Biology Commons^™

Signal Generation And Contrast Mechanisms In Scanning Electron Acoustic Microscopy, N. Kultscher, L. J. Balk

Scanning Electron Microscopy

In scanning electron acoustic microscopy (SEAM) until now the signal generation is explained mainly by an intermediate production of thermal waves. Though this so-called thermal wave approach has proven to give realistic results for metals, from experimental evidence it seems to fail for other material groups such as ceramics, dielectrics, piezoelectrics and semiconductors. As these material groups are of major technological importance, it is necessary to develop theories which help interpreting those SEAM micrographs obtained for these types of material.

In a comparative manner three different models are discussed in this paper, the well known thermal coupling, the piezoelectric coupling …

Elemental Content Of Anagen Hairs In A Normal Caucasian Population Studies With Proton Induced X-Ray Emission (Pixe), B. Forslind, H. K. Li, K. G. Malmqvist, D. Wiegleb

Scanning Electron Microscopy

The elemental content of anagen hair fibers in a Caucasian population of healthy females and males in the age range 10-69 years was performed to constitute a baseline for further studies of pathological conditions. Proton induced X-ray emission (PIXE) analyses were performed on single hair fibers in triplicates from 103 individuals in order to determine sulfur, zinc, calcium, and chlorine content. The hair fibers were all anagen hairs collected from a site little influenced by genetic and hormonal influences 1.5 cm above the right ear of the probands.

An area 5-8 mm from the follicle bottom was chosen for minimize …

Lipofuscin Accumulation In Cultured Non-Dividing Cells As A Function Of Time And Oxygen Tension, Anders Brunmark, V. Peter Collins, Howard Thaw, Ulf Brunk

Scanning Electron Microscopy

Cultivated human glial cells, kept in a state of density-dependent inhibition of growth, accumulate age-pigment (lipofucsin) within their lysosomal vacuomes with the same characteristics as the corresponding pigment observed in vivo. The rate of formation and accumulation of lipofuscin is greatly accelerated under the conditions of routine cell cultivation in comparison to the in vivo event. Lipofuscin is generally considered to be composed of polymerized products of lipid peroxidation and thus it would be reasonable to suggest that factors which influence lipid peroxidation would also alter the rate of lipofuscin formation. Human glial cells were grown in the presence …

Calcium And Cystic Fibrosis, Godfried M. Roomans

Scanning Electron Microscopy

Cystic fibrosis (CF) is a generally lethal, congenital, genetic disease of unknown etiology. It is likely that a defective regulation of ion and water transport in exocrine glands and possibly also in other epithelial cells has a central role in the pathogenesis of this disease. Calcium has been implicated in the basic defect underlying CF because of findings of abnormally high calcium levels in some secreted fluids and some cells of CF patients.

Using X-ray microanalysis, we have demonstrated elevated calcium concentrations in cultured fibroblasts and in goblet cells of the bronchial epithelium of CF patients. A factor produced by …

Data Acquisition And Processing Techniques For Voltage Contrast Measurements, F. A. Dibianca, D. G. Johnson, C. R. Bagnell, E. I. Cole, W. V. Oxford, R. H. Propst, C. A. Smith

Scanning Electron Microscopy

The effects of several data acquisition techniques on the accuracy of voltage contrast measurements are studied. In particular, the effect of using a voltage reference region directly connected to an external voltage source in performing the image intensity-to-voltage mapping of a node whose voltage is to be determined is examined. This is found to allow improved voltage measurement. The actual reference curves were obtained by least squares fitting the measured intensity-voltage reference data alternately to a quadratic and a cubic function. In addition, various mapping algorithms are considered including ones based alternately on the use of unprocessed, subtracted and normalized …

Quantitative Renal Vascular Casting In Nephrology Research, Vincent H. Gattone Ii, Andrew P. Evan

Scanning Electron Microscopy

The present paper describes the use of a quantitative renal vascular casting method to study the changes associated with kidney disease. Several animal models of hypertension (spontaneously hypertensive rat, SHR, with its normotensive rat the Wistar Kyoto, WKY; Dahl salt sensitive DS - hypertensive, and salt resistant DR -normotensive) were examined at time points when the systemic blood pressure was rising (6 and 12 weeks of age) and following renal denervation (in SHR-WKY rats). The SHR appears to have a smaller caliber afferent arteriole at both 6 and 12 weeks of age. This difference is probably not entirely due to …

A Review Of Modern Characterization Methods For Semiconductor Materials, T. J. Shaffner

Scanning Electron Microscopy

The manufacture of solid state devices in the microelectronics industry involves crystal growth, slice preparation, diffusion and implantation, oxide and metal deposition, patterning and etching, probe testing and packaging of completed devices. This simplified process flow is adopted in an overview to briefly outline some established as well as newly developed techniques of materials characterization. The complexity of semiconductor processing has imposed greater stringency criteria which have resulted in the revitalization of old methods as well as the development of new techniques capable of extreme sensitivity and spatial resolution.

