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PubMed2004

Imaging and analysis of nanowires.

We used vapor-liquid-solid (VLS) methods to synthesize discrete single-element semiconductor nanowires and multicomposition nanowire heterostructures, and then characterized their structure and composition using high-resolution electron microscopy (HRTEM) and analytical electron microscopy techniques. Imaging nanowires requires the modification of the established HRTEM imaging procedures for bulk material to take into consideration the effects of finite nanowire width and thickness. We show that high-resolution atomic structure images of nanowires less than 6 nm in thickness have lattice "streaking" due to the finite crystal lattice in two dimensions of the nanowire structure. Diffraction pattern analysis of nanowires must also consider the effects of a finite structure producing a large reciprocal space function, and we demonstrate that the classically forbidden 1/3 [422] reflections are present in the [111] zone axis orientation of silicon nanowires due to the finite thickness and lattice plane edge effects that allow incomplete diffracted beam cancellation. If the operating conditions are not carefully considered, we found that HRTEM image delocalization becomes apparent when employing a field emission transmission electron microscope (TEM) to image nanowires and such effects have been shown to produce images of the silicon lattice structure outside of the nanowire itself. We show that pseudo low-dose imaging methods are effective in reducing nanowire structure degradation caused by electron beam irradiation. We also show that scanning TEM (STEM) with energy dispersive X-ray microanalysis (EDS) is critical in the examination of multicomponent nanowire heterostructures.

Computer Simulation↗
PubMed1990

Subcellular distribution of copper in the liver of fetal deer in the last month of gestation.

Fetal deer, in the last month of gestation, accumulate high concentrations of copper in the liver. Livers from fetal deer in late gestation were homogenised and fractionated by continuous sucrose density gradient centrifugation. The distribution of copper closely followed that of DNA; approximately two thirds of the metal was localised to the nuclear fractions with the remaining third in the cytosolic fractions. The fractionation procedure was repeated with digitonin, a lysosomal perturbant: lysosomes were disrupted and the marker enzyme, N-acetyl-beta-glucosaminidase, shifted from the particulate fractions to the cytosolic fractions; the distribution of copper was unaffected. Differential centrifugation of homogenate confirmed that approximately two thirds of the copper was associated with the nuclear fraction. Further confirmation of a nuclear localisation of copper was provided by X-ray microanalysis of purified nuclei.

Animals↗
PubMed1983

Standards for measuring spatial resolution in biological X-ray microanalysis.

Lateral resolution in biological X-ray microanalysis is usually severely limited by the necessity to obtain an adequate number of X-ray counts, which requires the use of a relatively large electron beam spot size. Resolution is further degraded by spreading of the beam within the specimen. Although it is possible to make theoretical calculations of resolution, no straightforward and general method of measuring lateral resolution in biological X-ray microanalysis appears to have been proposed hitherto. In this paper, standards are proposed consisting of distinctive protein-bound elements embedded in araldite, and their use is described.

Egg Proteins↗
PubMed1988

X-ray microanalysis of freeze-dried and frozen-hydrated cryosections.

The elemental composition and the ultrastructure of biological cells were studied by scanning transmission electron microscopy (STEM) combined with energy dispersive X-ray microanalysis. The preparation technique involves cryofixation, cryoultramicrotomy, cryotransfer, and freeze-drying of samples. Freeze-dried cryosections 100-nm thick appeared to be appropriate for measuring the distribution of diffusible elements and water in different compartments of the cells. The lateral analytical resolution was less than 50 nm, depending on ice crystal damage and section thickness. The detection limit was in the range of 10 mmol/kg dry weight for all elements with an atomic number higher than 12; for sodium and magnesium the detection limits were about 30 and 20 mmol/kg dry weight, respectively. The darkfield intensity in STEM is linearly related to the mass thickness. Thus, it becomes possible to measure the water content in intracellular compartments by using the darkfield signal of the dry mass remaining after freeze-drying. By combining the X-ray microanalytical data expressed as dry weight concentrations with the measurements of the water content, physiologically more meaningful wet weight concentrations of elements were determined. In comparison to freeze-dried cryosections frozen-hydrated sections showed poor contrast and were very sensitive against radiation damage, resulting in mass loss. The high electron exposure required for recording X-ray spectra made reproducible microanalysis of ultrathin (about 100-nm thick) frozen-hydrated sections impossible. The mass loss could be reduced by carbon coating; however, the improvement achieved thus far is still insufficient for applications in X-ray microanalysis. Therefore, at present only bulk specimens or at least 1-micron thick sections can be used for X-ray microanalysis of frozen-hydrated biological samples.

