of cells themselves. Thermocouple placed against polyethylene container relays temperature signals to an electronic system which controls small heaters located outside container. Heaters allow liquid nitrogen to circulate at constant temperature and maintain consistent freezing rate. Ability to freeze, store, and thaw white cells and bone marrow without damage is important in leukemia treatment.

A study of fractography in the low-temperature brittle fracture of an 18Cr-18Mn-0.7N austenitic steel

DOE Office of Scientific and Technical Information (OSTI.GOV)

Liu, S.C.; Hashida, T.; Takahashi, H.

1998-03-01

The fracture mode and crack propagation behavior of brittle fracture at 77 and 4 K in an 18Cr-18Mn-0.7N austenitic stainless were investigated using optical and scanning electron microscopy. The fracture path was examined by observing the side surface in a partially ruptured specimen. The relationship of the fracture facets to the microstructures were established by observing the fracture surface and the adjacent side surface simultaneously. Three kinds of fracture facets were identified at either temperature. The first is a smooth curved intergranular fracture facet with characteristic parallel lines on it. The second is a fairly planar facet formed by partingmore » along an annealing twin boundary, a real {l_brace}111{r_brace} plane. There are three sets of parallel lines on the facet and the lines in different sets intersect at 60 deg. The third is a lamellar transgranular fracture facet with sets of parallel steps on it. Fracture propagated by the formation of microcracks on a grain boundary, annealing twin boundary, and coalescence of these cracks. The observation suggests that the ease of crack initiation and propagation along the grain boundary and the annealing twin boundary may be the main reason for the low-temperature brittleness of this steel. A mechanism for grain boundary cracking, including annealing twin boundary parting, has been discussed based on the stress concentration induced by impinging planar deformation structures on the grain boundaries.« less

Dissolution-Driven Permeability Reduction of a Fractured Carbonate Caprock

PubMed Central

Ellis, Brian R.; Fitts, Jeffrey P.; Bromhal, Grant S.; McIntyre, Dustin L.; Tappero, Ryan; Peters, Catherine A.

2013-01-01

Abstract Geochemical reactions may alter the permeability of leakage pathways in caprocks, which serve a critical role in confining CO2 in geologic carbon sequestration. A caprock specimen from a carbonate formation in the Michigan sedimentary Basin was fractured and studied in a high-pressure core flow experiment. Inflowing brine was saturated with CO2 at 40°C and 10 MPa, resulting in an initial pH of 4.6, and had a calcite saturation index of −0.8. Fracture permeability decreased during the experiment, but subsequent analyses did not reveal calcite precipitation. Instead, experimental observations indicate that calcite dissolution along the fracture pathway led to mobilization of less soluble mineral particles that clogged the flow path. Analyses of core sections via electron microscopy, synchrotron-based X-ray diffraction imaging, and the first application of microbeam Ca K-edge X-ray absorption near edge structure, provided evidence that these occlusions were fragments from the host rock rather than secondary precipitates. X-ray computed tomography showed a significant loss of rock mass within preferential flow paths, suggesting that dissolution also removed critical asperities and caused mechanical closure of the fracture. The decrease in fracture permeability despite a net removal of material along the fracture pathway demonstrates a nonintuitive, inverse relationship between dissolution and permeability evolution in a fractured carbonate caprock. PMID:23633894

Dynamic and cyclic fatigue of engine-driven rotary nickel-titanium endodontic instruments.

PubMed

Haïkel, Y; Serfaty, R; Bateman, G; Senger, B; Allemann, C

1999-06-01

The absence of adequate testing standards for engine-driven nickel-titanium (NiTi) instruments necessitates further study of these instruments in all areas. This study examined three groups of engine-driven rotary NiTi endodontic instruments (Profile, Hero, and Quantec) and assessed the times for dynamic fracture in relation to the radius of curvature to which the instruments were subjected during preparation, with the instrument diameter determined by size and taper and the mode by which the fracture occurred. Ten instruments were randomly selected representing each size and taper for each group and for each radius of curvature: 600 in total. The instruments were rotated at 350 rpm and introduced into a tempered steel curve that simulated a canal. Two radii of curvature of canals were used: 5 and 10 mm. Time at fracture was noted for all files, and the fracture faces of each file were analyzed with scanning electron microscopy. Radius of curvature was found to be the most significant factor in determining the fatigue resistance of the files. As radius of curvature decreased, fracture time decreased. Taper of files was found to be significant in determining fracture time. As diameter increased, fracture time decreased. In all cases, fracture was found to be of a ductile nature, thus implicating cyclic fatigue as a major cause of failure and necessitating further analyses and setting of standards in this area.

Cleavage fracture in high strength low alloy weld metal

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bose, W.W.; Bowen, P.; Strangwood, M.

1996-12-31

The present investigation gives an evaluation of the effect of microstructure on the cleavage fracture process of High Strength Low Alloy (HSLA) multipass weld metals. With additions of alloying elements, such as Ti, Ni, Mo and Cr, the microstructure of C-Mn weld metal changes from the classical composition, i.e., allotriomorphic ferrite with acicular ferrite and Widmanstaetten ferrite, to bainite and low carbon martensite. Although the physical metallurgy of some HSLA weld metals has been studied before, more work is necessary to correlate the effect of the microstructure on the fracture behavior of such weld metals. In this work detailed microstructuralmore » analysis was carried out using optical and electron (SEM and TEM) microscopy. Single edge notched (SEN) bend testpieces were used to assess the cleavage fracture stress, {sigma}{sub F}. Inclusions beneath the notch surface were identified as the crack initiators of unstable cleavage fracture. From the size of such inclusions and the value of tensile stress predicted at the initiation site, the effective surface energy for cleavage was calculated using a modified Griffth energy balance for a penny shape crack. The results suggest that even though inclusions initiate cleavage fracture, the local microstructure may play an important role in the fracture process of these weld metals. The implications of these observations for a quantitative theory of the cleavage fracture of ferritic steels is discussed.« less

The effect of residual stresses induced by prestraining on fatigue life of notched specimens

NASA Astrophysics Data System (ADS)

Sadeler, R.; Ozel, A.; Kaymaz, I.; Totik, Y.

2005-06-01

The effect of tensile prestraining-induced residual stress on the fatigue life of notched steel parts was investigated. The study was performed on AISI 4140 steel. Rotating bending fatigue tests were carried out on semicircular notched specimens with different notch radii in the as-quenched and tempered conditions. Metallography of the specimens was performed by means of light optical microscopy. The finite-element method was used to evaluate the residual stress distribution near the notch region. Fatigue tests revealed fatigue life improvement for notched specimens, which changes depending on the notch radii and applied stress. Scanning electron microscopy was used to examine the fracture surfaces of the specimens.

Characterization of multiwalled carbon nanotube-polymethyl methacrylate composite resins as denture base materials.

PubMed

Wang, Russell; Tao, Junliang; Yu, Bill; Dai, Liming

2014-04-01

Most fractures of dentures occur during function, primarily because of the flexural fatigue of denture resins. The purpose of this study was to evaluate a polymethyl methacrylate denture base material modified with multiwalled carbon nanotubes in terms of fatigue resistance, flexural strength, and resilience. Denture resin specimens were fabricated: control, 0.5 wt%, 1 wt%, and 2 wt% of multiwalled carbon nanotubes. Multiwalled carbon nanotubes were dispersed by sonication. Thermogravimetric analysis was used to determine quantitative dispersions of multiwalled carbon nanotubes in polymethyl methacrylate. Raman spectroscopic analyses were used to evaluate interfacial reactions between the multiwalled carbon nanotubes and the polymethyl methacrylate matrix. Groups with and without multiwalled carbon nanotubes were subjected to a 3-point-bending test for flexural strength. Resilience was derived from a stress and/or strain curve. Fatigue resistance was conducted by a 4-point bending test. Fractured surfaces were analyzed by scanning electron microscopy. One-way ANOVA and the Duncan tests were used to identify any statistical differences (α=.05). Thermogravimetric analysis verified the accurate amounts of multiwalled carbon nanotubes dispersed in the polymethyl methacrylate resin. Raman spectroscopy showed an interfacial reaction between the multiwalled carbon nanotubes and the polymethyl methacrylate matrix. Statistical analyses revealed significant differences in static and dynamic loadings among the groups. The worst mechanical properties were in the 2 wt% multiwalled carbon nanotubes (P<.05), and 0.5 wt% and 1 wt% multiwalled carbon nanotubes significantly improved flexural strength and resilience. All multiwalled carbon nanotubes-polymethyl methacrylate groups showed poor fatigue resistance. The scanning electron microscopy results indicated more agglomerations in the 2% multiwalled carbon nanotubes. Multiwalled carbon nanotubes-polymethyl methacrylate groups (0.5% and 1%) performed better than the control group during the static flexural test. The results indicated that 2 wt% multiwalled carbon nanotubes were not beneficial because of the inadequate dispersion of multiwalled carbon nanotubes in the polymethyl methacrylate matrix. Scanning electron microscopy analysis showed agglomerations on the fracture surface of 2 wt% multiwalled carbon nanotubes. The interfacial bonding between multiwalled carbon nanotubes and polymethyl methacrylate was weak based on the Raman data and dynamic loading results. Copyright © 2014 Editorial Council for the Journal of Prosthetic Dentistry. Published by Elsevier Inc. All rights reserved.

Enhancement of Apoptosis by Titanium Alloy Internal Fixations during Microwave Treatments for Fractures: An Animal Study.

PubMed

Wang, Gang; Xu, Yiming; Zhang, Lina; Ye, Dongmei; Feng, Xianxuan; Fu, Tengfei; Bai, Yuehong

2015-01-01

Microwaves are used in one method of physical therapy and can increase muscle tissue temperature which is useful for improving muscle, tendon and bone injuries. In the study, we sought to determine whether titanium alloy internal fixations influence apoptosis in tissues subjected to microwave treatments at 2,450 MHz and 40 W during the healing of fractures because this issue is not yet fully understood. In this study, titanium alloy internal fixations were used to treat 3.0-mm transverse osteotomies in the middle of New Zealand rabbits' femurs. After the operation, 30-day microwave treatments were applied to the 3.0 mm transverse osteotomies 3 days after the operation. The changes in the temperatures of the muscle tissues in front of the implants or the 3.0 mm transverse osteotomies were measured during the microwave treatments. To characterize the effects of titanium alloy internal fixations on apoptosis in the muscles after microwave treatment, we performed TUNEL assays, fluorescent real-time (quantitative) PCR, western blotting analyses, reactive oxygen species (ROS) detection and transmission electron microscopy examinations. The temperatures were markedly increased in the animals with the titanium alloy implants. Apoptosis in the muscle cells of the implanted group was significantly more extensive than that in the non-implanted control group at different time points. Transmission electron microscopy examinations of the skeletal muscles of the implanted groups revealed muscular mitochondrial swelling, vacuolization. ROS, Bax and Hsp70 were up-regulated, and Bcl-2 was down-regulated in the implanted group. Our results suggest that titanium alloy internal fixations caused greater muscular tissue cell apoptosis following 2,450 MHz, 40 W microwave treatments in this rabbit femur fracture models.

Enhancement of Apoptosis by Titanium Alloy Internal Fixations during Microwave Treatments for Fractures: An Animal Study

PubMed Central

Zhang, Lina; Ye, Dongmei; Feng, Xianxuan; Fu, Tengfei; Bai, Yuehong

2015-01-01

Objective Microwaves are used in one method of physical therapy and can increase muscle tissue temperature which is useful for improving muscle, tendon and bone injuries. In the study, we sought to determine whether titanium alloy internal fixations influence apoptosis in tissues subjected to microwave treatments at 2,450 MHz and 40 W during the healing of fractures because this issue is not yet fully understood. Methods In this study, titanium alloy internal fixations were used to treat 3.0-mm transverse osteotomies in the middle of New Zealand rabbits’ femurs. After the operation, 30-day microwave treatments were applied to the 3.0 mm transverse osteotomies 3 days after the operation. The changes in the temperatures of the muscle tissues in front of the implants or the 3.0 mm transverse osteotomies were measured during the microwave treatments. To characterize the effects of titanium alloy internal fixations on apoptosis in the muscles after microwave treatment, we performed TUNEL assays, fluorescent real-time (quantitative) PCR, western blotting analyses, reactive oxygen species (ROS) detection and transmission electron microscopy examinations. Results The temperatures were markedly increased in the animals with the titanium alloy implants. Apoptosis in the muscle cells of the implanted group was significantly more extensive than that in the non-implanted control group at different time points. Transmission electron microscopy examinations of the skeletal muscles of the implanted groups revealed muscular mitochondrial swelling, vacuolization. ROS, Bax and Hsp70 were up-regulated, and Bcl-2 was down-regulated in the implanted group. Conclusion Our results suggest that titanium alloy internal fixations caused greater muscular tissue cell apoptosis following 2,450 MHz, 40 W microwave treatments in this rabbit femur fracture models. PMID:26132082

Do's and don'ts of cryo-electron microscopy: a primer on sample preparation and high quality data collection for macromolecular 3D reconstruction.