Examples given for trace impurity and dopant detection include neutron activation …

Plant Ultrastructure In The Scanning Electron Microscope, Susan H. Barnes, Stephen Blackmore

Scanning Electron Microscopy

Preparative techniques which have been used to study internal details of plant cells in the scanning electron microscope are reviewed. A number of methods have previously been described which involve selective extraction of materials from freeze-fractured surfaces and can be referred to as freeze-fracture and cytoplasmic maceration. One of these techniques which involves an extended period of cytoplasmic maceration with dilute osmium tetroxide has been applied to the study of Cichorium intybus (chicory) pollen ontogeny. The results obtained, including changes in the numbers of mitochondria and the form of endoplasmic reticulum during the course of development demonstrate the value of …

Surface Morphology Of Unused And Used Hydromerr-Coated Intravenous Catheters, A. Bylock, E. Hultman, B. Gustavsson, L. E. Linder, I. Curelaru

Scanning Electron Microscopy

Hydromer^R-coated polyurethane (Erythroflex)^R catheters, unused, or intravenously inserted for 2-20 days, were studied by scanning electron microscopy (SEM). Both unfixed and fixed (2% glutar-aldehyde in phosphate buffer), and air-or critical-point dried (CPD) specimens were investigated. The catheter segments were sputter-coated with approx. 20 nm gold and studied at an accelerating voltage of 20 kV. The specimens were examined for surface depositions, thickness and structure of the Hydromer^R layers, and occurrence of adhering and embedded bacteria.

The outer Hydromer^R layer showed, in the un-used specimens, scratches and fissures, as well as adhering foreign bodies. In used …

Concerning The Problem Of High Depth Resolution Using Ion Sputtering, G. Blaise

Scanning Electron Microscopy

The inhomogeneity of ion bombardment, the angular dependence of sputtering yields and the crystalline orientation of samples are the three main causes of the degradation of resolution with depth. It is possible to reduce these effects by bombarding at low energy (≈ 1 keV). A low-energy ion-bombardment device is described which has been adapted for use on our sputtered thermal-ion source mass spectrometer.

The Quick-Freezing Of Single Intact Skeletal Muscle Fibers At Known Time Intervals Following Electrical Stimulation, Rashid Nassar, Nancy R. Wallace, Isaiah Taylor, Joachim R. Sommer

Scanning Electron Microscopy

Single intact frog skeletal muscle fibers quick-frozen after known time intervals following electrical stimulation are examined electron microscopically in thin sections, after freeze-substitution, in freeze-fracture/etch preparations, and in cryosections prepared for x-ray microprobe analysis. Techniques are described to perform these operations on a single fiber. Notable morphological differences between conventionally fixed and cryopreserved muscle fibers, and between fibers quick- frozen at different post-stimulation intervals are demonstrated.

Scanning Electron Microscopy Of Human Metaphase Chromosomes, T. D. Allen, E. M. Jack, C. J. Harrison, D. Claugher

Scanning Electron Microscopy

Preparative methods for scanning electron microscopy of chromosomes are dependent on the original source of material. Chromosomes extracted from unfixed metaphase cells via isolation buffers tend to show topography and surface morphology which may have been induced by the choice of isolation buffer itself. Furthermore, this type of preparation often precludes any chromosome identification, as many metaphases have been pooled, and also the chromosomes from these preparations are not suitable for the banding techniques regularly used in clinical cytogenetics. Our own approach has been to use the standard cytogenetic approach, starting with methanol-acetic acid fixed, air dried metaphase spreads, allowing …

Deconvolution In Auger Electron Spectroscopy, B. Chornik, H. E. Bishop, A. Le Moel, C. Le Gressus

Scanning Electron Microscopy

Deconvolution calculations have been applied in Auger Electron Spectroscopy to increase resolution and/or to eliminate loss features. We present: i) A short review of the methodology; ii) Recent results obtained in our laboratory in spectroscopy of Al, Ni, Cu, Ag and Te; iii) A discussion on the conditions for the appearance of artefacts originating either in the calculation or the physical processes (emission anisotropy, distribution of electron path lengths, and intrinsic losses).

