Hemocyanin - antibody labeling of phodopsin in mouse retina for a scanning electron microscope study.

To reveal the presence of rhodopsin on the surface of the mouse retina, a scanning electron microscope study of the immunolabeling of rhodopsin was attempted. The glutaraldehyde-fixed mouse retina was treated first with rabbit antibodies specific against bovine rhodopsin and then with hemocyanin-labeled goat antibodies specific against rabbit antibody. The distribution of hemocyanin label on mouse retina and the technique used for labeling are discussed.     

X-Ray fluorescence analysis in the scanning electron microscope

Summary X-ray fluorescence analysis can be performed in a scanning electron microscope equipped with an energy dispersive X-ray spectrometer by means of a special attachment. For the production of primary X-rays a target is positioned in the electron beam. With this technique an improvement of the detection limits is obtained because of the small fraction of bremsstrahlung in the X-ray spectrum. As a disadvantage a loss of lateral resolution occurs. The performance features of different types of construction (transmission and massive targets) are be compared for experimental results and theoretical models.The paper has been presented at 12th Colloquium on Materials Analysis, Institute for Analytical Chemistry, Technical University in Vienna, May 13–15, 1985.     

Laser experiments in a scanning electron microscope

In order to increase the information content of electron microscopic investigations a scanning electron microscope was combined with a pulsed Nd-glass laser in both the free running and the Q-switch mode. The equipment allows an in situ observation of melting, sintering and evaporation processes as well as of crack generation and growth at target surfaces. This article describes evaporation experiments for laser PVD of HTSC layers, Langmuir probe measurements in the corresponding plasma and crack processes by thermal shock loading in the SEM.     

Imaging modes for scanning confocal electron microscopy in a double aberration-corrected transmission electron microscope.

Aberration correction leads to reduced focal depth of field in the electron microscope. This reduced depth of field can be exploited to probe specific depths within a sample, a process known as optical sectioning. An electron microscope fitted with aberration correctors for both the pre- and postspecimen optics can be used in a confocal mode that provides improved depth resolution and selectivity over optical sectioning in the scanning transmission electron microscope (STEM). In this article we survey the coherent and incoherent imaging modes that are likely to be used in scanning confocal electron microscopy (SCEM) and provide simple expressions to describe the images that result. Calculations compare the depth response of SCEM to optical sectioning in the STEM. The depth resolution in a crystalline matrix is also explored by performing a Bloch wave calculation for the SCEM geometry in which the pre- and postspecimen optics are defocused away from their confocal conditions.     

An improved visual tracking method in scanning electron microscope

Since their invention, nanomanipulation systems in scanning electron microscopes (SEMs) have provided researchers with an increasing ability to interact with objects at the nanoscale. However, most nanomanipulators that are capable of generating nanometer displacement operate in an open-loop without suitable feedback mechanisms. In this article, a robust and effective tracking framework for visual servoing applications is presented inside an SEM to achieve more precise tracking manipulation and measurement. A subpixel template matching tracking algorithm based on contour models in the SEM has been developed to improve the tracking accuracy. A feed-forward controller is integrated into the control system to improve the response time. Experimental results demonstrate that a subpixel tracking accuracy is realized. Furthermore, the robustness against clutter can be achieved even in a challenging tracking environment.     

Ultramicrohardness tester for use in a scanning electron microscope

Summary A novel microhardness-tester has been designed for loads between 10^–2 N and 5 x 10^–5 N. The apparatus can be operated in a scanning electron microscope without installation of additional adjusting shafts. The force is generated electromagnetically, transduced to the indenter (diamond cone or pyramid) by bending a double leaf spring, and measured by means of strain gauges fixed to the springs. The indentation cycle is fully programmable and is controlled by the strain gauge signal. The equipment can be tilted with respect to the electron beam thus making possible the observation of the tip during indentation.With 2 figures     

Quantitative trace analysis with X-ray fluorescence in the scanning electron microscope

The potential of a scanning electron microscope will be enhanced markedly by adapting the X-ray fluorescence for material analysis. The illuminating system röntgenbox enables the analyst to execute analyses in the concentration range from 100 percent down to the low ppm range without time consuming preparation steps. X-ray intensities of traces are measured for different anodes in the elemental range fromZ=11 to 90 (sodium to thorium).     

