Biology Commons^™

Apoptosis And Calcification, Kookmin M. Kim

Scanning Microscopy

Calcification in necrosis has long been known. Of the tissue components, the cells are most vulnerable. Nevertheless, little attention has been paid to the role of cell death in calcification. This review attempts to update the mechanism of calcification with an emphasis on the role of apoptosis in calcification. A brief review on the basic sciences relevant to calcification is followed by a discussion of abnormal Ca²⁺ and P_i homeostasis in cell injury and apoptosis. Concomitant increases in Ca²⁺ and P_i in blebs (and matrix vesicles) formed by apoptotic and/or necrotic cells are apparently the primary …

Interactions Of Low-Energy Electrons With Atomic And Molecular Solids, Leon Sanche

Scanning Microscopy

Low energy electrons are involved in a large number of analytical techniques for material analysis either as secondary particles or as the primary excitation source. The interaction of these electrons near the surface of solids can be investigated with high-resolution low-energy electron-beam techniques. The results of experiments performed on atomic and molecular solids in the range 0-30 eV with such techniques are reviewed in the present article. The major types of experiments are briefly described and examples of the results obtained from them are given to illustrate the basic mechanisms which control the electron-solid interactions and to provide a description …

Thermal Stability Of Polystyrene Latex Self-Assembled Arrays Studied By Atomic Force Microscopy, Igor Nevernov, Claudio Nicolini

Scanning Microscopy

Preliminary observation of the heat softening of self-assembled polystyrene latex arrays on the submicron scale using atomic force microscopy is reported. Samples of latex self-assembled arrays have been studied in the temperature range from the room temperature up to 120°C. It has been shown that the submicron scale structure remains stable up to the softening point of polystyrene. The experimental set-up used is described and the results obtained are discussed.

Low Temperature Scanning Electron Microscopy Of Clay And Organic Constituents And Their Relevance To Soil Microstructures, C. Chenu, D. Tessier

Scanning Microscopy

Low temperature scanning electron microscopy (LTSEM) is essential for studying the microstructure of small-sized soil constituents such as clays and organic matter. These hydrated and swelling materials naturally undergo drastic changes in water potential and water content in soils. The cryofixation of clays and organic macromolecules by immersion in cryogens and their LTSEM observation are evaluated. The microstructures of clay minerals, polysaccharide macromolecules and their associations, as revealed by LTSEM, contribute to a better understanding of the water retention properties and physical stabilities of these major soil constituents.

Induction Of Crystallization Of Calcium Oxalate Dihydrate In Micellar Solutions Of Anionic Surfactants, H. Füredi-Milhofer, L. Tunik, N. Filipovic-Vincekovic, D. Skrtic, V. Babic-Ivancic, N. Garti

Scanning Microscopy

Calcium oxalate dihydrate (CaC₂O₄.(2+x)H₂O; COD; x ≤ 0.5) does not readily crystallize from electrolytic solutions but appears as a component in crystalluria. In this paper, we review in vitro studies on the factors responsible for its nucleation and growth with special attention given to the role of surfactants. The following surfactants were tested: dodecyl ammonium chloride (cationic), octaethylene monohexadecylether (non-ionic), sodium dodecyl sulfate (SOS, anionic), dioctyl sulphosuccinate (AOT, anionic), and sodium cholate (NaC, anionic). The cationic and some of the anionic surfactants (SOS, AOT) induced different habit modifications of growing calcium oxalate crystals by …

Work Function Dependence Of Charge Transfer In Desorption And Sputtering Of Atoms From Surfaces, Sarah J. Steuber, Peter Nordlander, Hongxiao Shao, David C. Langreth

Scanning Microscopy

Using a recently developed many-electron theory, we investigate the work function dependence of charge transfer during desorption and sputtering of atoms from metal surfaces. We investigate the effects of substrate bandwidth, atomic velocity and level degeneracy on the charge transfer. We show that many-electron interactions introduce relatively small but measurable effects on the work function dependence of the charge transfer. We find that these effects can be stronger for negative ion states than for positive ion states. The reason is that for negative ions, a strongly correlated Kondo state may be formed near the surface.