PubMed

Cabra, Vanessa; Samsó, Montserrat

2015-01-09

Cryo-electron microscopy (cryoEM) entails flash-freezing a thin layer of sample on a support, and then visualizing the sample in its frozen hydrated state by transmission electron microscopy (TEM). This can be achieved with very low quantity of protein and in the buffer of choice, without the use of any stain, which is very useful to determine structure-function correlations of macromolecules. When combined with single-particle image processing, the technique has found widespread usefulness for 3D structural determination of purified macromolecules. The protocol presented here explains how to perform cryoEM and examines the causes of most commonly encountered problems for rational troubleshooting; following all these steps should lead to acquisition of high quality cryoEM images. The technique requires access to the electron microscope instrument and to a vitrification device. Knowledge of the 3D reconstruction concepts and software is also needed for computerized image processing. Importantly, high quality results depend on finding the right purification conditions leading to a uniform population of structurally intact macromolecules. The ability of cryoEM to visualize macromolecules combined with the versatility of single particle image processing has proven very successful for structural determination of large proteins and macromolecular machines in their near-native state, identification of their multiple components by 3D difference mapping, and creation of pseudo-atomic structures by docking of x-ray structures. The relentless development of cryoEM instrumentation and image processing techniques for the last 30 years has resulted in the possibility to generate de novo 3D reconstructions at atomic resolution level.

Notched graphite polymimide composites at room and notched graphite polymide composites at room and elevated temperatures. [nondestructive test techniques

NASA Technical Reports Server (NTRS)

Awerbuch, J.; Perkinson, H. E.; Kamel, I. L.

1980-01-01

The fracture behavior in graphite/polyimide (Gr/PI) Celion 6000/PMR-15 composites was characterized. Emphasis was placed on the correlation between the observed failure modes and the deformation characteristics of center-notched Gr/Pl laminates. Crack tip damage growth, fracture strength and notch sensitivity, and the associated characterization methods were also examined. Special attention was given to nondestructive evaluation of internal damage and damage growth, techniques such as acoustic emission, X-ray radiography, and ultrasonic C-scan. Microstructural studies using scanning electron microscopy, photomicrography, and the pulsed nuclear magnetic resonance technique were employed as well. All experimental procedures and techniques are described and a summary of representative results for Gr/Pl laminates is given.

Comparison of the fracture resistances of glass fiber mesh- and metal mesh-reinforced maxillary complete denture under dynamic fatigue loading

PubMed Central

2017-01-01

PURPOSE The aim of this study was to investigate the effect of reinforcing materials on the fracture resistances of glass fiber mesh- and Cr–Co metal mesh-reinforced maxillary complete dentures under fatigue loading. MATERIALS AND METHODS Glass fiber mesh- and Cr–Co mesh-reinforced maxillary complete dentures were fabricated using silicone molds and acrylic resin. A control group was prepared with no reinforcement (n = 15 per group). After fatigue loading was applied using a chewing simulator, fracture resistance was measured by a universal testing machine. The fracture patterns were analyzed and the fractured surfaces were observed by scanning electron microscopy. RESULTS After cyclic loading, none of the dentures showed cracks or fractures. During fracture resistance testing, all unreinforced dentures experienced complete fracture. The mesh-reinforced dentures primarily showed posterior framework fracture. Deformation of the all-metal framework caused the metal mesh-reinforced denture to exhibit the highest fracture resistance, followed by the glass fiber mesh-reinforced denture (P<.05) and the control group (P<.05). The glass fiber mesh-reinforced denture primarily maintained its original shape with unbroken fibers. River line pattern of the control group, dimples and interdendritic fractures of the metal mesh group, and radial fracture lines of the glass fiber group were observed on the fractured surfaces. CONCLUSION The glass fiber mesh-reinforced denture exhibits a fracture resistance higher than that of the unreinforced denture, but lower than that of the metal mesh-reinforced denture because of the deformation of the metal mesh. The glass fiber mesh-reinforced denture maintains its shape even after fracture, indicating the possibility of easier repair. PMID:28243388

Assessment of different sample preparation routes for mass spectrometric monitoring and imaging of lipids in bone cells via ToF-SIMS

PubMed Central

Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Wenisch, Sabine; Janek, Jürgen

2015-01-01

In ToF-SIMS analysis, the experimental outcome from cell experiments is to a great extent influenced by the sample preparation routine. In order to better judge this critical influence in the case of lipid analysis, a detailed comparison of different sample preparation routines is performed—aiming at an optimized preparation routine for systematic lipid imaging of cell cultures. For this purpose, human mesenchymal stem cells were analyzed: (a) as chemically fixed, (b) freeze-dried, and (c) frozen-hydrated. For chemical fixation, different fixatives, i.e., glutaraldehyde, paraformaldehyde, and a mixture of both, were tested with different postfixative handling procedures like storage in phosphate buffered saline, water or critical point drying. Furthermore, secondary lipid fixation via osmium tetroxide was taken into account and the effect of an ascending alcohol series with and without this secondary lipid fixation was evaluated. Concerning freeze-drying, three different postprocessing possibilities were examined. One can be considered as a pure cryofixation technique while the other two routes were based on chemical fixation. Cryofixation methods known from literature, i.e., freeze-fracturing and simple frozen-hydrated preparation, were also evaluated to complete the comparison of sample preparation techniques. Subsequent data evaluation of SIMS spectra in both, positive and negative, ion mode was performed via principal component analysis by use of peak sets representative for lipids. For freeze-fracturing, these experiments revealed poor reproducibility making this preparation route unsuitable for systematic investigations and statistic data evaluation. Freeze-drying after cryofixation showed improved reproducibility and well preserved lipid contents while the other freeze-drying procedures showed drawbacks in one of these criteria. In comparison, chemical fixation techniques via glutar- and/or paraformaldehyde proved most suitable in terms of reproducibility and preserved lipid contents, while alcohol and osmium treatment led to the extraction of lipids and are therefore not recommended. PMID:25791294

Assessment of different sample preparation routes for mass spectrometric monitoring and imaging of lipids in bone cells via ToF-SIMS.

PubMed

Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Wenisch, Sabine; Janek, Jürgen

2015-03-19

In ToF-SIMS analysis, the experimental outcome from cell experiments is to a great extent influenced by the sample preparation routine. In order to better judge this critical influence in the case of lipid analysis, a detailed comparison of different sample preparation routines is performed-aiming at an optimized preparation routine for systematic lipid imaging of cell cultures. For this purpose, human mesenchymal stem cells were analyzed: (a) as chemically fixed, (b) freeze-dried, and (c) frozen-hydrated. For chemical fixation, different fixatives, i.e., glutaraldehyde, paraformaldehyde, and a mixture of both, were tested with different postfixative handling procedures like storage in phosphate buffered saline, water or critical point drying. Furthermore, secondary lipid fixation via osmium tetroxide was taken into account and the effect of an ascending alcohol series with and without this secondary lipid fixation was evaluated. Concerning freeze-drying, three different postprocessing possibilities were examined. One can be considered as a pure cryofixation technique while the other two routes were based on chemical fixation. Cryofixation methods known from literature, i.e., freeze-fracturing and simple frozen-hydrated preparation, were also evaluated to complete the comparison of sample preparation techniques. Subsequent data evaluation of SIMS spectra in both, positive and negative, ion mode was performed via principal component analysis by use of peak sets representative for lipids. For freeze-fracturing, these experiments revealed poor reproducibility making this preparation route unsuitable for systematic investigations and statistic data evaluation. Freeze-drying after cryofixation showed improved reproducibility and well preserved lipid contents while the other freeze-drying procedures showed drawbacks in one of these criteria. In comparison, chemical fixation techniques via glutar- and/or paraformaldehyde proved most suitable in terms of reproducibility and preserved lipid contents, while alcohol and osmium treatment led to the extraction of lipids and are therefore not recommended.

Mechanical Behavior of Tough Hydrogels for Structural Applications

NASA Astrophysics Data System (ADS)

Illeperuma, Widusha Ruwangi Kaushalya

Hydrogels are widely used in many commercial products including Jell-O, contact lenses, and superabsorbent diapers. In recent decades, hydrogels have been under intense development for biomedical applications, such as scaffolds in tissue engineering, carriers for drug delivery, and valves in microfluidic systems. But the scope is severely limited as conventional hydrogels are weak and brittle and are not very stretchable. This thesis investigates the approaches that enhance the mechanical properties of hydrogels and their structural applications. We discov¬ered a class of exceptionally stretchable and tough hydrogels made from poly-mers that form networks via ionic and covalent crosslinks. Although such a hydrogel contains ~90% water, it can be stretched beyond 20 times its initial length, and has a fracture energy of ~9000 J/m2. The combination of large stretchability, remarkable toughness, and recoverability of stiffness and toughness, along with easy synthesis makes this material much superior over existing hydrogels. Extreme stretchability and blunted crack tips of these hydrogels question the validity of traditional fracture testing methods. We re-examine a widely used pure shear test method to measure the fracture energy. With the experimental and simulation results, we conclude that the pure shear test method can be used to measure fracture energy of extremely stretchable materials. Even though polyacrylamide-alginate hydrogels have an extremely high toughness, it has a relatively low stiffness and strength. We improved the stiffness and strength by embedding fibers. Most hydrogels are brittle, allowing the fibers to cut through the hydrogel when the composite is loaded. But tough hydrogel composites do not fail by the fibers cutting the hydrogel; instead, it undergoes large deforming by fibers sliding through the matrix. Hydrogels were not considered as materials for structural applications. But with enhanced mechanical properties, they have opened up novel applications. This thesis aims to investigate the broader applications, well beyond those investigated so far. We show fiber reinforced tough hydrogels can dissipate a significant amount of energy at a tunable level of stress, making them suitable for energy absorbing applications such as inner layer of helmets. We develop inexpensive fire-retarding materials using tough hydrogels that provide superior protection from burn injuries. We also study hydrogels as actuators that can be used in soft robotics. Hydrogels contain mostly water and they freeze when the temperature drops below 00C and lose its functions. We demonstrate a new class of hydrogels that do not freeze and hydrogels that partially freeze below water freezing temperature. Partially freezing hydrogels are ideal for cooling applications such as gel packs and non-freezing hydrogels are useful in all the structural applications at low temperatures. This thesis will enable the use of inexpensive hydrogels in a new class of non-traditional structural applications where the mechanical behavior of the hydrogel is of prime importance.

Investigation of features of plastic deformation and fracture of fine-crystalline V-4Ti-4Cr alloy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Grinyaev, Konstantin V., E-mail: kvgrinyaev@inbox.ru; Tyumentsev, Alexander N., E-mail: tyuments@phys.tsu.ru; Siberian Physical-Technical Institute, Tomsk, 634050

2014-11-14

With the use of transmission electron microscopy the investigation of defect substructure was carried out in the V-4Ti-4Cr-(C, N, O) alloy with disperse strengthening (by nanoparticles of oxy-carbo-nitride phase) after deformation by active tension at temperatures of 20 and 800 °C. It has been shown that an important feature of plastic deformation is deformation localization with crystal lattice reorientation.

The Processing and Mechanical Properties of High Temperature/ High Performance Composites. Stress Redistribution and Notch Properties. Book 2

DTIC Science & Technology

1994-03-01

unidirectional materials (Zok, Suo). In all cases, the mechanical measurements will be augmented by in-situ observations to identify changes in damage...be augmented by calculations of crack growth and fracture, incorporating the effects of thermal and elastic mismatch between the cladding and the...matrix cracking.1 5-2 1 All of these methods may be augmented by conventional optical and scanning electron microscopy. In the present study, a

Room Temperature Elastic Moduli and Vickers Hardness of Hot-Pressed LLZO Cubic Garnet

DTIC Science & Technology

2012-01-01

polishing compounds, Leco, St. Joseph, MI). X - ray diffraction and scanning electron microscopy (SEM) The microstructure of the hot-pressed specimens...was examined on uncoated fracture surfaces by SEM with an accelerating voltage of 1 and 3 kV. Phase purity was evaluated from X - ray diffraction data...the micro- structure appeared to be homogenous for the two hot- pressed LLZO specimens included in this study (Fig. 1). X - ray diffraction confirmed that

Multiscale Multiphysics Caprock Seal Analysis: A Case Study of the Farnsworth Unit, Texas, USA

NASA Astrophysics Data System (ADS)

Heath, J. E.; Dewers, T. A.; Mozley, P.