A combined ultrahigh vacuum scanning tunneling-scanning electron microscope system

A combined ultrahigh vacuum scanning tunneling-scanning electron microscope system is presented. This system allows to approach arbitrary locations on a sample within a range of 5 mm × 5 mm under electron microscopic control. The usefulness of this system is demonstrated by the characterization of a 1 μm wide photolithographically produced Ag microbridge on a Si(100) substrate.     

蔡司GeminiSEM 300型场发射扫描电镜常见故障的预防、排除和磁性样品的测试方法

金属有机框架材料（MOFs）的形貌结构对其性能应用具有很大影响,但MOFs普遍存在导电性差且对电子束敏感等问题,在进行扫描电子显微镜（SEM）测试时容易损伤样品,发生荷电现象. 因此摸索合适的测试参数,对获得高质量的MOFs扫描电子显微镜图像具有重要意义. 以MIL-101(Cr)、Fe-MOF、Mn-MOF、ZIF-67(Co)这4种典型的MOFs为例,主要探究了加速电压、电子束流、工作距离、探头及喷金对其成像效果的影响. 结果表明,升高加速电压可有效提高图像分辨率,但同时电子束穿入深度增大,可能导致电荷击穿效应对其表面结构造成破坏. 适当增大束流可提高图像信噪比,但过大的束流会导致纳米颗粒边缘变钝,因此选用0.1~0.4 nA中等束流为佳. 在选择探头时需注意,艾弗哈特-索恩利探头（ETD）和透镜内二次电子（T2）探头所成图像立体感较好,透镜内背散射电子（T1）探头的立体感弱,但衬度较好,柱内二次电子（T3）探头分辨率最佳,但更容易荷电,显得颗粒扁平. 而喷金处理可有效提高样品的导电性. 以上结果对使用SEM探究MOFs形貌结构具有一定的借鉴作用.     

Closed-cell foam skin thickness measurement using a scanning electron microscope

Closed cell polymer foam skin thickness can be assessed by taking backscatter electron (BSE) images in a scanning electron microscope (SEM) at a series of accelerating voltages. Under a given set of experimental conditions, the electron beam mostly passes through thin polymer skin cell walls. That cell appears dark compared to adjacent thicker-skinned cells. Higher accelerating voltages lead to a thicker skin being penetrated. Monte Carlo modeling of beam-sample interactions indicates that at 5 keV, skin less than ～0.5 μm in thickness will appear dark, whereas imaging at 30 keV allows skin thicknesses up to ～4 μm to be identified. The distribution of skin thickness can be assessed over square millimeters of foam surface in this manner. Qualitative comparisons of the skin thicknesses of samples can be made with a simple visual inspection of the images. A semiquantitative comparison is possible by applying image analysis. The proposed method is applied to two example foams. Characterizing foam skin thickness by this method is possible using any SEM that is capable of collecting useful BSE images over a range of accelerating voltages. Imaging in low vacuum, where an electrically conductive metal coating is not required, leads to more sensitivity in skin thickness characterization.     

Use of cathode lens in scanning electron microscope for low voltage applications

At a landing energy of 10 eV it is possible to achieve spatial resolution of the same order as at the nominal energy, which is usually 15 keV in the classical scanning electron microscope, by taking advantages of the optical properties of the cathode lens. Two different types of the detection system were designed and tested to learn as much about the optical properties of this system as possible and to start to understand the contrast mechanisms at very low energies. Great changes in the contrast take place when the landing energy is changed from 10 eV to an energy of about 2 keV.     

Mechanical characterization of nanofibers using a nanomanipulator and atomic force microscope cantilever in a scanning electron microscope

The mechanical characterization of polymeric nanofibers is essential to explore the potential for their applications in structural materials, such as nanofiber-reinforced polymer composites. A ‘hooking’ method using a nanomanipulator and atomic force microscope (AFM) cantilever was developed in a scanning electron microscope (SEM). First, a single electrospun nanofiber was suspended over a trench in a substrate. The midpoint of the nanofiber was then hooked by the AFM cantilever. As the AFM cantilever moved in the transverse direction of the nanofiber, the nanofiber was elongated until it fractured. The hooking and elongation processes were controlled by a nanomanipulator. This method features an affordable configuration and loading mechanism, because it does not rely on the firm grip of the nanofiber at both ends. Nanofibers with different diameters were tested using this method, demonstrating that nanofibers with a smaller diameter have greater strength and, thus, are highly suitable for use as a nano-reinforcement in composite applications.     

Charge-related problems associated with X-ray microanalysis in the variable pressure scanning electron microscope at low pressures.