Endoarticular Loose Bodies And Calcifications Of The Disk Of The Temporomandibular Joint: Morphological Features And Chemical Composition, C. Piacentini, C. Marchetti, A. Callegari, M. Setti, G. Bernasconi, U. Baciliero, P. Menghini, C. Brusotti

Scanning Microscopy

We studied articular disks and endoarticular loose bodies taken from patients suffering from different types of temporomandibular joint (TMJ) pathology. The scanning electron microscopy (SEM) analysis of the disks and the endoarticular loose bodies was followed by a chemical-compositional analysis using an energy dispersive spectrometer (EDS) and by characterization of the crystalline phases by X-ray powder diffraction (XRD). The articular disks were composed of a central radio-opaque area lacking any evident structural features, surrounded by compact bundles of collagen fibers. EDS and XRD analyses showed that endodiscal radio-opaque areas were hydroxyapatite. By SEM, we observed a fibrous network only in …

Low Temperature Techniques As A Tool In Plant Pathology, Sigrun Hippe-Sanwald

Scanning Microscopy

In plant pathology, low temperature preparation techniques now appear to be feasible methods to stabilize the dynamic ultrastructure of the host-(plant)-pathogen (fungi) interaction for an analysis by transmission electron microscopy. A well defined ultrastructure of small organisms (fungi) and large biological samples such as plant material and as well as the plant-pathogen (fungus) infection sites are presented. The mesophyll tissue of Arabidopsis thaliana is characterized by homogeneously structured cytoplasm closely attached to the cell wall. Infection sites of stem rust (Puccinia graminis f. sp. tritici) on primary leaves of wheat (Triticum aestivum) and powdery mildew (Erysiphe …

Atomic Force Microscopy Of Neuron Networks, A. Cricenti, G. De Stasio, R. Generosi, P. Perfetti, M. T. Ciotti, D. Mercanti

Scanning Microscopy

We imaged uncoated neuron networks by an atomic force microscope in the repulsive regime of contact mode. Images of granule cells and their axons have been clearly revealed with details smaller than 20 nm. The good stability of the sample and the mechanical reproducibility of the microscope allowed the imaging of a neuron culture area of several square microns. By combining tens of images, we were able to reconstruct a highly defined neuronal network. Furthermore, the images were very reproducible over repeated scanning acquisition, demonstrating the mechanical and thermal stability of the instrument-sample system.

Human Skin Physiology Studied By Particle Probe Microanalysis, B. Forslind, M. Lindberg, K. G. Malmqvist, J. Pallon, G. M. Roomans, Y. Werner-Linde

Scanning Microscopy

Particle probe methods (electron probe and proton probe X-ray microanalysis) have been applied to investigate the distribution of elements and water over the different layers of the epidermis. For major elements, electron probe X-ray microanalysis (XRMA) provides the advantage of superior spatial resolution, but for trace element analysis the more sensitive proton probe (particle induced X-ray emission, PIXE) analysis has to be used. On a dry weight basis, the concentration of S is rather constant across the epidermis, whereas the concentrations of P, K, Cl and Na show gradients with high levels in stratum germinativum (basale) and stratum spinosum but …

Comparison Of Dna Fragmentation And Color Thresholding For Objective Quantitation Of Apoptotic Cells, D. R. Plymale, D. S. Ng Tang, C. D. Fermin, D. E. Lewis, D. S. Martin, R. F. Garry