Analytical Ultra High Vacuum Scanning Electron Microscopy With Field Emission Gun For Surface Study, P. Morin, P. Abraham, C. Bablet, M. Tholomier

Scanning Electron Microscopy

Surface studies can be carried out with a scanning electron microscope (SEM) having an ultra high vacuum specimen chamber. The main application of this SEM was the micro Auger analysis, but it is also interesting to combine the usual surface study technique with SEM observations. Indeed, these latter give valuable in formation about the topographic, chemical and crystallographic aspects of the surface when the secondary, backscattered and transmission SEM modes are used. The SEM performances are increased by the use of a field emission gun, the high brightness beam of this gun gives new observation possibilities such as the imaging …

Cytoskeletal Changes During Adhesion And Release: A Comparison Of Human And Nonhuman Primate Platelets, J. C. Lewis, M. S. White, T. Prater, K. R. Porter, R. J. Steele

Scanning Electron Microscopy

The organization of cytoskeletal proteins in whole-mount adherent platelets from African green monkeys and normal human volunteers has been studied by SEM, high vacuum electron microscopy (HVEM) and conventional (120 kV) electron microscopy. We describe three distinct organizational zones, the Central Matrix, the Trabecular Zone and the Peripheral Web in spread platelets from both sources. The Central Matrix is an ill-defined superstructure of 80-100 Å filaments of short length which enshrouded the granules, dense bodies, mitochondria and elements of the open-channel and dense-tubular systems. The latter, identified through the use of peroxidase cytochemistry with the whole mounts, is an anastomosing …

Comparison Of Preparative Techniques For Scanning Electron Microscopy Examination Of Soybean Seed Coats In Sectional View, Daniel M. Baker, Harry C, Minor, Merton F. Brown

Scanning Electron Microscopy

Various scanning electron microscopy preparative techniques have been used by researchers to examine sectional views of dry, mature soybean (Glycine~ CL.) Merr.) seed coats. Such previously employed techniques were utilized In our preliminary investigations of seed coat structure, but often yielded unacceptable preservation. Consequently, eight preparative techniques were evaluated In an effort to define conditions required to obtain consistently high quality preservation of soybean seed coats In sectional view. Of the eight procedures tested with the cultivar Williams 82, razor sections and mechanical fractures of dry seed coats yielded the poorest definition of anatomical features. Samples soaked In water prior …

Long Term Effects Of Radiation And Combined Modalities On Mouse Lung, David P. Penney, Paul Van Houtte, Dietmar W. Siemann, Wayne A. Rosenkrans Jr., Philip Rubin, Robert A. Cooper Jr.

Scanning Electron Microscopy

The lung appears to be the major dose-limiting organ in radiation of the thorax. Early responses (<1 week) involve the type II pneumocyte and increased surfactant biosynthesis and secretion. Later changes, which appear to be related to the surfactant response, lead to classical radiation pneumonitis, which is often fatal. Animals which survive radiation pneumonitis develop progressive fibrosis, a late-appearing response, which reduces compliance and available air space, and is usually fatal. This study centers on the fine structural changes in the lungs of LAF₁ mice, 63 weeks following various radiation exposures (5-13 Gy). Doses which are subthreshold in evoking surfactant and pneumonitic responses precipitate fibrosis and atelectasis by 63 weeks, and involve type II pneumocyte sloughing and degeneration. Of the two major deterrents to lung irradiation (pneumonitis and fibrosis), these results suggest that fibrosis always follows pneumonitis, but pneumonitis is not a necessary preliminary step to fibrosis. Bleomycin elicits several morphological alterations characteristic of radiation, and, when combined with the latter, appears to exacerbate radiation effects.