In variable pressure scanning electron microscopy (VPSEM) the current data suggests that considerable caution is required in the interpretation of X-ray data from nonconductive samples, depending on the operating conditions. This article reviews some of the documented approaches and presents data that illustrate the nature and magnitude of the effects of charge above, on, and in the sample on the detected X-ray emissions from the sample and from elsewhere within the VPSEM specimen chamber. The collection of reliable and reproducible X-ray data has been found to require relatively high specimen chamber gas pressures, at the upper end of or beyond the available pressures for most VPSEMs. It is also shown that sample characteristics, including composition, strongly influence local charge effects, which can significantly affect the primary electron landing energy and consequently the resultant emitted X-ray signal under low pressure environments.     

The sub-membrane reticulum of the human erythrocyte: a scanning electron microscope study

A web-like reticulum underlying the human erythrocyte membrane was studied at a resolution of 5-10 nm by means of scanning electron microscope. The network was visualized in isolated membranes (ghosts) torn open to reveal their interior space and in residues derived from ghosts extracted with Triton X-100. It formed a continuous (rather than patchy) cover over the entire cytoplasmic surface, except where lifted off or torn away. Filaments (5-40 nm in diameter), annular figures (40-60 nm in diameter), and nodes (30-100 nm in diameter) were prominent in different networks. The dimensions of the filaments and the interstices in the reticulum varied with conditions, suggesting that the network has elastic properties. This reticulum is probably related to the erythrocyte membrane proteins spectrin and actin.     

A preparation technique for examination of wet-spun polymer fibers in a scanning electron microscope

Summary In this paper the technique is discussed of sample preparation for freshly wet-spun polymer fibers, to be examined by scanning electron microscopy.It makes use of cryogenic breaking of the samples, followed by freeze drying or freeze-etching of the specimen and coating it with a charge conducting layer.The method can also be adapted to the investigation of intermediate coagulation structures of the spinning fiber. Results are discussed for nylon-6 and polyurethane wet-spun fibers.

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Zusammenfassung In diesem Beitrag wird eine Methode zum Präparieren von Proben naß-gesponnener Polymerfasern, die noch nicht weiter bearbeitet sind, zwecks Untersuchung mit einem Raster-Elektronenmikroskop, besprochen.Bei dieser Methode werden die Fasern kryogen gebrochen, gefolgt durch Gefriertrocknen oder Gefrierätzen der Fasern, wonach die Fasern mit einer gut leitenden Schicht überdeckt werden.Diese Methode kann auch angewendet werden für die Untersuchung von Zwischenstrukturen der Fasern während des Spinnprozesses.Ergebnisse werden besprochen für Nylon-6 und Polyurethane-naß-gesponnene Fasern.

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With 8 figures

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Research Associate at Twente University during the academic year 1971–1972.     

Quantitative energy dispersive X-ray analysis of submicrometric particles using a scanning electron microscope

The quantitative scanning electron microscope-energy dispersive X-ray (SEM-EDX) analysis of a horneblende and two augite prismatic samples reduced to submicrometric particles was performed, and error due to the particle effects ("absent mass" and the "reduced absorption" effect) was minimized. Correction factors as a function of fragment size were obtained for O, Na, Mg, Si, Ca, and Fe. In addition, the influence of chemical composition of the samples used as standards (the matrix effect) on correction factors was evaluated. The results indicate that the absent mass effect is dominant for all elements except for the light elements O and Na, for which the reduced absorption effect is dominant. No significant matrix effect has been observed. By using corrected SEM-EDX data, the error on quantification of the element concentration has been estimated to be 3% relative for light elements and below 2% relative for heavy elements (notably, about 1% relative for Fe).     

Transmission electron microscope and scanning auger investigations of temper-embrittled 12% Cr steel

Summary Reversible temper-embrittlement of 12% Cr martensitic steel was investigated. It was observed by SEM that morphology of the carbides found on the smooth intercrystalline fracture facets of the embrittled sample is much different from M₂₃C₆ carbides observed by TEM on the former austenite grain boundaries. Morphology of the carbides on the smooth facets matches TEM data for the M₇C₃ carbides formed during the embrittlement treatment in some distance from the former austenite grain boundaries. Associated SAM analysis data suggest that phosphorus tends to concentrate within stress fields formed around M₇C₃ carbides.

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Untersuchungen von Versprödungseffekten in Cr-Iegierten Stählen mit Transmissionelektronenmikroskopie und Auger-Spektroskopie