2015-12-01

Caprock sealing behavior depends on coupled processes that operate over a variety of length and time scales. Capillary sealing behavior depends on nanoscale pore throats and interfacial fluid properties. Larger-scale sedimentary architecture, fractures, and faults may govern properties of potential "seal-bypass" systems. We present the multiscale multiphysics investigation of sealing integrity of the caprock system that overlies the Morrow Sandstone reservoir, Farnsworth Unit, Texas. The Morrow Sandstone is the target injection unit for an on-going combined enhanced oil recovery-CO2 storage project by the Southwest Regional Partnership on Carbon Sequestration (SWP). Methods include small-to-large scale measurement techniques, including: focused ion beam-scanning electron microscopy; laser scanning confocal microscopy; electron and optical petrography; core examinations of sedimentary architecture and fractures; geomechanical testing; and a noble gas profile through sealing lithologies into the reservoir, as preserved from fresh core. The combined data set is used as part of a performance assessment methodology. The authors gratefully acknowledge the U.S. Department of Energy's (DOE) National Energy Technology Laboratory for sponsoring this project through the SWP under Award No. DE-FC26-05NT42591. Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

Adrenocortical Gap Junctions and Their Functions

PubMed Central

Bell, Cheryl L.; Murray, Sandra A.

2016-01-01

Adrenal cortical steroidogenesis and proliferation are thought to be modulated by gap junction-mediated direct cell–cell communication of regulatory molecules between cells. Such communication is regulated by the number of gap junction channels between contacting cells, the rate at which information flows between these channels, and the rate of channel turnover. Knowledge of the factors regulating gap junction-mediated communication and the turnover process are critical to an understanding of adrenal cortical cell functions, including development, hormonal response to adrenocorticotropin, and neoplastic dedifferentiation. Here, we review what is known about gap junctions in the adrenal gland, with particular attention to their role in adrenocortical cell steroidogenesis and proliferation. Information and insight gained from electrophysiological, molecular biological, and imaging (immunocytochemical, freeze fracture, transmission electron microscopic, and live cell) techniques will be provided. PMID:27445985

Recent developments in smart freezing technology applied to fresh foods.

PubMed

Xu, Ji-Cheng; Zhang, Min; Mujumdar, Arun S; Adhikari, Benu

2017-09-02

Due to the increased awareness of consumers in sensorial and nutritional quality of frozen foods, the freezing technology has to seek new and innovative technologies for better retaining the fresh like quality of foods. In this article, we review the recent developments in smart freezing technology applied to fresh foods. The application of these intelligent technologies and the associated underpinning concepts have greatly improved the quality of frozen foods and the freezing efficiency. These technologies are able to automatically collect the information in-line during freezing and help control the freezing process better. Smart freezing technology includes new and intelligent technologies and concepts applied to the pretreatment of the frozen product, freezing processes, cold chain logistics as well as warehouse management. These technologies enable real-time monitoring of quality during the freezing process and help improve product quality and freezing efficiency. We also provide a brief overview of several sensing technologies used to achieve automatic control of individual steps of freezing process. These sensing technologies include computer vision, electronic nose, electronic tongue, digital simulation, confocal laser, near infrared spectroscopy, nuclear magnetic resonance technology and ultrasound. Understanding of the mechanism of these new technologies will be helpful for applying them to improve the quality of frozen foods.

Radiographic and ultrasonic characterization of sintered silicon carbide

NASA Technical Reports Server (NTRS)

Baaklini, G. Y.; Abel, P. B.

1988-01-01

The capabilities were investigated of projection microfocus X-radiography, ultrasonic velocity and attenuation, and reflection scanning acoustic microscopy for characterizing silicon carbide specimens. Silicon carbide batches covered a range of densities and different microstructural characteristics. Room temperature, four point flexural strength tests were conducted. Fractography was used to identify types, sizes, and locations of fracture origins. Fracture toughness values were calculated from fracture strength and flaw characterization data. Detection capabilities of radiography and acoustic microscopy for fracture-causing flaws were evaluated. Applicability of ultrasonics for verifying material strength and toughness was examined.

Flaw imaging and ultrasonic techniques for characterizing sintered silicon carbide

NASA Technical Reports Server (NTRS)

Baaklini, George Y.; Abel, Phillip B.

1987-01-01

The capabilities were investigated of projection microfocus x-radiography, ultrasonic velocity and attenuation, and reflection scanning acoustic microscopy for characterizing silicon carbide specimens. Silicon carbide batches covered a range of densities and different microstructural characteristics. Room temperature, four point flexural strength tests were conducted. Fractography was used to identify types, sizes, and locations of fracture origins. Fracture toughness values were calculated from fracture strength and flaw characterization data. Detection capabilities of radiography and acoustic microscopy for fracture-causing flaws were evaluated. Applicability of ultrasonics for verifying material strength and toughness was examined.

Self-healing of cracks formed in Silicon-Aluminum anodes electrochemically cycled at high lithiation rates

NASA Astrophysics Data System (ADS)

Bhattacharya, Sandeep; Alpas, Ahmet T.

2016-10-01

Lithiation-induced volume changes in Si result in fracture and fragmentation of Si anodes in Li-ion batteries. This paper reports the self-healing behaviour of cracks observed in micron-sized Si particles dispersed in a ductile Al matrix of a Si-Al electrode electrochemically cycled vs. Li/Li+ using a high lithiation rate of 15.6 C. Cross-sectional high-resolution transmission electron microscopy and Raman spectroscopy revealed that an amorphous layer with a depth up to ∼100 nm was formed at the surface of Si particles. In-situ optical microscopy performed during electrochemical experiments revealed development of cracks in Si particles as the voltage decreased to 0.02 V during lithiation. Self-healing of cracks in Si particles occurred in two steps: i) arresting of the crack growth at the Si/Al interface as the surrounding Al matrix had a higher fracture toughness and thus acted as a barrier to crack propagation, and ii) closure of cracks due to compressive stresses applied to the crack faces by the amorphous zones formed on each side of the crack paths.

Fracture Surface Morphology and Impact Strength of Cellulose/PLA Composites.

PubMed

Gao, Honghong; Qiang, Tao

2017-06-07

Polylactide (PLA)-based composite materials reinforced with ball-milled celluloses were manufactured by extrusion blending followed by injection molding. Their surface morphology from impact fracture were imaged with scanning electron microscopy (SEM) and investigated by calculating their fractal dimensions. Then, linear regression was used to explore the relationship between fractal dimension and impact strength of the resultant cellulose/PLA composite materials. The results show that filling the ball-milled celluloses into PLA can improve the impact toughness of PLA by a minimum of 38%. It was demonstrated that the fracture pattern of the cellulose/PLA composite materials is different from that of pristine PLA. For the resultant composite materials, the fractal dimension of the impact fractured surfaces increased with increasing filling content and decreasing particle size of the ball-milled cellulose particles. There were highly positive correlations between fractal dimension of the fractured surfaces and impact strength of the cellulose/PLA composites. However, the linearity between fractal dimension and impact strength were different for the different methods, due to their different R-squared values. The approach presented in this work will help to understand the structure-property relationships of composite materials from a new perspective.

Clinical Evaluation of an Unsintered Hydroxyapatite/Poly-L-Lactide Osteoconductive Composite Device for the Internal Fixation of Maxillofacial Fractures

PubMed Central

Sukegawa, Shintaro; Kanno, Takahiro; Katase, Naoki; Shibata, Akane; Takahashi, Yuka; Furuki, Yoshihiko

2016-01-01

Introduction: OSTEOTRANS MX (Takiron Co, Ltd, Osaka, Japan) is a resorbable osteosynthetic material composed of an unsintered hydroxyapatite/poly-l-lactide composite, and its osteoconductive capacity has been documented. The authors here report their clinical experience using OSTEOTRANS MX. Methods: The authors treated 35 patients (19 men, 16 women; age, 14–88 years; mean ± standard deviation, 38.4 ± 19.9 years) with maxillofacial fractures. The authors used standard surgery to stabilize fractures in all patients, fitting resorbable plates (thickness, 1.0 or 1.4 mm) and screws (diameter, 2 mm) according to Arbeitsgemeinschaft für Osteosynthesefragen/Association (AO) for the Study of Internal Fixation guidelines. Results: All patients eventually achieved satisfactory healing with favorable restoration of form and function without foreign body reaction. Complications occurred in 3 patients—plate exposure in 2 and discomfort in 1. However, fracture sites healed in all patients. Scanning electron microscopy revealed that the devices bonded directly to the bone without interposition of nonmineralized tissue. Conclusion: OSTEOTRANS MX is a useful material with few complications. Its osteoconductive bioactivity is advantageous for the early functional improvement of maxillofacial fractures. PMID:27428913

Fracture Surface Morphology and Impact Strength of Cellulose/PLA Composites

PubMed Central

Gao, Honghong; Qiang, Tao

2017-01-01

Polylactide (PLA)-based composite materials reinforced with ball-milled celluloses were manufactured by extrusion blending followed by injection molding. Their surface morphology from impact fracture were imaged with scanning electron microscopy (SEM) and investigated by calculating their fractal dimensions. Then, linear regression was used to explore the relationship between fractal dimension and impact strength of the resultant cellulose/PLA composite materials. The results show that filling the ball-milled celluloses into PLA can improve the impact toughness of PLA by a minimum of 38%. It was demonstrated that the fracture pattern of the cellulose/PLA composite materials is different from that of pristine PLA. For the resultant composite materials, the fractal dimension of the impact fractured surfaces increased with increasing filling content and decreasing particle size of the ball-milled cellulose particles. There were highly positive correlations between fractal dimension of the fractured surfaces and impact strength of the cellulose/PLA composites. However, the linearity between fractal dimension and impact strength were different for the different methods, due to their different R-squared values. The approach presented in this work will help to understand the structure–property relationships of composite materials from a new perspective. PMID:28772983

Influence of surface morphology on the immersion mode ice nucleation efficiency of hematite particles

NASA Astrophysics Data System (ADS)

Hiranuma, N.; Hoffmann, N.; Kiselev, A.; Dreyer, A.; Zhang, K.; Kulkarni, G.; Koop, T.; Möhler, O.

2014-03-01

In this paper, the effect of the morphological modification of aerosol particles with respect to heterogeneous ice nucleation is comprehensively investigated for laboratory-generated hematite particles as a model substrate for atmospheric dust particles. The surface-area-scaled ice nucleation efficiencies of monodisperse cubic hematite particles and milled hematite particles were measured with a series of expansion cooling experiments using the Aerosol Interaction and Dynamics in the Atmosphere (AIDA) cloud simulation chamber. Complementary offline characterization of physico-chemical properties of both hematite subsets were also carried out with scanning electron microscopy (SEM), energy dispersive X-ray (EDX) spectroscopy, dynamic light scattering (DLS), and an electro-kinetic particle charge detector to further constrain droplet-freezing measurements of hematite particles. Additionally, an empirical parameterization derived from our laboratory measurements was implemented in the single-column version of the Community Atmospheric Model version 5 (CAM5) to investigate the model sensitivity in simulated ice crystal number concentration on different ice nucleation efficiencies. From an experimental perspective, our results show that the immersion mode ice nucleation efficiency of milled hematite particles is almost an order of magnitude higher at -35.2 °C < T < -33.5 °C than that of the cubic hematite particles, indicating a substantial effect of morphological irregularities on immersion mode freezing. Our modeling results similarly show that the increased droplet-freezing rates of milled hematite particles lead to about one order magnitude higher ice crystal number in the upper troposphere than cubic hematite particles. Overall, our results suggest that the surface irregularities and associated active sites lead to greater ice activation through droplet freezing.

Freeze-Thaw Cycle Test on Basalt, Diorite and Tuff Specimens with the Simulated Ground Temperature of Antarctica

NASA Astrophysics Data System (ADS)

Park, J.; Hyun, C.; Cho, H.; Park, H.