Characterization Of Small Metallic Clusters By Electron Energy Loss Spectroscopy, M. Acheche, C. Colliex, P. Trebbia

Scanning Electron Microscopy

Small spherical tin and gallium clusters (diameters in the range 2 to 80 nm), prepared with a liquid metal ion source, were analysed in a scanning transmission electron microscope (STEM) by electron energy loss spectroscopy (EELS). Both volume and surface plasmon excitations were investigated and their dependence on the cluster size interpreted by classical and quantum mechanical models. A blue shift of the volume plasmon energy with decreasing radius R of the cluster was clearly detected. The full width at half maximum (FWHM) of the peak, related to the damping of the volume plasmon excitation, showed a dependence in 1/R. …

Opportunities For Electron Microscopy In Space Radiation Biology, J. T. Lett

Scanning Electron Microscopy

Densely ionizing, particulate radiations in outer space are likely to cause to mammalian tissues biological damage that is particularly amenable to examination by the techniques of electron microscopy. This situation arises primarily from the fact that once the density of ionization along the particle track exceeds a certain value, small discrete lesions involving many adjacent cells may be caused in organized tissues. Tissue damage produced by ionization densities below the critical value also afford opportunities for electron microscopic evaluation, as is shown by the damage produced in optic and proximate tissues of the New Zealand white rabbit in terrestrial experiments. …

An Organic Reference Specimen: Polyacrylonitrile Film Deposited On Oxidizable Metals By Electro-Polymerization, C. Boiziau, G. Lécayon

Scanning Electron Microscopy

Surface investigations of organic materials require the provision of reference specimens for calibrating experimental equipment.

Because of their specific properties (purity, reproducibility, well-known molecular structure and orderly inter-molecular structure, electronic properties associating conductive and insulating behaviour, quality of the film-to-metal bound), polyacrylonitrile films obtained through electro-polymerization on oxidizable metal cathodes (Fe, Ni, Cu) may be used as a first type of reference sample.

Experimental results, obtained by various surface investigation techniques are presented, and their validity evaluated.

A High Efficiency Cathodoluminescence System And Its Application To Optical Materials, A. D. Trigg

Scanning Electron Microscopy

A high collection efficiency spectroscopic cathodoluminescence (CL) system based on an in-line ellipsoidal mirror has been constructed for use on a Cambridge S250 scanning electron microscope (SEM). It can be fitted to or removed from the SEM in about 30 minutes and requires no significant modification of the instrument. It can be used to obtain total CL images, monochromatic CL images or CL spectra with an ultimate spectral resolution of 1 nm.

The system has been applied to the study of doped synthetic quartz crystals, optical fibres and optical fibre preforms, and to yttrium aluminium garnet, bismuth silicon oxide and …

Fundamental Aspects Of Energetic Particle/Solid Interactions, Nicholas Winograd

Scanning Electron Microscopy

The interaction of keV particles with solids has been characterized by the measurement of the angle and energy distribution of sputtered secondary ions and neutrals. The results are compared to classical dynamics calculations of the ion impact event. Examples using secondary ions are given for clean Ni{001}, Cu{001} reacted with O₂, Ni{001} and Ni{7 9 11) reacted with CO, and Ag{111} reacted with benzene. The neutral Rh atoms desorbed from Rh{001} are characterized by multiphoton resonance ionizaton of these atoms after they have left the surface.

Liquid Metal Sources In Ion Microscopy And Secondary Ion Mass Spectrometry, G. L. R. Mair, T. Mulvey

Scanning Electron Microscopy

This paper reports on state-of-the-art developments in liquid metal sources and some of the finer points of their operating characteristics that are especially relevant to the satisfactory functioning of analytical ion scanning microscopes equipped with secondary ion mass spectrometers. Such effects include unwanted emissions from the source and their exclusion by means of filters and mass separators in the ion-optical column. The design of the ion-optical column is also discussed and some applications of this rapidly advancing form of analytical microscopy are described.

Secondary Ion Mass Spectrometry Of Glasses: Aspects Of Quantification, H. Odelius, A. R. E. Lodding, L. O. Werme, D. E. Clark

Scanning Electron Microscopy

SIMS routines have been developed for the analysis of oxide materials, with applications particularly in element profiling of corrosion layers on glasses after weathering or leaching. The possibilities of quantification and reproducibility have been found critically sensitive to the buildup of charge on the insulating specimens. With control of constant specimen potential, relative sensitivity factors in the positive mass spectrum have been determined for about 20 elements in 10 different alkali-borosilicate glasses. Secondary ion yields were studied as functions of the energy range of ions admitted to the analyzer. At relatively low energies, including the top of the energy distribution, …

The Cathodoluminescence Mode Of The Scanning Electron Microscope: A Powerful Microcharacterization Technique, D. B. Holt, F. M. Saba