Animals↗
PubMed1990

The development and application of ultrastructural research in mycology.

Electron microscopy has contributed a great deal to the field of mycology. Fungal ultrastructure has been, and continues to be, a key research element in the study of spore development and germination, host-pathogen interactions, nuclear behavior, and studies of subcellular organelles and organization linking structure and function. Since the earliest research in transmission electron microscopy in the 1950s, mycologists have kept pace with the developments in all areas of electron microscopy and have used them to great advantage in generating fine structural information on fungi. These recent developments include the use of scanning electron microscopy in the 1960s, X-ray microanalysis, cryopreservation and immunoelectron microscopy in the 1970s and 1980s. All of these techniques will continue to provide mycologists with the means to gain morphological and analytical data at the ultrastructural level.

Cryopreservation↗
PubMed1995

Cutaneous sarcoidosis and foreign bodies. An electron probe roentgenographic microanalytic study.

A 60-year-old woman had extensive and recurrent foreign-body granulomatous inflammation of her forehead. The granulomata contained double refractile crystalline material. Electron probe roentgenographic microanalysis identified silicon, calcium, aluminum, potassium, and sulphur in the crystalline material. Silicon-rich particles are generally considered exogenous. A diagnosis of sarcoidosis was confirmed by the presence of bilateral hilar and mediastinal lymphadenopathy due to biopsy-proven nodal replacement by noninfectious noncaseating granulomata. We conclude that foreign matter may operate as a nidus for the formation of granuloma in sarcoidosis and that the presence of polarizable matter in a sarcoid granuloma does not rule out sarcoidosis.

Aluminum↗
PubMed2002

Fine structure and X-ray microanalysis of a red macrophyte cultured under cadmium stress.

Thalli of the red alga Audouinella saviana were exposed to 600 microM Cd2+ (LC50), 1000 microM Cd2+ and 1500 microM Cd2+ (final concentrations) for 5, 10 and 15 days (each dose) by adding cadmium nitrate to the culture medium. Untreated thalli were set in triplicate as controls for each experiment. Ultrastructural modifications due to cadmium ad/absorption were observed by TEM/SEM electron microscopy. SEM-EDS X-ray microanalysis, definining the accumulation sites, was performed on cryoprepared samples. TEM studies showed striking changes in the plasmalemma of treated algae, which became irregular and convoluted. Electron-transparent exocytic vesicles, possibly related to cell wall polysaccharide synthesis, were observed. The appearance of ribosomes and Golgi bodies, not significantly present in the cytoplasms of untreated cells, suggested enhanced protein and carbohydrate biosynthesis. The cell walls lost their initial grooves and became smooth and thick. More or less electron-dense vesicular systems were formed. Electron-dense sphaeroids occurred in the plasmalemma-cell wall interface, in the cell wall itself and in nearby vesicular membrane systems. Many small vacuoles containing large metal complexes were formed. Complexes were then sequestered into a large vacuole. SEM observations demonstrated that the cell wall and the membrane systems were the most involved in the defense responses. EDS-X-ray microanalysis confirmed the presence of cadmium in these compartments. Chloroplasts, where no Cd2+ signal was detected, were the least affected organelles, showing only a partial disorganization after lengthy exposure to high Cd2+ concentrations.

Air Pollutants↗
PubMed1979

Scanning x-ray microanalysis of root canal sealers.