2010-12-01

Physical weathering caused by freeze-thaw action in cold regions was simulated with artificial weathering simulator in laboratory. Physical weathering of rock in cold regions usually depends on the temperature, rock type and moisture content. Then these three variables were considered in this study. The laboratory freeze-thaw tests were conducted on the three types of rocks, e.g. diorite, basalt and tuff, which are the major rock types around Sejong Station, King George Island, Antarctica. Nine core samples composed of three samples from each rock type were prepared in NX core, and 50 cycles of freeze-thaw test was carried out under dried and saturated water conditions. In this study, the physical weathering of rocks was investigated after each 10 cycles by measuring P-wave velocity, bulk density, effective porosity, Schmidt hardness and uniaxial compression strength(UCS). The experimental result of the diorite and the tuff specimens showed that P-wave velocity, bulk density, effective porosity, Schmidt hardness and UCS were gradually decreased as weathering progresses, but the result of the basalt specimens did not show typical trends due to the characteristics of irregular pore distribution and various pore sizes. Scanning electron microscopy(SEM) photographs of diorite, basalt and tuff specimens weathered in dried and saturated conditions were also acquired to investigate the role of water during physical weathering processes. The number and size of microcracks were increased as weathering progresses. This work was supported by the National Research Foundation of Korea(NRF) Grant(NRF-2010-0027753).

Heterogeneous ice nucleation of α-pinene SOA particles before and after ice cloud processing

NASA Astrophysics Data System (ADS)

Wagner, Robert; Höhler, Kristina; Huang, Wei; Kiselev, Alexei; Möhler, Ottmar; Mohr, Claudia; Pajunoja, Aki; Saathoff, Harald; Schiebel, Thea; Shen, Xiaoli; Virtanen, Annele

2017-05-01

The ice nucleation ability of α-pinene secondary organic aerosol (SOA) particles was investigated at temperatures between 253 and 205 K in the Aerosol Interaction and Dynamics in the Atmosphere cloud simulation chamber. Pristine SOA particles were nucleated and grown from pure gas precursors and then subjected to repeated expansion cooling cycles to compare their intrinsic ice nucleation ability during the first nucleation event with that observed after ice cloud processing. The unprocessed α-pinene SOA particles were found to be inefficient ice-nucleating particles at cirrus temperatures, with nucleation onsets (for an activated fraction of 0.1%) as high as for the homogeneous freezing of aqueous solution droplets. Ice cloud processing at temperatures below 235 K only marginally improved the particles' ice nucleation ability and did not significantly alter their morphology. In contrast, the particles' morphology and ice nucleation ability was substantially modified upon ice cloud processing in a simulated convective cloud system, where the α-pinene SOA particles were first activated to supercooled cloud droplets and then froze homogeneously at about 235 K. As evidenced by electron microscopy, the α-pinene SOA particles adopted a highly porous morphology during such a freeze-drying cycle. When probing the freeze-dried particles in succeeding expansion cooling runs in the mixed-phase cloud regime up to 253 K, the increase in relative humidity led to a collapse of the porous structure. Heterogeneous ice formation was observed after the droplet activation of the collapsed, freeze-dried SOA particles, presumably caused by ice remnants in the highly viscous material or the larger surface area of the particles.

Preparation of finasteride capsules-loaded drug nanoparticles: formulation, optimization, in vitro, and pharmacokinetic evaluation

PubMed Central

Ahmed, Tarek A

2016-01-01

In this study, optimized freeze-dried finasteride nanoparticles (NPs) were prepared from drug nanosuspension formulation that was developed using the bottom–up technique. The effects of four formulation and processing variables that affect the particle size and solubility enhancement of the NPs were explored using the response surface optimization design. The optimized formulation was morphologically characterized using transmission electron microscopy (TEM). Physicochemical interaction among the studied components was investigated. Crystalline change was investigated using X-ray powder diffraction (XRPD). Crystal growth of the freeze-dried NPs was compared to the corresponding aqueous drug nanosuspension. Freeze-dried NPs formulation was subsequently loaded into hard gelatin capsules that were examined for in vitro dissolution and pharmacokinetic behavior. Results revealed that in most of the studied variables, some of the quadratic and interaction effects had a significant effect on the studied responses. TEM image illustrated homogeneity and shape of the prepared NPs. No interaction among components was noticed. XRPD confirmed crystalline state change in the optimized NPs. An enhancement in the dissolution rate of more than 2.5 times from capsules filled with optimum drug NPs, when compared to capsules filled with pure drug, was obtained. Crystal growth, due to Ostwald ripening phenomenon and positive Gibbs free energy, was reduced following lyophilization of the nanosuspension formulation. Pharmacokinetic parameters from drug NPs were superior to that of pure drug and drug microparticles. In conclusion, freeze-dried NPs based on drug nanosuspension formulation is a successful technique in enhancing stability, solubility, and in vitro dissolution of poorly water-soluble drugs with possible impact on the drug bioavailability. PMID:26893559

Influence of rotational speed on the cyclic fatigue of rotary nickel-titanium endodontic instruments.

PubMed

Lopes, Hélio P; Ferreira, Alessandra A P; Elias, Carlos N; Moreira, Edson J L; de Oliveira, Júlio C Machado; Siqueira, José F

2009-07-01

During the preparation of curved canals, rotary nickel-titanium (NiTi) instruments are subjected to cyclic fatigue, which can lead to instrument fracture. Although several factors may influence the cyclic fatigue resistance of instruments, the role of the rotational speed remains uncertain. This study was intended to evaluate the effects of rotational speed on the number of cycles to fracture of rotary NiTi instruments. ProTaper Universal instruments F3 and F4 (Maillefer SA, Ballaigues, Switzerland) were used in an artificial curved canal under rotational speeds of 300 rpm or 600 rpm. The artificial canal was made of stainless steel, with an inner diameter of 1.5 mm, total length of 20 mm, and arc at the end with a curvature radius of 6 mm. The arc length was 9.4 mm and 10.6 mm on the straight part. The number of cycles required to fracture was recorded. Fractured surfaces and the helical shafts of the fractured instruments were analyzed by scanning electron microscopy. The results showed approximately a 30% reduction in the observed number of cycles to fracture as rotational speed was increased from 300 to 600 RPM (p < 0.05). The morphology of the fractured surface was always of ductile type, and no plastic deformation was observed on the helical shaft of fractured instruments. The present findings for both F3 and F4 ProTaper instruments revealed that the increase in rotational speed significantly reduced the number of cycles to fracture.

Laser Engineered Net Shaping of Nickel-Based Superalloy Inconel 718 Powders onto AISI 4140 Alloy Steel Substrates: Interface Bond and Fracture Failure Mechanism

PubMed Central

Kim, Hoyeol; Cong, Weilong; Zhang, Hong-Chao; Liu, Zhichao

2017-01-01

As a prospective candidate material for surface coating and repair applications, nickel-based superalloy Inconel 718 (IN718) was deposited on American Iron and Steel Institute (AISI) 4140 alloy steel substrate by laser engineered net shaping (LENS) to investigate the compatibility between two dissimilar materials with a focus on interface bonding and fracture behavior of the hybrid specimens. The results show that the interface between the two dissimilar materials exhibits good metallurgical bonding. Through the tensile test, all the fractures occurred in the as-deposited IN718 section rather than the interface or the substrate, implying that the as-deposited interlayer bond strength is weaker than the interfacial bond strength. From the fractography using scanning electron microscopy (SEM) and energy disperse X-ray spectrometry (EDS), three major factors affecting the tensile fracture failure of the as-deposited part are (i) metallurgical defects such as incompletely melted powder particles, lack-of-fusion porosity, and micropores; (ii) elemental segregation and Laves phase, and (iii) oxide formation. The fracture failure mechanism is a combination of all these factors which are detrimental to the mechanical properties and structural integrity by causing premature fracture failure of the as-deposited IN718. PMID:28772702

Influence of curvature location along an artificial canal on cyclic fatigue of a rotary nickel-titanium endodontic instrument.

PubMed

Lopes, Hélio Pereira; Chiesa, Wanderson Miguel Maia; Correia, Norbi Rodrigues; de Souza Navegante, Núbia Carvalho; Elias, Carlos Nelson; Moreira, Edson Jorge Lima; Chiesa, Bianca Esther Cabral

2011-06-01

This study evaluated the effects of curvature location along an artificial canal on cyclic fatigue (CF) of an Mtwo rotary instrument, verifying the number of cycles to fatigue fracture (NCF) and morphologic characteristics of the fractured instruments. CF testing of instruments was performed in artificial canals with curvature radii of 10 mm and arc lengths of 11 mm. Mtwo rotary instruments size 40, 0.04 taper were used in 2 groups (n = 10): group A, curvature positioned on middle part; group B, apical curvature. All instruments were rotated until fracture. The number of cycles to failure was registered. Data were analyzed by independent sample t test. Fractured surfaces and the helical shafts of the instruments were analyzed by scanning electron microscopy. NCF for groups A and B had significant statistic differences (P < .05). The highest values were found in the group where the curvature was positioned on the apical part of the canal. No plastic deformation was observed on the helical shafts. The number of cycles to fracture of the Mtwo instruments increased when the arc was changed from the middle to the apical part of the canal. The morphologic characteristics of the fractured surfaces were of the ductile type. Copyright © 2011 Mosby, Inc. All rights reserved.

Quantifying water flow and retention in an unsaturated fracture-facial domain

USGS Publications Warehouse

Nimmo, John R.; Malek-Mohammadi, Siamak

2015-01-01

Hydrologically significant flow and storage of water occur in macropores and fractures that are only partially filled. To accommodate such processes in flow models, we propose a three-domain framework. Two of the domains correspond to water flow and water storage in a fracture-facial region, in addition to the third domain of matrix water. The fracture-facial region, typically within a fraction of a millimeter of the fracture wall, includes a flowing phase whose fullness is determined by the availability and flux of preferentially flowing water, and a static storage portion whose fullness is determined by the local matric potential. The flow domain can be modeled with the source-responsive preferential flow model, and the roughness-storage domain can be modeled with capillary relations applied on the fracture-facial area. The matrix domain is treated using traditional unsaturated flow theory. We tested the model with application to the hydrology of the Chalk formation in southern England, coherently linking hydrologic information including recharge estimates, streamflow, water table fluctuation, imaging by electron microscopy, and surface roughness. The quantitative consistency of the three-domain matrix-microcavity-film model with this body of diverse data supports the hypothesized distinctions and active mechanisms of the three domains and establishes the usefulness of this framework.

Laser Engineered Net Shaping of Nickel-Based Superalloy Inconel 718 Powders onto AISI 4140 Alloy Steel Substrates: Interface Bond and Fracture Failure Mechanism.

PubMed

Kim, Hoyeol; Cong, Weilong; Zhang, Hong-Chao; Liu, Zhichao

2017-03-25

As a prospective candidate material for surface coating and repair applications, nickel-based superalloy Inconel 718 (IN718) was deposited on American Iron and Steel Institute (AISI) 4140 alloy steel substrate by laser engineered net shaping (LENS) to investigate the compatibility between two dissimilar materials with a focus on interface bonding and fracture behavior of the hybrid specimens. The results show that the interface between the two dissimilar materials exhibits good metallurgical bonding. Through the tensile test, all the fractures occurred in the as-deposited IN718 section rather than the interface or the substrate, implying that the as-deposited interlayer bond strength is weaker than the interfacial bond strength. From the fractography using scanning electron microscopy (SEM) and energy disperse X-ray spectrometry (EDS), three major factors affecting the tensile fracture failure of the as-deposited part are (i) metallurgical defects such as incompletely melted powder particles, lack-of-fusion porosity, and micropores; (ii) elemental segregation and Laves phase, and (iii) oxide formation. The fracture failure mechanism is a combination of all these factors which are detrimental to the mechanical properties and structural integrity by causing premature fracture failure of the as-deposited IN718.

Synthesis and Properties of a Clean and Sustainable Deicing Additive for Asphalt Mixture

PubMed Central

Peng, Chao; Yu, Jianying; Zhao, Zhijie; Dai, Jing; Fu, Jingyi; Zhao, Meiling; Wang, Wei

2015-01-01

A clean and sustainable deicing additive was prepared via the adsorption of acetate anions (Ac-) by magnesium (Mg) and aluminum (Al) calcined layered double hydroxide (Mg/Al-CLDH). Fourier transform infrared spectroscopy spectrums proved that Ac- had intercalated into LDH structure. X-ray diffraction patterns, scanning electron microscopy and transmission electron microscopy images showed that the intercalation spacing and platelet thickness of Mg and Al layered double hydroxide containing Ac- anions (Mg/Al-Ac- LDH) had been enlarged due to substitution of divalent CO3 2- anions by a larger quantity of monovalent Ac– anions. Differential scanning calorimetry tests testified that the insoluble Mg2/Al-Ac- LDH evidently decreased the freeze point (FP) of water to -10.68°C. X-ray photoelectron spectroscopy analyses confirmed that the Ac- were strongly confined by the metal layers of LDHs. FP test of asphalt mixtures confirmed that Mg/Al-Ac- LDHs reduced FP to -5.5°C. Immersion test results indicated that Mg/Al-Ac- LDH had a good deicing durability and Ac- did not released from asphalt mixture. Snow melting observation was conducted further testified that Mg/Al-Ac- LDH melted snow or ice sustainably. PMID:25625279

Synthesis and properties of a clean and sustainable deicing additive for asphalt mixture.