Scanning Microscopy

Apoptosis is a process of cell death characterized by distinctive morphological changes and fragmentation of cellular DNA. Using video imaging and color thresholding techniques, we objectively quantitated the number of cultured CD4 + T-lymphoblastoid cells (HUT78 cells, RH9 subclone) displaying morphological signs of apoptosis before and after exposure to γ-irradiation. The numbers of apoptotic cells measured by objective video imaging techniques were compared to numbers of apoptotic cells measured in the same samples by sensitive apoptotic assays that quantitate DNA fragmentation. DNA fragmentation assays gave consistently higher values compared with the video imaging assays that measured morphological changes associated with …

Imaging Of Cherenkov And Transition Radiation From Thin Films And Particles, N. Yamamoto, A. Toda

Scanning Microscopy

Cherenkov radiation and transition radiation, which are generated by high energy electrons with constant velocity, can be detected in a transmission electron microscope using a cathodoluminescence (CL) detection system. The characteristic peaks due to interference were observed in the emission spectra from thin films of mica, silicon and silver, and their dependence on sample thickness and accelerating voltage was studied. Particles of BaTiO3 and MgO also showed characteristic feature in the spectra which changed with their size. A recently developed imaging system revealed the two-dimensional intensity distribution of these radiations; for example, oscillating contrast, such as equal thickness contour appears …

Crystalline Composition Of Equine Urinary Sabulous Deposits, M. Díaz-Espiñeira, E. Escolar, J. Bellanato, J. A. Medina

Scanning Microscopy

The composition and crystal morphology of 141 equine sabulous deposits were determined by infrared spectroscopy (IR), scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX). The IR analysis revealed that all investigated deposits contained calcium carbonates (calcite, CaCO₃, and/or vaterite, CaCO₃) as major constituents; 42 samples were composed of calcite and vaterite, 33 of calcite, 18 of calcite/vaterite and calcium oxalate, and 17 of vaterite. The remaining specimens contained calcite/vaterite and other compounds (calcium phosphates, sulphate and/or oxalates and/or silica). The examination of 44 selected samples by means of SEM/EDX, revealed the characteristic morphology and …

Photon Emission Induced By The Scanning Tunneling Microscope, Richard Berndt

Scanning Microscopy

By using the tip of a scanning tunneling microscope (STM) as a local source for electrons (or holes) light emission can be excited from metals, semiconductors and molecules. Using this technique, it is possible to combine the high spatial resolution of STM with optical techniques. We review results obtained using a variety of modes of measurements including fluorescence spectroscopy, isochromat spectroscopy and simultaneous mapping of photon emission and surface topography. In spatial maps of the photon emission, clear contrasts are observed with lateral resolutions below 1 nm which are related to the geometric and electronic structure of the sample and …

Cells Are Not Just Bags Of Water: A Personal Appreciation Of The Work Of Dr. Brij L. Gupta And His Contribution To Biological X-Ray Microanalysis, Alice Warley

Scanning Microscopy

This paper surveys the contribution made by the work of Dr. B.L. Gupta to the science of biological X-ray microanalysis. A brief biographical sketch of Brij's early years is given, this is followed by considerations about the models for water transport across epithelia. The ultrastructural and histochemical studies carried out by Brij Gupta and colleagues are reviewed to introduce the historical need for the use of EPXMA for the study of ion and water transport in epithelia. The outstanding contribution made by Brij Gupta's work in this field is outlined, and his understanding of the subcellular distribution ions in other …

The Effect Of Ionizing Irradiation On Type I Collagen Of The Tail In Growing Mice: A Histology And Electron Microscopy Study, M. Q. Yang, E. Kjellén, C. H. Håkansson, M. Palmegren

Scanning Microscopy

In order to examine the effect of radiation on growing tissue, especially the fibroblasts and their end-product, the collagen fibres, tails from 24 mice were irradiated at an age of 8 days with 20 Gy and 30 Gy (>°Co). Tails from 18 animals served as controls. Six mice from each group were sacrificed on day 8, 20 and 30. Transmission electron microscopy was used to examine the fibroblasts and the collagen fibrils. Non-irradiated fibroblasts had a nucleus rich in chromatin and an abundant endoplasmic reticulum with cistemae and condensing vacuoles.

