Platinum/iridium/carbon: a high-resolution shadowing material for TEM,STM and SEM of biological macromolecular structures

Thin Pt/Ir/C coating films (1.5 nm) show a fine granularity and provide a high structural resolution in the transmission electron microscope (TEM) when applied to freeze-dried biological macromolecules. They keep their structure when exposed to atmospheric conditions, without the need of an additional stabilizing carbon layer, in contrast to conventional high-resolution shadowing materials such as Ta/W and Pt/C. However, the correct ratio of the components has turned out to be crucial. When evaporating Pt/Ir/C from the source electrode in an electron-beam-heated evaporator, the ratio of the three elements changes progressively, and, consequently, the properties of such films depend strongly on the mass that has been pre-evaporated. In this paper we present a quantitative analysis of the composition of Pt/Ir/C films by wavelength-dispersive X-ray analysis (WDX) undertaken in association with TEM experiments. We applied Pt/Ir/C shadowing to two regular biological test specimens, the phage T4 type III polyhead and the HPI-layer of Deinococcus radiodurans. It turns out that Pt/Ir/C films containing at least 25% C are three-dimensionally stable on the freeze-dried macromolecular samples. By the dramatically improved resolution power of the latest scanning electron microscopes (SEM) and the invention of the scanning tunnelling microscope (STM), two new surface-sensitive tools for the investigation of biological macromolecular structures became available. The Pt/Ir/C coating has proved to be well suited for STM and SEM imaging of freeze-dried biological structures because of its good electrical conductivity and its direct three-dimensional stability. We compare STM, SEM and TEM images of freeze-dried and Pt/Ir/C-coated polyheads.     

Cryo-electron microscopy of helical particles TMV and T4 polyheads

Vitrified Tobacco Mosaic Virus (TMV) and T4 polyhead suspensions have been studied by cryo-electron microscopy. The order of TMV particles is preserved to distances better than 0.3 nm when vitrified. In spite of the high beam sensitivity of unstained biological material, the resolution of images extends to 1.15 nm. Radial density distributions calculated from images of TMV and the helical aggregate of TMV protein (TMVP) show that the RNA is likely visualized in the virus. However, due to the imaging conditions of unstained vitrified specimens, the position of the RNA is erroneous. While TMV and TMVP do not show any departure from cylindrical symmetry, T4 polyheads are sensitive to the thickness of the ice layer in which they are embedded. To avoid flattening, T4 polyheads have to be embedded in an ice layer thicker than their diameter.     

Biological structures imaged in a hybrid scanning transmission electron microscope and scanning tunneling microscope

A hybrid scanning transmission electron microscope (STEM) and scanning tunneling microscope (STM) is described which allows simultaneous imaging of biological structures adsorbed to electron-transparent specimen supports in both modes of scanning microscopy, as demonstrated on uncoated phage T4 polyheads. We further discuss the reproducibility and validity of height data obtained from STM topographs of biomacromolecules and present raw data from topographs of freeze-dried, metal-coated nuclear envelopes from Xenopus laevis oocytes.     

Inspection of the adhesion strength of an electrical insulating coating with the surface of anisotropic steel

It has been shown that the thickness of a coating does not affect its adhesion strength with metal. As the metal thickness increases, for each 0.01 mm, the critical diameter of bending of a specimen at which the coating exfoliates increases by 1 mm. The presence of chromium in the electrical insulating coating causes its adhesion properties to decrease. The quality of the ground layer is a very important characteristic affecting the adhesion strength between the coating and metal.     

