Extended abstract of a paper presented at Microscopy and Microanalysis 2004 in Savannah, Georgia, USA, August 1–5, 2004.

Chemically functionalized metal cluster compounds have demonstrated important advantages over colloidal gold as biological microscopy labels. They are covalently cross-linked to the targeting biomolecule, and therefore may be conjugated to a wide range of molecules which cannot be labeled with colloidal gold. The 1.4 nm Nanogold® cluster has been conjugated to peptides, lipids and oligonucleotides, some of which have been proposed as elements of novel molecular wires and novel materials. Dissociation of colloidal gold particles from the conjugate probe, a source of error in quantitative immunogold studies, is greatly reduced by covalent cross-linking. Nanogold® is an uncharged molecule, and because its surface is completely coordinated by organic ligands, nonspecific binding is greatly reduced. Nanogold® conjugates also show greatly enhanced penetration into cells and tissue sections. However, gold probes larger than Nanogold® are desirable for improved visualization in specimens with electron-dense regions or staining, or for applications such as double labeling studies with different sized gold labels, or visualizing wider antigen distributions.

Extended abstract of a paper presented at Microscopy and Microanalysis 2009 in Richmond, Virginia, USA, July 26 – July 30, 2009

Silver nanocrystals passivated by dodecanethiol self-assembled monolayers were produced using an aerosol technique described in detail elsewhere [1]. Self-assembling passivated nanocrystal-superlattices (NCS's) involve self-organization into monolayers, thin films, and superlattices of size-selected nanoclusters encapsulated in a protective compact coating [2,3,4,5,6,7]. We report the preparation and structure characterization of three-dimensional (3-D) hexagonal close-packed Ag nanocrystal supercrystals from Ag nanocrystals of ˜4.5 nm in diameters. The crystallography of the superlattice and atomic core lattices were determined using transmission electron microscopy (TEM) and high-resolution TEM.

SEM was used to image the nanocrystal superlattices formed on an amorphous carbon film of an TEM specimen grid (fig. la). The superlattice films show well shaped, sharply faceted, triangular shaped sheets. Figure lb depicts numerous Ag nanocrystal aggregates uniformly distributed over the imaging region. Inset in this figure is an enlargement of the boxed region at the edge of a supercrystal typifying the ordered nanocrystal packing.

This tutorial will describe the technique of electron backscattered diffraction (EBSD) in the scanning electron microscope (SEM). To properly exploit EBSD in the SEM it is important to understand how these patterns are formed. This discussion will be followed by a description of the hardware required for the collection of electron backscatter patterns (EBSP). We will then discuss the methods used to extract the appropriate crystallographic information from the patterns for orientation determination and phase identification and how these operations can be automated. Following this, a number of applications of the technique for both orientation studies and phase identification will be discussed.

EBSD in the SEM is a phenomenon that has been known for many years. EBSD in the SEM is a technique that permits the crystallography of sub-micron sized regions to be studied from a bulk specimen. These patterns were first observed over 40 years ago, before the development of the SEM and were recorded using a special chamber and photographic film.

The present study describes the development and use of a new bioconjugate combining targeted quantum dot labeling with an immunoperoxidase method and explores whether these bioconjugates could specifically and effectively label Cu/Zn superoxide dismutase (SOD1). The new bioconjugate is designed for the examination of samples both under fluorescent and bright-field microscopy at the same time. For this purpose chlorobis(2-2′-bipyridyl) methacryloyl tyrosine-ruthenium(II) and bis (2-2′-bipyridyl) methacryloyltyrosine-methacryloyltryptophan-ruthenium (II) photosensitive monomers and photosensitive poly(Bis (2-2′-bipyridyl)) methacryloyltyrosine-methacryloyltryptophan-ruthenium(II) were synthesized and characterized. The anti-SOD1 antibody and horseradish peroxidase (HRD) conjugated quantum dots were prepared by using this polymer. The anti-SOD1 antibody and HRD conjugated quantum dots were used in labeling and imaging of SOD1 in rat liver sections. Quantum dot particles were observed as a bright fluorescence in their specific binding locations inside the hepatocytes. The HRD-diaminobenzidine reaction product was observed as brown-colored particles at the same locations under bright-field microscopy. Structural details of the tissue sections could be examined at the same time. The conjugation protocol is simple; the bioconjugate is applicable for efficient cell labeling and can be adapted for imaging of other targets in different tissues. Also, the prepared nanobioconjugates have mechanic stability and can be used for a long period.

The adsorption of polysaccharides onto solid substrates provides an environment for the immobilization of biomolecules [1] giving rise to conditions for the development of biosensors [2] and kits for immunoassays. One can find in the literature reports about the adhesion of proteins on cellulose ester membranes [3]. However, studies about the formation of cellulose ester ultrathin films have been seldom reported. Cellulose esters are nontoxic materials largely used as coating layers, fibers, inks and films.

Extended abstract of a paper presented at Microscopy and Microanalysis 2013 in Indianapolis, Indiana, USA, August 4 – August 8, 2013.