On day 20, approximately 50 % , and on …

Elicitor-Induced Resistance In Tomato Plants Against Fungal Pathogens: Ultrastructure And Cytochemistry Of The Induced Response, Nicole Benhamou

Scanning Microscopy

The introduction of gold cytochemistry in plant pathology has opened new avenues for accurately localizing molecules thought to play important biological functions in the plant metabolism. In an effort to understand the process associated with the induction of plant disease resistance, we investigated the effect of fungal elicitors on the cellular response of tomato root tissues during attack by Fusarium oxysporum f. sp. radicis-lycopersici, the causal agent of tomato crown and root rot. Chitosan was found to induce marked morphological and ultrastructural changes in the fungus grown in pure culture, including cytoplasm aggregation and abnormal deposition of an amorphous …

Morphological Effects Of Glycosaminoglycans On Calcium Oxalate Monohydrate Crystals, Y. Shirane, Y. Kurokawa, Y. Sumiyoshi, S. Kagawa

Scanning Microscopy

The effects of individual glycosaminoglycan (GAG) species on calcium oxalate monohydrate (COM) crystal growth were studied in vitro by the observation of crystal morphology grown in a supersaturated calcium oxalate solution in the presence of GAGs using optical and scanning electron microscopes. GAGs affected the morphology of COM crystals differently depending on the species. The growth rates of the crystals formed in the presence of chondroitin-6-sulfate (ChS-C) were higher in length and lower in width and thickness than those of control crystals. The incorporation of dermatan sulfate or heparin into the crystals formed in the presence of these GAGs was …

Vitamin B6 Deficiency And Galactose Induced Alterations In Morphology And Osmotic Fragility Of Rat Erythrocytes, P. Kaul, H. Sidhu, S. K. Thind, S. K. Sharma, R. Nath

Scanning Microscopy

Recently, red blood cells have been investigated mainly for alterations in ion transporting capacity, membrane bound enzymes or modifications in the structure of its individual constituents in clinical and experimental urolithiasis. However, the implication of such modifications on the physical state or morphology of cells has not been investigated. Scanning electron microscopic studies performed in vitamin B₆ deficient and/or galactose fed rat (established hyperoxaluric models) erythrocytes, showed the presence of large number of stomatocytes, spherocytes and other variously deformed cells as compared to discocytic cells seen in normal control group. These changes in shape were in concurrence with red …

The Formation Of Filopodia-Like Protrusions During Preparation Of Cell Suspensions For Scanning Electron Microscopy, Yuri A. Rovensky

Scanning Microscopy

The standard preparation of cell suspensions (e.g., blood cells, cell suspensions derived from monolayer cultures or from various tissues, etc.) for scanning electron microscopy (SEM) includes fixation of the cells in suspension and subsequent dehydration and critical point drying (CPD) of the cells after their preliminary attachment to the special substrata. In the course of the SEM examination of cell suspensions of various origins, unusual morphological cell surface structures, filopodia-like protrusions (FLP), were consistently detected in 3-20% of the cells in the populations. FLP can be effectively observed only by using a stage tilt angle of no less than 30°. …

Comparative Analysis Of Patch Lesions In The Chick Inner Ear Following Acoustic Trauma: Optical Versus Scanning Electron Microscopy, H. J. Adler, J. Mantooth, Y. Raphael

Scanning Microscopy

Neonatal chicks were exposed to an octave band noise with a center frequency of 1.5 kHz at 116 dB SPL for 4 hours. Seven days following overstimulation, the birds were sacrificed. Their basilar papillae were re-moved, fixed in 4% paraformaldehyde, and processed in two steps. First, the ears were immunostained with a supernatant of mouse anti-tectorial membrane antibodies, followed by a diaminobenzidine process. Examinations of the papillae under an optical stereo microscope revealed a patch site with a partially regenerated tectorial membrane (referred to as the honeycomb).