Correlation of surface topography of metal-shadowed specimens with their negatively stained reconstructions

We present a comparison of surface reconstructions from three different freeze-dried and unidirectionally metal-shadowed specimens (i.e. bacteriophage T4 polyheads, crystalline actin sheets and nuclear pore complexes) with two- or three-dimensional reconstructions of the same specimens when prepared by negative staining. Based on these and many published results, the following conclusions have been reached: With a "cooperative" specimen (e.g. the polyheads), the surface reconstruction computed from a metal-shadowed replica compares favorably with two- or three-dimensional reconstructions obtained from the same specimen after negative staining at the 3-4 nm resolution level. This relatively "poor" level at which the surface topographies of the two preparations can be compared appears to be set by a "practical" resolution limit (i.e. of distinct and reproducible structural detail) of metal replicas of biological specimens, despite the appearance of weak higher-order diffraction spots (i.e. corresponding to 2-3 nm). While in some cases (e.g. the crystalline actin sheets) the surface reliefs of metal replicas may bear little resemblance to the actual structure under investigation, the replicas may still contain sufficient features to establish the polarity or handedness of the structure (i.e., the "top" and "bottom" surfaces of a crystalline sheet). Information from negatively stained specimens is usually complementary with information from freeze-dried and metal-shadowed specimens. However, there are artifacts in both techniques, and we present an example with the nuclear pore complex, where these techniques yield confusing results.     

铀表面铝镀层热应力的有限元分析

对铀表面磁控溅射沉积铝镀层的热应力进行了热弹塑性有限元分析。结果表明：镀层内的热应力较大,达到铝的屈服强度,镀层界面两侧存在明显应力梯度,试样侧边存在由于边缘效应引起的应力分布不均匀性,至侧边约4倍镀层厚度处,应力分布不均匀性逐渐消失;沉积温度升高,界面塑性应变明显增大,镀层弹性模量和泊松比对镀层界面热应力和塑性应变的影响较小,而屈服强度的影响较大,减薄镀层厚度有利于改善镀层界面的热应力和塑性应变。     

近疲劳强度载荷下WC/Co涂层对300M钢疲劳性能的影响

研究了在近疲劳强度载荷下超音速火焰喷涂WC/Co涂层和电镀铬层对300 M低合金钢疲劳性能的影响.结果表明：WC/Co涂层、电镀铬层试样和300 M钢基体的疲劳强度分别为753,600和720 MPa.在近疲劳强度载荷下,WC/Co涂层试样的疲劳寿命分别为基体和电镀铬层试样的1.5～2和5～10倍.三者的主裂纹源均为基体内部的Al2O3和SiO2夹杂物.电镀铬层中的次裂纹穿过界面并向基体中扩展,在基体表面形成新的次裂纹源,加快了基体中主裂纹源的形成和主裂纹的扩展;而WC/Co涂层中的次裂纹经过界面时向界面方向偏转,形成无支撑涂层,并最终使涂层剥落.     

Development of coatings based on Fe-Cr-Ni-Al for resisting fretting wear in the temperature range 20–700 °C

Previous work by the present workers has indicated that sprayed molybdenum coatings on steel were beneficial in resisting fretting wear in the temperature range 20–300 °C. Beyond this temperature range the oxidation rate of molybdenum increased rapidly and limited its use. Such coatings were mixtures of metal and oxide and provided an elevated temperature “glaze” which appeared to lubricate the surface and to reduce significantly the level of fretting damage. In developing a coating for use over a wider temperature range, mixtures of elements were selected which would provide the possibility of surface glaze formation.Mixtures of Fe-Cr and Ni-Al alloys were arc sprayed onto a low alloy steel and tested under conditions of fretting at 20, 475 and 700 °C. Coatings containing iron and chromium developed a glaze oxide at 475 and 700 °C with resultant low wear. The nickel-based coating did not develop the glaze oxide at 475 °C but did so at 700 °C.     

Electron probe x-ray microanalysis of frozen-hydrated biological specimens.

A technique is described for preparing frozen-hydrated bulk samples of biological specimens for electron probe X-ray microanalysis. The method allows reproducible quantitative analyses to be made. Specimens are rapidly frozen, transferred to a vacuum evaporator, fractured under high vacuum at - 180 degrees C and coated with 20 nm of chromium. Transferal to the cryostage of a scanning electron microscope is accomplished without exposure to the atmosphere and without the specimen temperature rising above -120 degrees C. Analyses are made at a temperature of -145 degrees C. Contamination by frost does not occur. Etching and charging of the specimen are eliminated. Specimen charging is shown to be related to temperature. It can be eliminated at low temperature by coating with carbon, aluminium or chromium but consistent elimination could only be achieved with chromium. The chromium coat does not appear to have an absorption effect on quantitative analysis.     