Scanning Electron Microscopy

By detecting cathodoluminescence (CL) in a scanning electron microscope (SEM), pan and monochromatic micrographs and CL spectral analyses analogous to x-ray mode point analyses can be obtained. Complete microcharacterization requires alternate examination of both micrographs and spectra. New techniques for near infra-red CL and low-temperatures to produce sharp spectra are of increasing importance. CL emission is due to electron transitions between quantum mechanical states so the radiative defects present can be unambiguously identified at liquid helium temperatures. Strongly luminescent impurities can be detected to below one part in 10⁸. This is 10⁴ times as sensitive as the …

A New Approach For Studying Semithin Sections Of Human Pathological Material: Intermicroscopic Correlation Between Light Microscopy And Scanning Electron Microscopy, G. Pasquinelli, C. Scala, G. P. Borsetti, F. Martegani, R. Laschi

Scanning Electron Microscopy

In order to obtain useful and complete information on the study of pathological material, we observed by scanning electron microscopy (SEM) the same semithin sections observed by light microscopy (LM). For this purpose, the specimen must have, at the same time, chromatic and electron dense characteristics.

We thus developed different specimen preparation methods, subjecting the semithin sections to specific polychromatic staining with high atomic number (Z) elements, to monochromatic staining followed by routine contrasting with uranyl acetate and lead citrate, and to specific cytochemical and immunocytochemical procedures.

The specimens were examined in sequence by LM, by SEM equipped with secondary …

Contribution Of Scanning Electron Microscopy And Associated Analytical Techniques To The Study Of Atherosclerotic Disease, R. Laschi

Scanning Electron Microscopy

Human carotid atheroma has been examined by scanning electron microscopy (SEM) utilizing both secondary and backscattered electron detectors and transmission electron microscopy (TEM) both at conventional and high voltage.

Different cytochemical techniques have been used to map elastic fibers, proteoglycans, calcium, non esterified cholesterol. By immunocytochemistry the distribution of factor VIII related antigen and actin has been studied.

With SEM it was possible to detect aspects of carotid plaques not appreciated when using other conventional techniques. With TEM some modifications of the structural and/or functional features of connective tissue macromolecules have been observed. The occurrence of anomalous collagen has been …

Clinical Applications Of Scanning Electron Microscopy In Gastrointestinal Diseases, F. Bonvicini, G. Zoli, M. C. Maltarello, D. Bianchi, G. Pasquinelli, P. Versura, G. Gasbarrini, R. Laschi

Scanning Electron Microscopy

We considered the role of scanning electron microscopy (SEM) in clinical investigation of different gastrointestinal diseases. The following clinical applications of SEM may be suggested on the basis of our original data and those reported in literature:

in peptic ulcer: assessment of the completeness of healing, by observing the mucosal surface architecture of the scars; identification of mucosal changes, namely enterocytic surface membrane alterations, predictive of recurrence;

in coeliac disease: early assessment of the response to gluten-free diet and follow-up of the patients by staging the process of mucosal repair in cerebriform, intermediate and villous patterns;

in ulcerative …

Scanning Electron Microscopy Of The Human Myocardium, S. Siew

Scanning Electron Microscopy

Owing to the impossibility of fixation, in the human being, by means of perfusion in vivo and the rare availability of fresh comparatively intact hearts resected at transplantation, the possibility was explored of using post-mortem material in instances where the autopsies were performed within 2 hours of demise of the patients. Four cases were examined: 2 normal adult hearts and 2 hearts of hypertensive patients in the stage of decompensation. The hearts were fixed by means of perfusion with 2.5% glutaraldehyde in phosphate buffer, pH 7.4. Contiguous blocks were taken for light microscopy (LM) and scanning electron microscopy (SEM) from …

Characteristic Energy Losses With High Energy Electrons Up To 2.5 Mev, B. Jouffrey, J. Sevely, G. Zanchi, Y. Kihn

Scanning Electron Microscopy

Some aspects of the influence of the energy of the incident electrons in electron energy loss spectroscopy (EELS) are considered. It is shown that this method of analysis used in high voltage electron microscopy, permits one to observe, with a better edge jump ratio than at lower accelerating voltages, the characteristic edges. One important question is to eliminate artefacts in the counting and to record only electrons from the true spectrum. Some recent examples are given. One of them concerns extended energy loss fine structures (EXELFS). It seems high voltage electron microscopy (HVEM) could be very useful in this domain.