PubMed

Peng, Chao; Yu, Jianying; Zhao, Zhijie; Dai, Jing; Fu, Jingyi; Zhao, Meiling; Wang, Wei

2015-01-01

A clean and sustainable deicing additive was prepared via the adsorption of acetate anions (Ac-) by magnesium (Mg) and aluminum (Al) calcined layered double hydroxide (Mg/Al-CLDH). Fourier transform infrared spectroscopy spectrums proved that Ac- had intercalated into LDH structure. X-ray diffraction patterns, scanning electron microscopy and transmission electron microscopy images showed that the intercalation spacing and platelet thickness of Mg and Al layered double hydroxide containing Ac- anions (Mg/Al-Ac- LDH) had been enlarged due to substitution of divalent CO32- anions by a larger quantity of monovalent Ac- anions. Differential scanning calorimetry tests testified that the insoluble Mg2/Al-Ac- LDH evidently decreased the freeze point (FP) of water to -10.68°C. X-ray photoelectron spectroscopy analyses confirmed that the Ac- were strongly confined by the metal layers of LDHs. FP test of asphalt mixtures confirmed that Mg/Al-Ac- LDHs reduced FP to -5.5°C. Immersion test results indicated that Mg/Al-Ac- LDH had a good deicing durability and Ac- did not released from asphalt mixture. Snow melting observation was conducted further testified that Mg/Al-Ac- LDH melted snow or ice sustainably.

Petrological features of selected components of the Cergowa sandstones (Outer Carpathians) recorded by scanning electron microscopy - preliminary study

NASA Astrophysics Data System (ADS)

Pszonka, Joanna

2017-11-01

The scanning electron microscope analysis of the Cergowa sandstones brings new data on their petrological features and chemical composition. Previous work in standard petrographic examination, e.g. polarising (PL) or cathodoluminescence (CL) microscopy, displayed limited information on grain surface topography and only assumptions to their geochemistry. Both identification and characterisation of minerals are fundamental in the progress of mining and minerals processing systems. Detrital grains of the Cergowa sandstones are bound by calcite and dolomitic cement and commonly corroded by diagenetic fluids, however, in varying degrees, which is illustrated here by feldspar, quartz and dolomite minerals. Dissolution processes of marginal parts of these mineral grains resulted in corrosion, which increased the contact surface between the grains and the cement. The difference in resistance to these processes was observed not only among distinct groups of minerals, but also within the group of feldspars: between K-feldspars and minerals of plagioclase. That combination resulted in exceptionally strong cementation of the Cergowa sandstones, which is expressed by their high hardness and resistance to abrasion, freezing, and thawing. Inherent parameters of sandstones are characterised by their petrographical properties.

Pickering emulsions stabilized by a metal-organic framework (MOF) and graphene oxide (GO) for producing MOF/GO composites.

PubMed

Zhang, Fanyu; Liu, Lifei; Tan, Xiuniang; Sang, Xinxin; Zhang, Jianling; Liu, Chengcheng; Zhang, Bingxing; Han, Buxing; Yang, Guanying

2017-10-18

Herein we demonstrate the formation of a novel kind of Pickering emulsion that is stabilized by a Zr-based metal-organic framework (Zr-MOF) and graphene oxide (GO). It was found that the Zr-BDC-NO 2 and GO solids assembling at the oil/water interface can effectively stabilize the oil droplets that are dispersed in the water phase. Such a Pickering emulsion offers a facile route for fabricating Zr-MOF/GO composite materials. After removing water and oil by freeze drying from Pickering emulsions, the Zr-MOF/GO composites were obtained and their morphologies, structures and interaction properties were characterized by scanning electron microscopy, transmission electron microscopy, X-ray diffraction and Fourier transform infrared spectrometry, respectively. The influences of the concentration of GO and Zr-MOF on the emulsion microstructures and the properties of the MOF/GO composites were studied. Based on experimental results, the mechanisms for the emulsion formation by Zr-MOF and GO and the as-synthesized superstructures of the Zr-MOF/GO composite were proposed. It is expected that this facile and tunable route can be applied to the synthesis of different kinds of MOF-based or GO-based composite materials.

Effects of surface treatments and bonding types on the interfacial behavior of fiber metal laminate based on magnesium alloy

NASA Astrophysics Data System (ADS)

Zhang, Xi; Ma, Quanyang; Dai, Yu; Hu, Faping; Liu, Gang; Xu, Zouyuan; Wei, Guobing; Xu, Tiancai; Zeng, Qingwen; Xie, Weidong

2018-01-01

Fiber metal laminates based on magnesium alloys (MgFML) with different surface treatments and different bonding types were tested and analyzed. By using dynamic contact angle measurement and scanning electron microscopy (SEM), it was found that phosphating treatment can significantly improve the surface energy and wettability of magnesium alloy, and the surface energy of phosphated magnesium alloy was approximately 50% higher than that of abraded-only magnesium alloy. The single cantilever beam (SCB) test showed that the interfacial fracture energies of directly bonded MgFMLs based on abraded-only magnesium and abraded + phosphated magnesium were 650 J/m2 and 1030 J/m2, respectively, whereas the interfacial fracture energies of indirectly bonded MgFMLs were 1650 J/m2 and 2260 J/m2, respectively. Phosphating treatment and modified polypropylene interleaf were observed to improve the tensile strength and interfacial fracture toughness of MgFML. In addition, the rougher surface was more conducive to enhance the bonding strength and interfacial fracture toughness of MgFML.

The fracture behaviour of dental enamel.

PubMed

Bechtle, Sabine; Habelitz, Stefan; Klocke, Arndt; Fett, Theo; Schneider, Gerold A

2010-01-01

Enamel is the hardest tissue in the human body covering the crowns of teeth. Whereas the underlying dental material dentin is very well characterized in terms of mechanical and fracture properties, available data for enamel are quite limited and are apart from the most recent investigation mainly based on indentation studies. Within the current study, stable crack-growth experiments in bovine enamel have been performed, to measure fracture resistance curves for enamel. Single edge notched bending specimens (SENB) prepared out of bovine incisors were tested in 3-point bending and subsequently analysed using optical and environmental scanning electron microscopy. Cracks propagated primarily within the protein-rich rod sheaths and crack propagation occurred under an inclined angle to initial notch direction not only due to enamel rod and hydroxyapatite crystallite orientation but potentially also due to protein shearing. Determined mode I fracture resistance curves ranged from 0.8-1.5 MPa*m(1/2) at the beginning of crack propagation up to 4.4 MPa*m(1/2) at 500 microm crack extension; corresponding mode II values ranged from 0.3 to 1.5 MPa*m(1/2).

Ion microprobe, electron microprobe and cathodoluminescence data for Allende inclusions with emphasis on plagioclase chemistry

NASA Technical Reports Server (NTRS)

Hutcheon, I. D.; Steele, I. M.; Smith, J. V.; Clayton, R. N.

1978-01-01

Three Type B inclusions from the Allende meteorite have been analyzed. A grain-to-grain characterization of mineral chemistry and isotopic content was made possible by the use of a range of techniques, including luminescence and scanning electron microscopy and electron and ion microprobe analysis. Cathodoluminescence was used in fine-grained, optically opaque regions to distinguish between sub-micrometer phases, such as garnet and Si-rich material, subsequently identified by electron probe and scanning electron microscope analyses. Four types of luminescence patterns, due to twinning, primary sector zoning, alteration of boundaries and fractures, and shock effects, were identified in Allende plagioclase. Luminescence color exhibited a strong correlation with Mg content and provided a guide for an electron probe quantitative map of Mg and Na distributions. Ion microprobe studies of individual grains revealed large excesses of Mg-26.

The Effect of Freezing on the Dynamics of Dike Propagation

NASA Astrophysics Data System (ADS)

Tait, S.; Taisne, B.

2007-12-01

When magma-filled cracks propagate close to the Earth's surface, host rock temperature is well below the magma solidus. Solidification and substantial increase in magma viscosity can occur, are most pronounced near the propagating tip and can slow or arrest the progress of the dike. Quantitative analysis is required to predict whether a given dike will reach the surface to erupt and the duration of the precursor sequence. This challenging physical problem mixes elasticity, fracture mechanics, heat transfer and fluid flow with strong rheologic gradients due to cooling. We describe the propagation behaviour of such a hydraulic fracture using a laboratory experimental system of a crack fed by a constant flux of paraffin wax from a source reservoir propagating through gelatin below the solidus of the wax. The most novel behaviour is an intermittent regime in which cracks periodically stop advancing due to solidification, then swell at constant length while enhancing the elastic deformation in the surrounding solid before propagation resumes. We present a physical model of this system, based on different balances between driving and resistive forces: the former are elastic stress and liquid buoyancy, the latter are fracture resistance at the tip and viscous resistance. The fracture is represented as a head, behind the propagating tip, connected to the source by a narrow tail. Freezing of liquid close to the tip is assumed to enhance fracture resistance according to a cooling law, and propagation is assumed to occur only when the stress exerted by the liquid is enough to overcome fracture resistance. Our theoretical model reproduces intermittent propagation with precise behaviour depending on the controlling stress balances, and provides a tool to analyse natural systems.

Sodium selective ion channel formation in living cell membranes by polyamidoamine dendrimer.

PubMed

Nyitrai, Gabriella; Keszthelyi, Tamás; Bóta, Attila; Simon, Agnes; Tőke, Orsolya; Horváth, Gergő; Pál, Ildikó; Kardos, Julianna; Héja, László

2013-08-01

Polyamidoamine (PAMAM) dendrimers are highly charged hyperbranched protein-like polymers that are known to interact with cell membranes. In order to disclose the mechanisms of dendrimer-membrane interaction, we monitored the effect of PAMAM generation five (G5) dendrimer on the membrane permeability of living neuronal cells followed by exploring the underlying structural changes with infrared-visible sum frequency vibrational spectroscopy (SVFS), small angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). G5 dendrimers were demonstrated to irreversibly increase the membrane permeability of neurons that could be blocked in low-[Na(+)], but not in low-[Ca(2+)] media suggesting the formation of specific Na(+) permeable channels. SFVS measurements on silica supported DPPG-DPPC bilayers suggested G5-specific trans-polarization of the membrane. SAXS data and freeze-fracture TEM imaging of self-organized DPPC vesicle systems demonstrated disruption of DPPC vesicle layers by G5 through polar interactions between G5 terminal amino groups and the anionic head groups of DPPC. We propose a nanoscale mechanism by which G5 incorporates into the membrane through multiple polar interactions that disrupt proximate membrane bilayer and shape a unique hydrophilic Na(+) ion permeable channel around the dendrimer. In addition, we tested whether these artificial Na(+) channels can be exploited as antibiotic tools. We showed that G5 quickly arrest the growth of resistant bacterial strains below 10μg/ml concentration, while they show no detrimental effect on red blood cell viability, offering the chance for the development of new generation anti-resistant antibiotics. Copyright © 2013 Elsevier B.V. All rights reserved.

In tight junctions, claudins regulate the interactions between occludin, tricellulin and marvelD3, which, inversely, modulate claudin oligomerization.