Durability of chromium plating under dry sliding conditions

Experimental data are presented to show that engineering chromium plating on cast iron fails catastrophically after a given number of sliding cycles depending upon the applied load and the coating thickness. The failure mechanism involves the initiation of cracks in the chromium both at its sliding surface and at the coating-substrate interface adjacent to graphite flakes. The cracks propagate through the coating to give fracture and mechanical breakdown of the coating. The use of durability limit curves to identify ‘fail-safe’ conditions is discussed.     

Sliding tribological behavior of AlCrN coating

The sliding tribological behavior of the PVD AlCrN coating against Si₃N₄ ball have been investigated by using the CETR multi-functional UMT-2 test system under two sliding conditions (bidirectional and unidirectional). Reciprocating sliding tests (bidirectional) were performed under varied normal loads (5, 10 and 20 N) at sliding velocity of 0.48 m/min. Ball-on-disc tests (unidirectional) were performed at varied sliding velocities (0.48 and 5 m/min) under normal load of 5 N. The wear scars of the coating were evaluated by surface profilometer, scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDX), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS), and the sliding wear mechanism of the coating was consequently discussed. The results showed that AlCrN coating had excellent anti-abrasion properties. Both the normal load in reciprocating sliding test and the sliding velocity in ball-on-disc test had significant influence on the sliding tribological behavior of the AlCrN coating. The combination of abrasion and oxidation was the main sliding wear mechanism for the AlCrN coating. The wear resistant and thermally stable oxides formed by the tribo-chemical reactions of chromium and aluminum protected the AlCrN coating against wear admirably.     

Chromium oxide coatings applied to magnetic tape heads for improved wear resistance

This paper reports on an investigation of the wear of chromium oxide based very thin films. Linear data tape Advanced Digital Recording (ADR™) heads coated with 20- and 40-nm thick chromium oxide films have been tested subject to temperature/humidity matrix of 10 to 40°C/10 to 80% in order to assess the wear behaviour of the coating as a function of environment. The tested heads were analysed at various stages of wear, by use of optical microscopy (OM), atomic force microscopy (AFM) and Auger electron spectroscopy (AES). The results show that the most severe damage occurs at the highest relative humidity (80% RH) and for a given humidity, at the lowest temperature (10°C). Inversely, stain transferred from the tape to the head surface predominates at the lowest humidity (10% RH). Stain therefore appears to protect the coating against wear with the degree of protection increasing with the temperature.The wear process differs according to the coating thickness. This is attributed to the location of the maximum subsurface stress with respect to the coating/substrate interface. It is shown that this maximum stress occurs below the interface for 20 nm and at the interface for 40 nm thick coatings. This correlates to different observed wear modes.     

A procedure for increasing the contrast of biological specimens in edge-projection TEM

Rotary shadowing has been used to increase the image contrast of biological specimens during edge-projection imaging in the transmission electron microscope (TEM). In this imaging mode, biological specimens are adsorbed from aqueous solution onto a highly curved substrate and observed in a direction parallel to its surface. High contrast TEM images are obtained at 200 kV when a 1–3 nm layer of tungsten is thermally evaporated onto the substrate at an angle of about 9°. Individual adsorbates are clearly delineated by the smooth, continuous, and fine-grained tungsten layer that surrounds them. TEM images obtained with this technique can provide a unique view of biological adsorbates on metal, insulator or semiconductor substrates.     