Multispectral analysis combined with the Periodic Acid-Schiff method was used to investigate cytological features of insoluble polysaccharides and changes in total insoluble polysaccharide content (TPC) during pollen development in rice, including four cytoplasmic male sterility lines (MSLs) and their corresponding fertility-maintaining lines (FMLs). The multispectral curves of the relative transmittance value (RTV) and the images of developing pollen cells were obtained across a range of successive wavelengths (400–720 nm). A minimum RTV was found near 550 nm indicating an absorption peak of the TPC. Thus, the TPC was measured using the RTV of 550 nm. In the four FMLs, the minimum TPC of developing pollen cells occurred at the late microspore stage, while the maximum TPC occurred at the mature pollen grain stage. The TPC levels of pollen cells were significantly higher in the four FMLs than in their corresponding MSLs during and after pollen abortion. Notably, a steep decrease of multispectral curves at 420 nm appeared before the occurrence of abortion, implying a marker associated with pollen abortion in rice. Our results will be helpful for exploring the changes in TPC during pollen ontogenesis in rice and provide a novel method for the study of bio-macromolecules.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Extended abstract of a paper presented at Microscopy and Microanalysis 2012 in Phoenix, Arizona, USA, July 29 – August 2, 2012.

Biological scanning electron microscopy is increasingly performed at low beam energies in order to improve image contrast, reduce charging artifacts, and minimize beam induced damage to the sample. It is natural then to wish to also use these same low accelerating voltage for X-ray microanalysis using an energy dispersive spectrometer (EDS) but a variety of fundamental physical effects affect the performance that can be achieved. An X-ray photon can only be emitted when the incident beam Eo energy exceeds the critical excitation energy Ecrit for that line. As the beam energy is reduced the number of elements that can be excited falls and it is becomes necessary to use L- and M-lines rather than the K-lines accessible at higher energies. At 5keV, the upper limit of ‘low voltage’ microscopy, K-lines can be excited from elements up to calcium, L-lines can be detected up to cesium, and the rest of the periodic table is available using M-lines. The entire periodic table is therefore, in principle, available at an overvoltage U>2, where U = E0/Ecrit But at lower energies the number of accessible excitations falls and some elements cannot, with current technology, then be analyzed and for general purposes an incident energy lower than about 3keV is probably too limiting to be useful.

Terrestrial experiments involving detached growth of GeSi crystals by the Bridgman technique have been performed in preparation for microgravity experiments to be conducted on the International Space Station. These experiments typically produce large GeSi crystals that are sectioned for microscopy, microprobe analysis, and electron-backscatter diffraction analysis. Accurate microanalysis of these crystals is essential for feedback to further growth experiments, but is problematic as the Ge-Si binary exhibits continuous solid solution and it is therefore difficult to prepare homogeneous intermediate standards. There are no experimental measurements on standards of known composition in the Ge-Si binary that could be used to evaluate analytical accuracy. The accuracy of analysis in mineral, oxide, and semiconductor systems has previously been studied using experimental x-ray intensity measurements, correction algorithms and their derived α-factors, and mass absorption coefficient data sets. This study continues that approach and includes results of Monte Carlo calculations in the Ge-Si binary.

TiAl alloys are being developed for future use as lightweight, high strength structural materials for the aerospace and automotive industries. The growth of titanium borides in-situ in a TiAl melt can lead to a variety of structures, such as TiB (Bf and B27 structures) and TiB2, depending on the Ti/Al ratio in the melt, and on the presence of further alloying additions [1,2]. The morphologies of the boride particles can be significantly altered if they grow coupled with other boride or intermetallic phases [2-4]. Since the morphology and distribution of the borides with the intermetallic matrix can significantly affect their effectiveness as creep reinforcements, the structures and growth mechanisms of these composites need to be quantified at the atomic scale.

In a Ti-45.5at.%Al-1.6at.%Fe-l.lat.%V-0.7at.%B alloy know to contain TiB2 precipitates [2,3], the morphology of the boride particles have been investigated using HREM. The Stuttgart JEOL-ARM, operated at 1250kV using the side-entry pole-piece with 0.12nm point resolution, enables the distinction between individual boron and titanium columns in the titanium borides to be achieved for the first time.

Carbon tubes or spheres synthesized by arc-discharge are usually mixed with other byproducts, prohibiting direct measurements of their physical properties by the well established optical techniques because a large quantity of pure specimen is required. Electron energy-loss spectroscopy (EELS) is a unique technique that can be applied to probe the electronic structure of a single carbon tube or sphere. In this paper, the classical dielectric response theory is applied to calculate the EELS spectra acquired from a graphitic carbon sphere at various impact parameters. Graphite is an anisotropic dielectric medium whose dielectric function is described by a tensor. A graphitic carbon sphere is composed of concentric graphitic shells whose dielectric tensor in the spherical geometry, under the local response approximation, is given by (Figure 1)

where are the dielectric function of graphite for an electric field perpendicular and parallel, respectively, to the c axis. In the non-relativistic approximation, the surface excitation is calculated by [1]

Extended abstract of a paper presented at Microscopy and Microanalysis 2008 in Albuquerque, New Mexico, USA, August 3 – August 7, 2008