PubMed

Cording, Jimmi; Berg, Johanna; Käding, Nadja; Bellmann, Christian; Tscheik, Christian; Westphal, Julie K; Milatz, Susanne; Günzel, Dorothee; Wolburg, Hartwig; Piontek, Jörg; Huber, Otmar; Blasig, Ingolf Ernst

2013-01-15

Tight junctions seal the paracellular cleft of epithelia and endothelia, form vital barriers between tissue compartments and consist of tight-junction-associated marvel proteins (TAMPs) and claudins. The function of TAMPs and the interaction with claudins are not understood. We therefore investigated the binding between the TAMPs occludin, tricellulin, and marvelD3 and their interaction with claudins in living tight-junction-free human embryonic kidney-293 cells. In contrast to claudins and occludin, tricellulin and marvelD3 showed no enrichment at cell-cell contacts indicating lack of homophilic trans-interaction between two opposing cell membranes. However, occludin, marvelD3 and tricellulin exhibited homophilic cis-interactions, along one plasma membrane, as measured by fluorescence resonance energy transfer. MarvelD3 also cis-interacted with occludin and tricellulin heterophilically. Classic claudins, such as claudin-1 to -5 may show cis-oligomerization with TAMPs, whereas the non-classic claudin-11 did not. Claudin-1 and -5 improved enrichment of occludin and tricellulin at cell-cell contacts. The low mobile claudin-1 reduced the membrane mobility of the highly mobile occludin and tricellulin, as studied by fluorescence recovery after photobleaching. Co-transfection of claudin-1 with TAMPs led to changes of the tight junction strand network of this claudin to a more physiological morphology, depicted by freeze-fracture electron microscopy. The results demonstrate multilateral interactions between the tight junction proteins, in which claudins determine the function of TAMPs and vice versa, and provide deeper insights into the tight junction assembly.

Mode of action of claudin peptidomimetics in the transient opening of cellular tight junction barriers.

PubMed

Staat, Christian; Coisne, Caroline; Dabrowski, Sebastian; Stamatovic, Svetlana M; Andjelkovic, Anuska V; Wolburg, Hartwig; Engelhardt, Britta; Blasig, Ingolf E

2015-06-01

In epithelial/endothelial barriers, claudins form tight junctions, seal the paracellular cleft, and limit the uptake of solutes and drugs. The peptidomimetic C1C2 from the C-terminal half of claudin-1's first extracellular loop increases drug delivery through epithelial claudin-1 barriers. However, its molecular and structural mode of action remains unknown. In the present study, >100 μM C1C2 caused paracellular opening of various barriers with different claudin compositions, ranging from epithelial to endothelial cells, preferentially modulating claudin-1 and claudin-5. After 6 h incubation, C1C2 reversibly increased the permeability to molecules of different sizes; this was accompanied by redistribution of claudins and occludin from junctions to cytosol. Internalization of C1C2 in epithelial cells depended on claudin-1 expression and clathrin pathway, whereby most C1C2 was retained in recyclosomes >2 h. In freeze-fracture electron microscopy, C1C2 changed claudin-1 tight junction strands to a more parallel arrangement and claudin-5 strands from E-face to P-face association - drastic and novel effects. In conclusion, C1C2 is largely recycled in the presence of a claudin, which explains the delayed onset of barrier and junction loss, the high peptide concentration required and the long-lasting effect. Epithelial/endothelial barriers are specifically modulated via claudin-1/claudin-5, which can be targeted to improve drug delivery. Copyright © 2015 Elsevier Ltd. All rights reserved.

Molecular Modeling and Imaging of Initial Stages of Cellulose Fibril Assembly: Evidence for a Disordered Intermediate Stage

PubMed Central

Haigler, Candace H.; Grimson, Mark J.; Gervais, Julien; Le Moigne, Nicolas; Höfte, Herman; Monasse, Bernard; Navard, Patrick

2014-01-01

The remarkable mechanical strength of cellulose reflects the arrangement of multiple β-1,4-linked glucan chains in a para-crystalline fibril. During plant cellulose biosynthesis, a multimeric cellulose synthesis complex (CSC) moves within the plane of the plasma membrane as many glucan chains are synthesized from the same end and in close proximity. Many questions remain about the mechanism of cellulose fibril assembly, for example must multiple catalytic subunits within one CSC polymerize cellulose at the same rate? How does the cellulose fibril bend to align horizontally with the cell wall? Here we used mathematical modeling to investigate the interactions between glucan chains immediately after extrusion on the plasma membrane surface. Molecular dynamics simulations on groups of six glucans, each originating from a position approximating its extrusion site, revealed initial formation of an uncrystallized aggregate of chains from which a protofibril arose spontaneously through a ratchet mechanism involving hydrogen bonds and van der Waals interactions between glucose monomers. Consistent with the predictions from the model, freeze-fracture transmission electron microscopy using improved methods revealed a hemispherical accumulation of material at points of origination of apparent cellulose fibrils on the external surface of the plasma membrane where rosette-type CSCs were also observed. Together the data support the possibility that a zone of uncrystallized chains on the plasma membrane surface buffers the predicted variable rates of cellulose polymerization from multiple catalytic subunits within the CSC and acts as a flexible hinge allowing the horizontal alignment of the crystalline cellulose fibrils relative to the cell wall. PMID:24722535

Characterization of Microstructure and Mechanical Properties of Mg-8Li-3Al-1Y Alloy Subjected to Different Rolling Processes

NASA Astrophysics Data System (ADS)

Zhou, Xiao; Liu, Qiang; Liu, Ruirui; Zhou, Haitao

2018-06-01

The mechanical properties and microstructure evolution of Mg-8Li-3Al-1Y alloy undergoing different rolling processes were systematically investigated. X-ray diffraction, optical microscope, scanning electron microscopy, transmission electron microscopy as well as electron backscattered diffraction were used for tracking the microstructure evolution. Tensile testing was employed to characterize the mechanical properties. After hot rolling, the MgLi2Al precipitated in β-Li matrix due to the transformation reaction: β-Li → β-Li + MgLi2Al + α-Mg. As for the alloy subjected to annealed hot rolling, β-Li phase was clearly recrystallized while recrystallization rarely occurred in α-Mg phase. With regard to the microstructure undergoing cold rolling, plenty of dislocations and dislocation walls were easily observed. In addition, the microstructure of alloys subjected to annealed cold rolling revealed the formation of new fresh α-Mg grains in β-Li phase due to the precipitation reaction. The mechanical properties and fracture modes of Mg-8Li-3Al-1Y alloys can be effectively tuned by different rolling processes.

A model for microbially induced precipitation of vadose-zone calcites in fractures at LOS Alamos, New Mexico, USA

NASA Astrophysics Data System (ADS)

Newman, Brent D.; Campbell, Andrew R.; Norman, David I.; Ringelberg, David B.

1997-05-01

Fractures are unique environments that can concentrate the flow of water, nutrients, and contaminants. As such, fractures play an important role in controlling the flux of various substances into and through the vadose zone. Calcite fracture fillings are present in the near surface in the Bandelier Tuff Formation at Los Alamos, New Mexico, and provide a record of the geochemical and hydrologic processes that have occurred in fractures. The objective of this study was to examine calcite fracture fills in order to improve understanding of processes within fractures, and in particular those that lead to precipitation of calcite. Samples of calcite fillings were collected from vertical and horizontal fractures exposed in a shallow waste-burial pit. Scanning electron microscopy show morphologies which suggest that plants, fungi, and bacteria were important in the precipitation process. Quadrupole mass spectrometric analyses of fluid inclusion gases show predominantly methane (17-99%) and little to no oxygen (0-8%), suggesting the development of anaerobic conditions in the fractures. Ester-linked phospholipid biomarkers are evidence for a diverse microbial community in the fractures, and the presence of di-ether lipids indicate that the methane was generated by anaerobic bacteria. The calcite fillings apparently resulted from multiple biological and chemical processes in which plant roots in the fractures were converted to calcite. Roots grew into the fractures, eventually died, and were decomposed by bacteria and fungi. Anaerobic gases were generated from encapsulated organic material within the calcite via microbial decomposition, or were generated by microbes simultaneously with calcite precipitation. It is likely that the biological controls on calcite formation that occurred in the Los Alamos fractures also occurs in soils, and may explain the occurrence of other types of pedogenic calcites.

Biomechanical investigation of impact induced rib fractures of a porcine infant surrogate model.

PubMed

Blackburne, William B; Waddell, J Neil; Swain, Michael V; Alves de Sousa, Ricardo J; Kieser, Jules A

2016-09-01

This study investigated the structural, biomechanical and fractographic features of rib fractures in a piglet model, to test the hypothesis that fist impact, apart from thoracic squeezing, may result in lateral costal fractures as observed in abused infants. A mechanical fist with an accelerometer was constructed and fixed to a custom jig. Twenty stillborn piglets in the supine position were impacted on the thoracic cage. The resultant force versus time curves from the accelerometer data showed a number of steps indicative of rib fracture. The correlation between impact force and number of fractures was statistically significant (Pearson׳s r=0.528). Of the fractures visualized, 15 completely pierced the parietal pleura of the thoracic wall, and 5 had butterfly fracture patterning. Scanning electron microscopy showed complete bone fractures, at the zone of impact, were normal to the axis of the ribs. Incomplete vertical fractures, with bifurcation, occurred on the periphery of the contact zone. This work suggests the mechanism of rib failure during a fist impact is typical of the transverse fracture pattern in the anterolateral region associated with cases of non-accidental rib injury. The impact events investigated have a velocity of ~2-3m/s, approximately 2×10(4) times faster than previous quasi-static axial and bending tests. While squeezing the infantile may induce buckle fractures in the anterior as well as posterior region of the highly flexible bones, a fist punch impact event may result in anterolateral transverse fractures. Hence, these findings suggest that the presence of anterolateral rib fractures may result from impact rather than manual compression. Copyright © 2016 Elsevier Ltd. All rights reserved.

The correlation between acoustic and magnetic properties in the long working metal boiler drum with the parameters of the electron microscope

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ababkov, Nikolai, E-mail: n.ababkov@rambler.ru; Smirnov, Alexander, E-mail: galvas.kem@gmail.com

The present paper presents comparative analysis of measurement results of acoustic and magnetic properties in long working metal of boiler drums and the results obtained by methods of electronic microscopy. The structure of the metal sample from the fracture zone to the base metal (metal working sample long) and the center of the base metal before welding (weld metal sample) was investigated by electron microscopy. Studies performed by spectral acoustic, magnetic noise and electron microscopic methods were conducted on the same plots and the same samples of long working and weld metal of high-pressure boiler drums. The analysis of researchmore » results showed high sensitivity of spectral-acoustic and magnetic-noise methods to definition changes of microstructure parameters. Practical application of spectral-acoustic and magnetic noise NDT method is possible for the detection of irregularities and changes in structural and phase state of the long working and weld metal of boiler drums, made of a special molybdenum steel (such as 20M). The above technique can be used to evaluate the structure and physical-mechanical properties of the long working metal of boiler drums in the energy sector.« less

In vitro and in vivo evaluation of a sublingual fentanyl wafer formulation

PubMed Central

Lim, Stephen CB; Paech, Michael J; Sunderland, Bruce; Liu, Yandi

2013-01-01

Background The objective of this study was to prepare a novel fentanyl wafer formulation by a freeze-drying method, and to evaluate its in vitro and in vivo release characteristics, including its bioavailability via the sublingual route. Methods The wafer formulation was prepared by freeze-drying an aqueous dispersion of fentanyl containing sodium carboxymethylcellulose and amylogum as matrix formers. Uniformity of weight, friability, and dissolution testing of the fentanyl wafer was achieved using standard methods, and the residual moisture content was measured. The fentanyl wafer was also examined using scanning electron microscopy and x-ray diffraction. The absolute bioavailability of the fentanyl wafer was evaluated in 11 opioid-naïve adult female patients using a randomized crossover design. Results In vitro release showed that almost 90% of the fentanyl dissolved in one minute. In vivo, the first detectable plasma fentanyl concentration was observed after 3.5 minutes and the peak plasma concentration between 61.5 and 67 minutes. The median absolute bioavailability was 53.0%. Conclusion These results indicate that this wafer has potential as an alternative sublingual fentanyl formulation. PMID:23596347

A comparative evaluation of dental luting cements by fracture toughness tests and fractography.