After the optical studies, the same ears were post-fixed in 1% osmium tetroxide, …

Consistency In Calibrated Backscattered Electron Images Of Calcified Tissues And Minerals Analyzed In Multiple Imaging Sessions, E. G. Vajda, J. G. Skedros, R. D. Bloebaum

Scanning Microscopy

Pure metal standards have been used to calibrate the operating environment in quantitative backscattered electron (BSE) imaging of mineralized tissue, allowing comparisons to be made between various mineralization states of bone at the microscopic level. It has not previously been documented that calibration procedures produce consistent, reliable results over multiple imaging sessions. In this study, BSE images were obtained from bones, pure metals, and a naturally occurring mineral in multiple imaging sessions over a six day period. The graylevel histogram profile (GHP) from each specimen was analyzed for changes in the shape and relative placement on the graylevel spectrum. Computer …

Backscattered Electron Imaging And Energy-Dispersive X-Ray Microanalysis Studies Of Evidence For Calcium Salt Heterogeneity In Fifteen Gallstones From An Elderly Human, Tetsuo Kodaka, Ryoichi Mori, Kazuhiro Debari, Reiji Takiguchi, Shohei Higashi

Scanning Microscopy

We examined 15 variably-sized gallstones, taken from an elderly male, by backscattered electron imaging and energy-dispersive X-ray microanalysis to learn the structural and distribution patterns of gallstone calcium (Ca-) salts. Of the 13 cholesterol-rich stones, nine stones had peripheral concentric layers of Ca-carbonate, whereas 2 stones had peripheral layers of Ca-phosphate. No Ca-salts were detected from 2 cholesterol-rich stones. The 2 stones containing Ca-phosphate had no Ca-salt cores, whereas the stones containing Ca-carbonate were separated into 3 different types: two stones with a Ca-carbonate core, four stones with several Ca-bilirubinate cores of glass-like structure, and 3 stones lacking Ca-salt cores. …

Confocal Laser Scanning Microscopic Studies On Alveolar Bone Remodeling With Orthodontic Tooth Movement And Retention, H. Yagishita, S. Iwatsubo, T. Aoba

Scanning Microscopy

Alveolar bone reconstruction in growing dog during the retention period following orthodontic tooth movement was studied. Three beagle dogs (8-10 kg body weight, about one-year-old) were used and two of the animals were subjected to histological observation. The upper 2nd and lower 3rd premolars on both sides were extracted prior to the orthodontic treatments. After a healing period of one month, the upper 3rd premolar and the lower 4th premolar on the right side were moved mesially with a conventional orthodontic force for 8 weeks, and then retained in their new position for 4 weeks. The contralateral corresponding premolars were …

Analysis Of Glycosaminoglycans Induced In Newly Formed Calcium Oxalate Crystals Using An Undiluted Urine System, Koji Suzuki, Ryuzo Tsugawa

Scanning Microscopy

The aim of this study was to examine the effect of sodium pentosan polysulfate (SPP) in an undiluted urine system and to study its relative affinity to calcium oxalate (CaOx) crystals in the presence or absence of heparan sulfate (HS). CaOx crystals were induced with an overload of oxalate above the metastable limit in spun and filtered urine (SF) and ultrafiltered urine (UF). Then, the crystals were dissolved with EDTA (ethylene-diaminetetraacetic acid), electrodialysed and lyophilized. The polyanions, HS or SPP were added to the UF prior to the addition of oxalate. Polyanions in crystal matrices were examined by cellulose acetate …

Improved Methods For Preserving Macromolecular Structures And Visualizing Them By Fluorescence And Scanning Electron Microscopy, Paul B. Bell Jr., Barbara Safiejko-Mroczka