Structure analysis of sputter-coated and ion-beam sputter-coated films: a comparative study

Gold, platinum and tungsten films were deposited by low energy input (7 mA, 450 V), or high deposition rate (80 mA, 1500 V), diode sputter coating and by ion beam sputter coating. Film structures on Formvar coated grids and on the surface of coated erythrocytes, resin embedded, sectioned, and recorded at high magnification in a TEM were compared using computer-assisted measurements and analysis of film thickness and grain size. The average grain size of the thinnest gold and platinum films was relatively independent of the mode or rate of deposition but as the film thickness increased, significant differences in grain size and film structure were observed. Thick platinum or gold films deposited by low energy input sputter coating contained large grain size and electron transparent cracks; however, more even films with narrower cracks but larger grain size were produced at high deposition rates. Ion beam sputter coated gold had relatively large grain size in 10 nm thick films, but beyond this thickness the grains coalesced to form a continuous film. Platinum films deposited by ion beam sputter coating were even and free of electron transparent cracks and had a very small grain size (1–2 nm), which was relatively independent of the film thickness. Tungsten deposition either by low energy input or ion beam sputter coating resulted in fine grained even films which were free of electron transparent cracks. Such films remained granular in substructure and had a grain size of about 1 nm which was relatively independent of film thickness. Tungsten films produced at high deposition rates were of poorer quality. We conclude that thick diode sputter coated platinum and gold films are best deposited at high deposition rates provided the specimens are not heat sensitive, the improvement in film structure being more significant than the slight increase in grain size. Thick diode or ion beam sputter coated gold films should be suitable for low resolution SEM, and thin discontinuous gold films for medium resolution SEM. Diode sputter coated platinum should be suitable for medium resolution SEM and ion beam sputter coated platinum for medium and some high resolution SEM. 1–5 nm thick tungsten films, deposited by low energy input or ion beam sputter coating should be suitable for high resolution SEM, particularly where contrast is of less importance than resolution.     

Particle-surface interactions during the erosion of aluminide-coated MarM002

Particle-surface interactions during the erosion of a nickel aluminide coating were assessed using a single-impact technique. It is shown that the erosive response is a function of the surface scale thickness and the temperature, with the temperature not only influencing the surface scale plasticity but also determining the contribution of the coating substrate to the impact process. In this respect the ductile-to-brittle transition temperature of the coating is of particular importance.Under a wide range of conditions typical of those found in gas turbines the erosion of aluminide coatings is shown to be controlled by the formation and removal of surface scales. This implies that the use of aluminide coatings will increase the erosion resistance of typical turbine blade materials because of the superior oxidation and corrosion resistance of this coating. This increase in erosion resistance will be particularly significant at higher operating temperatures, above 900 °C.     

Reciprocating Sliding Wear Performance of Hard Coating on Modified Titanium Alloy Surfaces

The high strength, low weight, and outstanding corrosion resistance properties possessed by titanium alloys have led to a wide range of successful applications in aerospace, automotive, and chemical industries and in power generation. Titanium alloys are characterized by poor wear resistance properties and their utilization has been excessive in nontribological applications. Surface texturing is a well-known and effective means of surface modification to improve the tribological properties of sliding surfaces. In the present work, modification of titanium alloy surfaces (Ti6Al4V) was done by lapping and laser surface texturing. The wear-resistant coating, AlCrN, was applied over the modified titanium alloy surfaces, with and without a chromium interlayer. Linear reciprocating sliding wear tests were performed with ball-on-flat contact geometry to evaluate the tribological performance of the coated alloy. The tests were performed under different normal loads for a period of 105 cycles at a frequency of 5 Hz. The friction force between the contact pair and displacement of the ball were simultaneously observed using a force transducer and laser displacement sensor. Optical microscopy was used to quantify the wear volume by measuring the wear scar diameter on both the specimen and the counterbody. Scanning electron microscopy (SEM) was employed to study the morphology of the wear scar. The characteristic behavior of the AlCrN coating such as bonding strength, wear volume, wear rate, and coefficient of friction with the chromium interlayer was evaluated and compared with the coating directly applied over the substrate. The coating on the textured surface, with the chromium interlayer showed better tribological performance.     