PubMed

Ryan, A K; Orr, J F; Mitchell, C A

2001-01-01

In recent years there has been a shift from traditional methods of investigating dental materials to a fracture mechanics approach. Fracture toughness (KIC) is an intrinsic material property which can be considered to be a measure of a material's resistance to crack propagation. Glass-ionomer cements are biocompatible and bioactive dental restorative materials, but they suffer from poor fracture toughness and are extremely susceptible to dehydration. The main objective of this study was to evaluate the fracture toughness of three types of commercially available dental cements (polyacid-modified composite resin, resin-modified and conventional glass ionomer) using a short-rod chevron-notch test and to investigate and interpret the results by means of fractography using scanning electron microscopy. Ten specimens of each cement were fabricated according to manufacturers' instructions, coated in varnish, and stored at ambient laboratory humidity, 100 per cent relative humidity, or in water at 37 degrees C for 7 days prior to preparation for testing. Results indicated that significant differences existed between each group of materials and that the fracture toughness ranged from 0.27 to 0.72 MN/m3/2. It was concluded that the resin-modified glass-ionomer cement demonstrated the highest resistance to crack propagation. Fractographs clearly showed areas of stable and unstable crack growth along the fractured surfaces for the three materials examined.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chan, Shih-Ching; Lo, Shih-Yen; Graduate Institute of Medical Sciences, Tzu Chi University, Hualien, Taiwan

Research highlights: {yields} Lipid rafts are known to play an important role in virus entry and virus assembly of many viruses. {yields} However, HCV is the first example of the association of lipid raft with viral RNA replication. {yields} Our results in this manuscript demonstrate that purified HCV RCs with associated lipid raft membrane appeared as distinct particles of around 0.7 um under EM and AFM. {yields} Knockdown of proteins associated with lipid raft suppressed the HCV replication and reduced the number of these particles. {yields} To our knowledge, structures of HCV RCs were demonstrated at its first time inmore » this manuscript. -- Abstract: Hepatitis C viral RNA synthesis has been demonstrated to occur on a lipid raft membrane structure. Lipid raft membrane fraction purified by membrane flotation analysis was observed using transmission electron microscopy and atomic force microscopy. Particles around 0.7 um in size were found in lipid raft membrane fraction purified from hepatitis C virus (HCV) replicon but not their parental HuH7 cells. HCV NS5A protein was associated with these specialized particles. After several cycles of freezing-thawing, these particles would fuse into larger sizes up to 10 um. Knockdown of seven proteins associated with lipid raft (VAPA, COPG, RAB18, COMT, CDC42, DPP4, and KDELR2) of HCV replicon cells reduced the observed number of these particles and suppressed the HCV replication. Results in this study indicated that HCV replication complexes with associated lipid raft membrane form distinct particle structures of around 0.7 um as observed from transmission electron microscopy and atomic force microscopy.« less

Developing 3D SEM in a broad biological context

PubMed Central

Kremer, A; Lippens, S; Bartunkova, S; Asselbergh, B; Blanpain, C; Fendrych, M; Goossens, A; Holt, M; Janssens, S; Krols, M; Larsimont, J-C; Mc Guire, C; Nowack, MK; Saelens, X; Schertel, A; Schepens, B; Slezak, M; Timmerman, V; Theunis, C; Van Brempt, R; Visser, Y; GuÉRin, CJ

2015-01-01

When electron microscopy (EM) was introduced in the 1930s it gave scientists their first look into the nanoworld of cells. Over the last 80 years EM has vastly increased our understanding of the complex cellular structures that underlie the diverse functions that cells need to maintain life. One drawback that has been difficult to overcome was the inherent lack of volume information, mainly due to the limit on the thickness of sections that could be viewed in a transmission electron microscope (TEM). For many years scientists struggled to achieve three-dimensional (3D) EM using serial section reconstructions, TEM tomography, and scanning EM (SEM) techniques such as freeze-fracture. Although each technique yielded some special information, they required a significant amount of time and specialist expertise to obtain even a very small 3D EM dataset. Almost 20 years ago scientists began to exploit SEMs to image blocks of embedded tissues and perform serial sectioning of these tissues inside the SEM chamber. Using first focused ion beams (FIB) and subsequently robotic ultramicrotomes (serial block-face, SBF-SEM) microscopists were able to collect large volumes of 3D EM information at resolutions that could address many important biological questions, and do so in an efficient manner. We present here some examples of 3D EM taken from the many diverse specimens that have been imaged in our core facility. We propose that the next major step forward will be to efficiently correlate functional information obtained using light microscopy (LM) with 3D EM datasets to more completely investigate the important links between cell structures and their functions. Lay Description Life happens in three dimensions. For many years, first light, and then EM struggled to image the smallest parts of cells in 3D. With recent advances in technology and corresponding improvements in computing, scientists can now see the 3D world of the cell at the nanoscale. In this paper we present the results of high resolution 3D imaging in a number of diverse cells and tissues from multiple species. 3D reconstructions of cell structures often revealed them to be significantly more complex when compared to extrapolations made from 2D studies. Correlating functional 3D LM studies with 3D EM results opens up the possibility of making new strides in our understanding of how cell structure is connected to cell function. PMID:25623622

Microstructure and mechanical properties of bulk and plasma-sprayed y2O3-partially stabilized zirconia

NASA Technical Reports Server (NTRS)

Valentine, P. G.; Maier, R. D.

1980-01-01

Bulk 8.0 weight percent yttria partially stabilied zirconia (PSZ) was studied by light microscopy, transmission electron microscopy, X-ray analysis, microhardness testing, and fracture toughness testing. The as received PSZ contained spheroidal and grain boundary precipitates up to 4 micrometers in size. Spheroids up to 1.26 micrometers were metastable tetragonal; large spheroids were monoclinic. Grinding the PSZ into powder did not cause a significant amount of tetragonal to transform to monoclinic. This indicates that transformation toughness is not a significant mechanism in PSZ. Aging the PSZ at 1500 C caused the fine tetragonal precipitates to grow from 0.06 to 0.12 micrometers, in 250 minutes. A peak hardness of 1400 kg/sq mm was attained after 50 minutes. Solution annealing and quenching the as received PSZ eliminated the large precipitates, but fine tetragonal precipitates reformed on quenching. Aging at 1500 C caused the fine 0.02 micrometers tetragonal precipitates to grow into plates about 0.10 by 0.50 micrometers. A peak hardness of 1517 kg/sq mm was obtained after 250 minutes. On further aging, monoclinic percipitates formed along grain boundaries. The fracture toughness of the aged and unaged solution annealed and quenched PSZ was found to be between 2 and 3 MN /square root of m cubed. This range of fracture toughness is consistent with PSZ's that do not undergo transformation toughening.

Rime and graupel: Description and characterization as revealed by low-temperature scanning electron microscopy

USGS Publications Warehouse

Rango, A.; Foster, J.; Josberger, E.G.; Erbe, E.F.; Pooley, C.; Wergin, W.P.

2003-01-01

Snow crystals, which form by vapor deposition, occasionally come in contact with supercooled cloud droplets during their formation and descent. When this occurs, the droplets adhere and freeze to the snow crystals in a process known as accretion. During the early stages of accretion, discrete snow crystals exhibiting frozen cloud droplets are referred to as rime. If this process continues, the snow crystal may become completely engulfed in frozen cloud droplets. The resulting particle is known as graupel. Light microscopic investigations have studied rime and graupel for nearly 100 years. However, the limiting resolution and depth of field associated with the light microscope have prevented detailed descriptions of the microscopic cloud droplets and the three-dimensional topography of the rime and graupel particles. This study uses low-temperature scanning electron microscopy to characterize the frozen precipitates that are commonly known as rime and graupel. Rime, consisting of frozen cloud droplets, is observed on all types of snow crystals including needles, columns, plates, and dendrites. The droplets, which vary in size from 10 to 100 μm, frequently accumulate along one face of a single snow crystal, but are found more randomly distributed on aggregations consisting of two or more snow crystals (snowflakes). The early stages of riming are characterized by the presence of frozen cloud droplets that appear as a layer of flattened hemispheres on the surface of the snow crystal. As this process continues, the cloud droplets appear more sinuous and elongate as they contact and freeze to the rimed crystals. The advanced stages of this process result in graupel, a particle 1 to 3 mm across, composed of hundreds of frozen cloud droplets interspersed with considerable air spaces; the original snow crystal is no longer discernible. This study increases our knowledge about the process and characteristics of riming and suggests that the initial appearance of the flattened hemispheres may result from impact of the leading face of the snow crystal with cloud droplets. The elongated and sinuous configurations of frozen cloud droplets that are encountered on the more advanced stages suggest that aerodynamic forces propel cloud droplets to the trailing face of the descending crystal where they make contact and freeze.

Study of mixed mode fracture toughness and fracture trajectories in gypsum interlayers in corrosive environment

PubMed Central

Xiankai, Bao; Jinchang, Zhao

2018-01-01

Based on the engineering background of water dissolving mining for hydrocarbon storage in multi-laminated salt stratum, the mixed mode fracture toughness and fracture trajectory of gypsum interlayers soaked in half-saturated brine at various temperatures (20°C, 50°C and 80°C) were studied by using CSNBD (centrally straight-notched Brazilian disc) specimens with required inclination angles (0°, 7°, 15°, 22°, 30°, 45°, 60°, 75°, 90°) and SEM (scanning electron microscopy). The results showed: (i) The fracture load of gypsum specimens first decreased then increased with increasing inclination angle, due to the effect of friction coefficient. When soaked in brine, the fracture toughness of gypsum specimens gradually decreased with increasing brine temperature. (ii) When soaked in brine, the crystal boundaries of gypsum separated and became clearer, and the boundaries became more open between the crystals with increasing brine temperature. Besides, tensile micro-cracks appeared on the gypsum crystals when soaked in 50°C brine, and the intensity of tensile cracks became more severe when soaking in 80°C brine. (iii) The experimental fracture envelopes derived from the conventional fracture criteria and lay outside these conventional criteria. The experimental fracture envelopes were dependent on the brine temperature and gradually expanded outward as brine temperature increases. (iv) The size of FPZ (fracture process zone) was greatly dependent on the damage degree of materials and gradually increased with increase of brine temperature. The study has important implication for the control of shape and size of salt cavern. PMID:29410841

Identification of Ice Nucleation Active Sites on Feldspar Dust Particles

PubMed Central

2015-01-01

Mineral dusts originating from Earth’s crust are known to be important atmospheric ice nuclei. In agreement with earlier studies, feldspar was found as the most active of the tested natural mineral dusts. Here we investigated in closer detail the reasons for its activity and the difference in the activity of the different feldspars. Conclusions are drawn from scanning electron microscopy, X-ray powder diffraction, infrared spectroscopy, and oil-immersion freezing experiments. K-feldspar showed by far the highest ice nucleation activity. Finally, we give a potential explanation of this effect, finding alkali-metal ions having different hydration shells and thus an influence on the ice nucleation activity of feldspar surfaces. PMID:25584435

Studying the effect of photodynamic therapy (PDT) to enhance healing of femur fractures using polarimetric second-harmonic generation microscopy

NASA Astrophysics Data System (ADS)

Golaraei, Ahmad; Raja, Vaishnavi; Akens, Margarete K.; Wilson, Brian C.; Barzda, Virginijus

2017-07-01

Linear polarization-in, polarization-out second-harmonic generation microscopy was used to study the effect of Photodynamic therapy treatment on enhancing the healing of femur fracture by investigating the ultrastructure of collagen as a major component of bone matrix.

Imaging elemental distribution and ion transport in cultured cells with ion microscopy.

PubMed

Chandra, S; Morrison, G H

1985-06-28

Both elemental distribution and ion transport in cultured cells have been imaged by ion microscopy. Morphological and chemical information was obtained with a spatial resolution of approximately 0.5 micron for sodium, potassium, calcium, and magnesium in freeze-fixed, cryofractured, and freeze-dried normal rat kidney cells and Chinese hamster ovary cells. Ion transport was successfully demonstrated by imaging Na+-K+ fluxes after the inhibition of Na+- and K+ -dependent adenosine triphosphatase with ouabain. This method allows measurements of elemental (isotopic) distribution to be related to cell morphology, thereby providing the means for studying ion distribution and ion transport under different physiological, pathological, and toxicological conditions in cell culture systems.

Investigating the fate of nitroaromatic (TNT) and nitramine (RDX and HMX) explosives in fractured and pristine soils.

PubMed

Douglas, Thomas A; Walsh, Marianne E; McGrath, Christian J; Weiss, Charles A

2009-01-01

Explosives compounds, known toxins, are loaded to soils on military training ranges predominantly during explosives detonation events that likely fracture soil particles. This study was conducted to investigate the fate of explosives compounds in aqueous slurries containing fractured and pristine soil particles. Three soils were crushed with a piston to emulate detonation-induced fracturing. X-ray diffraction, energy-dispersive X-ray spectrometry, gas adsorption surface area measurements, and scanning electron microscopy were used to quantify and image pristine and fractured soil particles. Aqueous batches were prepared by spiking soils with solutions containing 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), octahydro 1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), and 2,4-dinitrotoluene (2,4-DNT). Samples were collected over 92 d and the concentrations of the spiked explosives compounds and TNT transformation products 2-amino-4,6-dinitrotoluene (2ADNT) and 4-amino-2,6-dinitrotoluene (4ADNT) were measured. Our results suggest soil mineralogical and geochemical compositions were not changed during piston-induced fracturing but morphological differences were evident with fractured soils exhibiting more angular surfaces, more fine grained particles, and some microfracturing that is not visible in the pristine samples. TNT, 2,4-DNT, RDX, and HMX exhibited greater analyte loss over time in batch solutions containing fractured soil particles compared to their pristine counterparts. 2ADNT and 4ADNT exhibited greater concentrations in slurries containing pristine soils than in slurries containing fractured soils. Explosives compound transformation is greater in the presence of fractured soil particles than in the presence of pristine soil particles. Our results imply fractured soil particles promote explosive compound transformation and/or explosives compounds have a greater affinity for adsorption to fractured soil particle surfaces.