Seven commercially available root canal sealer powders were analyzed by scanning x-ray microanalysis to identify constituent elements in each. Analysis showed that strongly cytotoxic sealers in vitro contained lead or magnesium, while those materials which exhibited mild or moderate cytotoxicity in vitro contained bismuth, zinc, calcium, or silicone.

Dental Cements↗
PubMed1987

Scanning electron microscopy of human drusen.

Drusen are small, yellowish deposits that form under the retinal pigment epithelium (RPE) with senescence or under certain pathological conditions. The present study examined these structures under the scanning electron microscope. Tissue came from four eyes of 66- and 75-year-old donors who demonstrated widespread drusen of the posterior fundus noted on postmortem examination. Specimens were prepared by either detaching the RPE from Bruch's membrane, or by cryofracturing the tissue for cross-sectional views. Drusen appeared to be composed of irregularly-shaped globular masses and of distinct spherical entities. These particles varied greatly in size, and were situated between the RPE's basement membrane and the outer collagenous zone of Bruch's membrane. Surface views showed drusen components to be embedded in the collagenous zone of Bruch's membrane. Pits corresponding to the sizes of the globular and spherical masses imply that some particles were lost during tissue processing. Fractured cross sections of the irregularly-shaped globular masses revealed a homogeneous, granular matrix with no distinct ultrastructural features, while some of the fractured spherical components demonstrated an internal core. Transmission electron microscopic analysis on the same specimens that were subjected to SEM corroborated these observations. Analytical x-ray microanalysis (Kevex, Foster City, CA) in the SEM revealed major peaks for calcium and phosphorous in the crystalline spherical components, and primarily potassium and chloride in the globular structures.

Aged↗
PubMed2004

Neuropeptides, apoptosis and ion changes in prostate cancer. Methods of study and recent developments.

It has been suggested that neuroendocrine (NE) cells provide paracrine stimuli for the propagation of local carcinoma cells and that NE differentiation is associated with the progression of prostate cancer toward an androgen-independent state. Apoptosis comprises a critical intracellular defense mechanism against tumorigenic growth and is associated with a number of changes in the elemental content of the cell. The neuropeptides bombesin and calcitonin, which inhibit etoposide-induced apoptosis, also inhibit the etoposide-induced elemental changes in prostate carcinoma cells. This important fact strengthens the link between apoptosis and changes in the intracellular elemental content. This protective effect on etoposide-induced apoptosis appears to be quite similar in androgen-dependent and androgen-independent cell lines. This confirms that neuropeptides confer antiapoptotic capabilities on non-neuroendocrine cells in close proximity to neuroendocrine cells. It can therefore be speculated that certain neuroendocrine peptides can increase the survival and further growth of neighboring cells and may thereby contribute to the aggressive clinical course of prostate tumors containing neuroendocrine elements. In addition, this correlation provides an objective basis for the study of neuropeptide target points and may be helpful for alternative therapeutic protocols using neuropeptide inhibitors in the treatment of patients with advanced prostatic carcinoma. The culture techniques described were, thus, designed in order to achieve two important goals. First, the development of an in vitro model that allows an approach to neuroendocrine differentiation in prostate cancer and its role in apoptosis blockage. Second, the method has been designed in order to permit rapid cryofixation of intact cell monolayers for subsequent x-ray microanalysis.

Apoptosis↗
PubMed1997

A possible role of collagen fibrils in the process of calcification observed in the capsule of the pineal gland in aging rats.

The relationship between collagen fibrils and calcified concretions exclusively appearing in the pineal gland of adult/aging rats has been investigated. Deposits of lanthanum, which replace calcium ions are distributed along collagen fibrils with a repeating period of about 70 nm. Calcium has been detected histochemically between collagen bundles surrounding extracellular concretions by means of the pyroantimonate method and by X-ray microanalysis. It is associated with phosphorus. The data presented here suggest that collagen fibrils are involved in the genesis and growth of extracellular concretions located in the connective tissue surrounding the pineal gland of aging rats.

Aging↗