FDM 3D打印聚醚醚酮零件摩擦磨损特性研究

为研究3D打印各向异性对摩擦性能的影响,通过熔融沉积成型(FDM)制备了0°、45°、90°3种打印角度的聚醚醚酮(PEEK)试样,研究3种不同打印角度及载荷变化对PEEK试样摩擦学性能及磨损机制的影响。利用MFT-5000摩擦磨损试验机对PEEK材料进行室温水润滑下的往复滑动摩擦试验,用超景深显微镜观察磨损后表面形貌。试验结果表明：不同载荷下3种打印角度试样的摩擦因数由大到小依次为0°试样、90°试样、45°试样,磨损率由大到小依次为90°试样、45°试样、0°试样;随着载荷的增大,3种不同打印角度试样的摩擦因数均呈现下降趋势,磨损率则呈现上升趋势;PEEK磨损机制是黏着磨损以及疲劳磨损引起的表层脱落。     

High temperature friction and wear characteristics of various coating materials for steam valve spindle application

Various coatings such as chromium carbide (deposited by plasma spraying and detonation gun techniques), chromium oxide, chromium oxide+titania+silica, NiCrAlY, and Al₂O₃+NiAl, all deposited by plasma spraying; stelliting, and surface nitriding have been applied on X20CrMo V121 steel. This steel is used for high temperature applications such as steam turbine valve spindle. Friction and wear behavior of the surface coated and treated materials have been studied at an elevated temperature of 550°C while rubbing against graphite-filled stellited steel. These studies have been carried out on SRV Optimol reciprocating tribometer. Test parameters for tribological studies have been selected with a view to simulate operating conditions encountered in operation. Additionally, the structure, porosity, hardness, bond strength, and thermal cycling behaviour of these surface coated/treated materials have been characterised. Based on these laboratory investigations, chromium carbide coating deposited by plasma spraying technique has been identified as the most suitable coating for steam turbine valve spindle application. Process parameters have been established for deposition of chromium carbide coating by plasma spraying technique on actual valve spindles. The field results obtained are found to be commensurate with the laboratory findings.     

Freeze-fracture of 3T3 cells for high-resolution scanning electron microscopy

Triton-extracted, freeze-fractured 3T3 cells have been examined in the Hitachi S-900 field-emission SEM, after light platinum coating, at low beam voltage to evaluate the performance of the microscope under these conditions. For unstained material fixed in glutaraldehyde alone, high-resolution images can be obtained, at accelerating voltages of 1.5-5kV, after rotary deposition of platinum to an average thickness of 1.5-3nm. Comparisons are made between these results and those of studies by TEM of deep-etch replicas of similar material previously published.     

Use of sputter coating to prepare whole mounts of cytoskeletons for transmission and high-resolution scanning and scanning transmission electron microscopy

This paper describes the use of sputter coating to prepare detergent-extracted cytoskeletons for observation by scanning (SEM), scanning transmission (STEM), inverted contrast STEM, and transmission (TEM) electron microscopy. Sputtered coats of 1–2 nm of platinum or tungsten provide both an adequate secondary electron signal for SEM and good contrast for STEM and TEM. At the same time, the grain size of the coating is sufficiently fine to be just at (platinum) or below (tungsten) the limit of resolution for SEM and STEM. In TEM, the granular structure of platinum coats is resolved, and platinum decoration artifacts are observed on the surface of structures. The platinum is deposited as small islands with a periodic distribution that may reveal information about the underlying molecular structure. This method produces samples that are similar in appearance to replicas prepared by low-angle rotary shadowing with platinum and carbon. However, the sputter-coating method is easier to use; more widely available to investigators; and compatible with SEM, STEM, and TEM. It may also be combined with immunogold and other labeling methods. While TEM provides the highest resolution images of sputter-coated cytoskeletons, it also damages the specimens owing to heating in the beam. In SEM and STEM cytoskeletons are stable and the resolution is adequate to resolve individual microfilaments. The best single method for visualizing cytoskeletons is inverted contrast STEM, which images both the metal-coated cytoskeletal structures and electron-dense material within the nucleus and cytoplasm as white against a dark background. STEM and TEM were both suitable for visualizing colloidal gold particles in immunolabeled samples.