Characterization of the NEXT Hollow Cathode Inserts After Long-Duration Testing

NASA Technical Reports Server (NTRS)

Mackey, J.; Shastry, R.; Soulas, G.

2017-01-01

Hollow dispenser cathode inserts are a critical element of electric propulsion systems, and should therefore be well understood during long term operation to ensure reliable system performance. This work destructively investigated cathode inserts from the NEXT long-duration test which demonstrated 51,184 hours of high-voltage operation, 918 kg of propellant throughput, and 35.5 MN-s of total impulse. The characterization methods used include scanning electron microscopy with energy dispersive spectroscopy and X-ray diffraction. Microscopy analysis has been performed on fractured surfaces, emission surfaces, and metallographically polished cross-sections of post-test inserts and unused inserts. Impregnate distribution, etch region thickness, impregnate chemical content, emission surface topography, and emission surface phase identification are the primary factors investigated.

Why do nickel-titanium archwires fracture intraorally? Fractographic analysis and failure mechanism of in-vivo fractured wires.

PubMed

Zinelis, Spiros; Eliades, Theodore; Pandis, Nikolaos; Eliades, George; Bourauel, Christoph

2007-07-01

The aim of this study was to characterize intraorally fractured nickel-titanium (Ni-Ti) archwires, determine the type of fracture, assess changes in the alloy's hardness and structure, and propose a mechanism of failure. Eleven Ni-Ti SE 200 and 19 copper-Ni-Ti (both, Ormco, Glendora, Calif) intraorally fractured archwires were collected. The location of fracture (anterior or posterior), wire type, cross section, and period of service before fracture were recorded. The retrieved wires and brand-, type-, and size-matched specimens of unused wires were subjected to scanning electron microscopy to assess the fracture type and morphological variation of fracture site of retrieved specimens, and to Vickers hardness (HV200) testing to investigate the hardness of as-received and in-vivo fractured specimens. Fracture site distribution was statistically analyzed with the chi-square test (alpha = 0.05), whereas the results of the hardness testing were analyzed with 2-way ANOVA with state (control vs in-vivo fractured) and composition (Ni-Ti SE vs copper-Ni-Ti) serving as discriminating variables and the Student-Newman-Keuls test at the 95% confidence level. The fracture site distribution showed a preferential location at the midspan between the premolar and the molar, suggesting that masticatory forces and complex loading during engagement of the wire to the bracket slot and potential intraoral aging might account for fracture incidence. All retrieved wires had the distinct features of brittle fracture without plastic deformation or crack propagation, whereas no increase in hardness was observed for the retrieved specimens. Most fractures sites were in the posterior region of the arch, probably because of the high-magnitude masticatory forces. Brittle fracture without plastic deformation was observed in most Ni-Ti wires regardless of archwire composition. There was no increase in the hardness of the intraorally exposed specimens regardless of wire type. This contradicts previous in-vitro studies and rules out hydrogen embrittlement as the cause of fracture.

Micro structrual characterization and analysis of ball milled silicon carbide

NASA Astrophysics Data System (ADS)

Madhusudan, B. M.; Raju, H. P.; Ghanaraja., S.

2018-04-01

Mechanical alloying has been one of the prominent methods of powder synthesis technique in solid state involving cyclic deformation, cold welding and fracturing of powder particles. Powder particles in this method are subjected to greater mechanical deformation due to the impact of ball-powder-ball and ball-powder-container collisions that occurs during mechanical alloying. Strain hardening and fracture of particles decreases the size of the particles and creates new surfaces. The objective of this Present work is to use ball milling of SiC powder for different duration of 5, 10, 15 and 20 hours by High energy planetary ball milling machine and to evaluate the effect of ball milling on SiC powder. Micro structural Studies using Scanning Electron Microscopy (SEM), X-ray Diffraction (XRD) and EDAX has been investigated.

Evaluating stress corrosion cracking behaviour of high strength AA7075-T651 aluminium alloy

NASA Astrophysics Data System (ADS)

Prabhuraj, P.; Rajakumar, S.; Lakshminarayanan, A. K.; Balasubramanian, V.

2017-12-01

The objective of the present study is to determine the threshold stress level of stress corrosion cracking (SCC) in AA7075-T651 aluminium alloy by suitable experimentation. The test was carried out using a circumferential notch specimen in a horizontal-type constant load SCC setup in a 3.5 wt.% NaCl solution. The time to failure by SCC was determined at various loading conditions. The threshold stress of AA7075-T651 alloy was found to be 242 MPa in a 3.5 wt.% NaCl solution. The various regions of the fractured surface specimen such as machined notch, SCC region and final overload fracture area were examined using scanning electron microscopy (SEM) in order to identify the SCC mechanism.

Search for Past Life on Mars: Possible Relict Biogenic Activity in Martian Meteorite ALH84001

NASA Technical Reports Server (NTRS)

McKay, David S.; Gibson, Everett K., Jr.; Thomas-Keprta, Kathie L.; Vali, Hojatollah; Romanek, Christopher S.; Clemett, Simon J.; Chillier, Xavier D. F.; Maechling, Claude R.; Zare, Richard N.

1996-01-01

Fresh fracture surfaces of the martian meteorite ALH84001 contain abundant polycyclic aromatic hydrocarbons (PAHs). These fresh fracture surfaces also display carbonate globules. Contamination studies suggest the PAHs are indigenous to the meteorite. High resolution scanning and transmission electron microscopy study of surface textures and internal structures of selected carbonate globules show that the globules contain fine-grained, secondary phases of single-domain magnetite and Fe-monosulfides. The carbonate globules are similar in texture and size to some terrestrial bacterially induced carbonate precipitates. Although inorganic formation is possible, formation of the globules by biogenic processes could explain many of the observed features including the PAHs. The PAHs, the carbonate globules, and their associated secondary mineral phases and textures could thus be fossil remains of a past martian biota.

Tensile behavior of cenosphere/epoxy syntactic foams

NASA Astrophysics Data System (ADS)

Shahapurkar, Kiran; Doddamani, Mrityunjay; Kumar, G. C. Mohan

2018-04-01

Tensile behavior of syntactic foam composites are very critical to the engineering applications. The fracture modes and failure mechanisms under tension must be fully understood in order to realize the potential of such composites. In the present work, syntactic foam composites are fabricated using as received and surface modified hollow cenospheres embedded into epoxy matrix. Combinations of cenosphere volume fraction (0, 20, 40 and 60%) and surface modification are studied. Experimental results reveal that modulus of both untreated and treated syntactic foams increases with increase in cenosphere volume fraction compared to neat resin. Strength values of syntactic foams show decreasing trend compared to neat resin. However, treated syntactic foams demonstrated better results compared to untreated ones attributing to good bonding between matrix and filler. Scanning electron microscopy reveal brittle fracture for all the syntactic foams.

The effect of aluminum nanoparticles on the structure, mechanical properties and failure of aluminum processed by accumulative roll bonding

NASA Astrophysics Data System (ADS)

Ivanov, K. V.; Fortuna, S. V.; Kalashnikova, T. A.; Rodkevich, N. G.

2017-12-01

The microstructure, mechanical properties, and fracture type of aluminum with and without aluminum nanoparticles processed by accumulative roll bonding (ARB) have been studied using transmission and scanning electron microscopy, microhardness measurements, and tensile tests. It is shown that the injection of aluminum nanoparticles increases the structure refinement rate during ARB due to the increasing tendency for dynamic recrystallization. It has a different effect on different mechanical characteristics. The different effect of nanoparticles on different structural features is the reason for the different effect on different mechanical properties related with these features. The fracture mechanism is shown to change from ductile in aluminum to mixed ductile-brittle in the composite with a 1.5-fold decrease in ductility as a result of nanoparticle injection.

Extended Pulse-Powered Humidity-Freeze Cycling for Testing Module-Level Power Electronics

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hacke, Peter L; Rodriguez, Miguel; Kempe, Michael D

An EMI suppression capacitor (polypropylene film type) failed by 'popcorning' due to vapor outgassing in pulse powered humidity-freeze cycles. No shorts or shunts could be detected despite mildly corroded metallization visible in the failed capacitor. Humidity-freeze cycling is optimized to break into moisture barriers. However, further studies will be required on additional module level power electronic (MLPE) devices to optimize the stress testing for condensation to precipitate any weakness to short circuiting and other humidity/bias failure modes.

A reply to comment on "An experimental study of symmetry lowering of analcime"

NASA Astrophysics Data System (ADS)

Kyono, Atsushi

2018-04-01

The fracture surfaces of analcimes reported by Sugano and Kyono (Phys Chem Miner, 2018) were re-investigated using field-emission scanning electron microscopy. The results of scanning electron microscopic observation show the fractures can be characterized by smoothly curved surfaces, called sub-conchoidal fracture, and neither the lamellar twin nor the domain walls of twin were found on the surfaces of the analcimes. In addition, the lamellar twin is generally formed by transformation from the high-temperature phase or from pseudomorphic replacement under strong alkaline conditions. Actually, the symmetry lowering of analcime reported by Sugano and Kyono (2018) occurs at 200 °C. The temperature is much lower than the transformation from the high-temperature phase. In the hydrothermal experiment (Sugano and Kyono 2018), moreover, the analcimes were synthesized under acidic condition and reheated in pure water. No twin domain is likely to be formed under the hydrothermal condition. These facts strongly deny the possibility that the twin domains cause the pseudo-symmetry of tetragonal analcime. Consequently, it can be concluded that the observed forbidden reflections for the cubic Ia3 d symmetry are not due to the presence of twin domains, but due to the symmetry lowering of analcime from cubic Ia3 d to orthorhombic Ibca.

Effects of deformation-induced martensite and grain size on ductile-to-brittle transition behavior of austenitic 18Cr-10Mn-N stainless steels

NASA Astrophysics Data System (ADS)

Hwang, Byoungchul; Lee, Tae-Ho; Kim, Sung-Joon

2010-12-01

Effects of deformation-induced martensite and grain size on ductile-to-brittle transition behavior of austenitic 18Cr-10Mn-(0.3˜0.6)N stainless steels with different alloying elements were investigated by means of Charpy impact tests and microstructural analyses. The steels all exhibited ductile-to-brittle transition behavior due to unusual brittle fracture at low temperatures despite having a face-centered cubic structure. The ductileto-brittle transition temperature (DBTT) obtained from Chapry impact tests did not coincide with that predicted by an empirical equation depending on N content in austenitic Cr-Mn-N stainless steels. Furthermore, a decrease of grain size was not effective in terms of lowering DBTT. Electron back-scattered diffraction and transmission electron microscopy analyses of the cross-sectional area of the fracture surface showed that some austenites with lower stability could be transformed to α'-martensite by localized plastic deformation near the fracture surface. Based on these results, it was suggested that when austenitic 18Cr-10Mn-N stainless steels have limited Ni, Mo, and N content, the deterioration of austenite stability promotes the formation of deformation-induced martensite and thus increases DBTT by substantially decreasing low-temperature toughness.

Tensile properties of candidate structural materials for high power spallation sources at high helium contents

NASA Astrophysics Data System (ADS)

Jung, P.; Henry, J.; Chen, J.

2005-08-01

Low activation 9%Cr martensitic steels EUROFER97, pure tantalum, and low carbon austenitic stainless steel 316L were homogeneously implanted with helium to concentrations up to 5000 appm at temperatures from 70 °C to 400 °C. The specimens were tensile tested at room temperature and at the respective implantation temperatures. In all materials the helium caused an increased in strength and reduction in ductility, with both changes being generally larger at lower implantation and testing temperatures. After implantation some work hardening was retained in 316L and in tantalum, while it almost completely disappeared in EUROFER97. After tensile testing, fracture surfaces were analysed by scanning electron microscopy (SEM). Implantation caused reduction of necking, but up to concentrations of 2500 appm He fracture surface still showed transgranular ductile appearance. Completely brittle intergranular fracture was observed in tantalum at 9000 appm He and is also expected for EUROFER97 at this concentration, according to previous results on similar 9%Cr steels.