Scanning Microscopy

To determine the optimal procedures to preserve cytoskeletal and other macromolecular structures for microscopic studies we have evaluated the effects of various methods to extract cultured cells. In this report, we compare results using different fixatives, crosslinking reagents, and permeabilization methods on (1) the labeling of cells for fluorescence microscopy with phalloidin or antibody against tubulin; and (2) the morphological preservation of macromolecular structures for scanning electron microscopy. Maximal labeling of F-actin with phalloidin was obtained by fixing cells in 4% para formaldehyde (PFA) and labeling the unextracted cells with methanolic phalloidin, whereas maximal labeling of tubulin required prefixation with …

Scanning Electron Microscopy And Energy Dispersive Spectroscopy Analysis Of Calciotraumatic Lines In Rat Labial Dentin After Acute Exposure To Strontium Chloride, Hiroyuki Mishima, Toshiro Sakae, Yukishige Kozawa

Scanning Microscopy

Rats were given strontium chloride (SrCli) intraperitoneally at a dose of 500 mg/kg. The upper incisors were removed after injection of strontium. These incisors were studied by scanning electron microscopy - energy dispersive spectroscopy analysis (SEM-EDS) and light microscopy to examine the calciotraumatic lines of strontium in the rat incisor labial dentin.

At 24 hours after injection of strontium, the calciotraumatic response was observed in the predentin using hematoxylin and eosin (H-E) staining. At 5 days, three layers of calciotraurnatic lines were present in the labial dentin using an H-E staining and backscattered electron imaging in the SEM. The external …

Tip-Induced Modifications In Scanning Tunneling Microscopy And Atomic Force Microscopy, K. Cho, J. D. Joannopoulos

Scanning Microscopy

Tip-induced modifications of microscopic processes in scanning tunneling microscopy (STM) and atomic force microscopy (AFM) of the Si(l00) surface are investigated with ab initio total energy pseudopotential calculations. The results of the calculations lead to a new understanding of the microscopic STM measurement process and the micro-mechanical changes (hysteresis and plastic deformation) in the AFM process. In particular, in the latter case, the results predict that the tip can be used to flip dimers on the surface, from one buckled configuration to the other, reversibly, and without inducing damage to either the intrinsic surface or the tip.

The Morphological Effects Of Two Antimicrobial Peptides, Hecate-1 And Melittin, On Escherichia Coli, William G. Henk, William J. Todd, Frederick M. Enright, Pamela S. Mitchell

Scanning Microscopy

The effects of the 26 amino acid, cationic, amphipathic, antibacterial peptide melittin and hecate-1, a 23 amino acid analog of it, on the gram negative bacterium Escherichia coli were investigated using scanning electron microscopy (SEM), transmission electron micros-copy (TEM), and freeze-fracture. Both peptides killed virtually all bacteria at the peptide concentration and cell density used. TEM and SEM revealed aggregates of bacteria entangled with material extruded from the bacterial surfaces. SEM revealed irregular bacterial surfaces with bleb-like projections. TEM and freeze-fracture indicate that the bacterial inner and outer membranes, as well as the peptidoglycan layer between, were extensively damaged. The …

Use Of Colloidal Gold And Neutron Activation In Correlative Microscopic Labeling And Label Quantitation, B. J. Darien, P. A. Sims, K. T. Kruse-Elliott, T. S. Homan, R. J. Cashwell, A. J. Cooley, R. M. Albrecht

Scanning Microscopy

Albumin was conjugated to 16 nm gold particles (Alb-Au16) and infused into anesthetized pigs to determine if plasma, tissue and bronchoalveolar lavage (BAL) fluid concentrations of gold could be quantitated by neutron activation (Au198). Additionally, transmission electron microscopy (TEM) of lung and liver samples was evaluated for sites of gold distribution and morphological changes. The minimum concentration of gold detected by neutron activation ranged between 1.4 and 1.9 ppb (ng/gm of sample). No gold was detected in the plasma of pigs prior to Alb-Au16 infusion, while mean post infusion concentrations were 1.037 ± 0.69 ppm (μg/gm plasma, ±SD). The concentrations …