Biology Commons^™

Hydration-Scanning Tunneling Microscopy As A Reliable Method For Imaging Biological Specimens And Hydrophilic Insulators, M. Heim, R. Eschrich, A. Hillebrand, H. F. Knapp, G. Cevc, R. Guckenberger

Scanning Microscopy

The recently discovered high lateral conductivity of molecularly thin adsorbed water films enables investigation of biological specimens, and even of surfaces of hydrophilic insulators by scanning tunneling microscopy (STM). Here we demonstrate the capabilities of this method, which we call hydration-STM (HSTM), with images of various specimens taken in humid atmosphere: We obtained images of a glass coverslip, collagen molecules, tobacco mosaic virus, lipid bilayers and cryosectioned bovine achilles tendon on mica. To elucidate the physical mechanism of this conduction phenomenon we recorded current-voltage curves on hydrated mica. This revealed a basically ohmic behavior of the J-V curves without a …

Early Post-Radiation Changes Of Red Blood Cell Shape In Rats, V. D. Zharskaya, A. B. Chukhlovin

Scanning Microscopy

Scanning electron microscopy (SEM) of red blood cells in whole blood samples from rats was performed following acute γ-irradiation of animals with 0.25 to 1 Gy. Increased incidence of echinocytosis was observed and found to be dose- and time-dependent. At a higher radiation dose (1 Gy), echinocytosis was revealed within 5 minutes after treatment and persisted up to 3 weeks. The data demonstrate the applicability of SEM for detecting minimal radiation-induced lesions of red blood cells.

Formation Of Hydrated Calcium Oxalates In The Presence Of Poly-L-Aspartic Acid, Jeffrey A. Wesson, Elaine Worcester

Scanning Microscopy

The effect of poly-L-aspartic acid (PA) on the crystal structure of calcium oxalate crystals grown after spontaneous nucleation was evaluated as a function of relative supersaturation and calcium:oxalate ratio in a buffered salt solution, with pH and ionic strength in the range of normal human urine. PA was used as a model for naturally occurring acidic urine proteins that have been shown to inhibit nucleation and growth of calcium oxalate crystals. The crystals grown were characterized by optical microscopy and X-ray powder diffraction. It was observed that calcium oxalate monohydrate was the preferred crystalline form in the absence of added …

Role Of Calcium Oxalate Monohydrate Crystal Interactions With Renal Epithelial Cells In The Pathogenesis Of Nephrolithiasis: A Review, John C. Lieske, Mary S. Hammes, F. Gary Toback

Scanning Microscopy

Renal tubular fluid in the distal nephron is supersaturated with calcium and oxalate ions that nucleate to form crystals of calcium oxalate monohydrate (COM), the most common crystal in renal stones. How these nascent crystals are retained in the nephron to form calculi in certain individuals is not known. Recent studies from this laboratory have demonstrated that COM crystals can bind within seconds to the apical surface of renal epithelial cells, suggesting one mechanism whereby crystals could be retained in the tubule. Adherence of crystals to cells along the nephron may be opposed by specific urinary anions such as glycosaminoglycans, …

Crystallization And Stone Formation Inside The Nephron, Dirk J. Kok

Scanning Microscopy

A model is presented visualizing the events leading to calcium-salt, crystal- and stone-formation inside the nephron. For each nephron segment, handling of urine components relevant to stone formation is considered and urine composition determined. This information was applied to nucleation experiments simulating passage of urine through a nephron. The model and in vitro experiments suggest that within normal transit times for the respective nephron segments, particles of a hydroxyapatite-like material first form near the bend in the Loop of Henle of juxtamedullary nephrons. From there on, calcium oxalate particles start to appear: first dihydrate, then monohydrate. In the collecting duct …

Micromorphology Of The Fibers Behind The Frons Plate And Its Adjacent Regions In The Oriental Hornet (Hymenoptera, Vespinae), Jacob S. Ishay, Alina Landsberg, Smadar Pelah

Scanning Microscopy

The present study describes the fibers occurring in the space extending behind the frons plate, or the region between the ocelli and the clypeal plate of the Oriental hornet. These fibers connect to the brain in the anterior part of the head in a zone which is demarcated by an imaginary line traversing the upper part of the two omrnatidia. Static fibers are present in the upper part and in the center of the space. These static fibers do not move with acceleration of the structures attached to the head. They are perpendicular to the frons plate, are relatively numerous …

Development Of An Scanning Tunneling Microscopy-Based Electron Beam Induced Current (Ebic) Microscope, P. Koschinski, V. Dworak, P. E. West, L. J. Balk

Scanning Microscopy

Measurements of electron beam induced currents (EBIC) can either be performed in a scanning electron microscope (SEM) or in a scanning tunneling microscope (STM), since both microscopes are very similar in their basic assembly. However, a straightforward application of an STM in EBIC-measurements, i.e. the use of a microscope tip as a fine source for low energetic electrons is not possible due to the specific demands on the instrument in an EBIC application. The present paper gives a compilation of these demands and describes their conversion into an optimized STM-EBIC microscope.

Numerical Simulation Of The Influence Of Electron Beam Induced Excess Charge Carriers On Potential And Charge Carrier Density Distributions In Gaas Mesfet, K. Kaufmann, L. J. Balk

Scanning Microscopy

Such scanning electron microscopy modes as electron beam induced current (EBIC), cathodoluminescence (CL) and scanning electron acoustic microscopy (SEAM) are widely used for the characterization of semiconductor specimens and devices. These methods are based on the generation of excess charge carriers by an electron beam inside of the investigated specimen. Usually, theoretical models used to explain the physics of these methods presuppose the low injection case.

However, in experimental investigations, the low injection condition cannot always be observed. Thus, for the interpretation of such measurement results more complicated simulation models have to be used. Such simulations can be carried out …

Immunomicroscopy: Resin Techniques And On-Section Labelling With Immunocolloidal Gold Or Immunoperoxidase - Planning A Protocol, Jan A. Hobot, Geoffrey R. Newman

Scanning Microscopy

On-section immunocytochemistry is divided into two parts: (i) processing of biological tissue for section microscopy and (ii) immunolabelling of sections. Many of the more successful microscopical methods employ delicate aldehyde fixation of biological tissue followed by "sympathetic" processing into an acrylic resin. Processing regimens do not have to be complicated. Simple and cost effective room temperature protocols utilising partial dehydration have been devised and they can be as effective as the more complex low temperature techniques in preserving both ultrastructure and antigenic reactivity. The embedded material can be investigated by either light or electron microscopy. Frozen sections can be cut …

Studies On The Role Of Urinary Macromolecules In Urolithiasis: Review Of Methodologies And A Proposal For A Standard Reference Crystallization System, A. L. Rodgers, D. Jappie

Scanning Microscopy

In this study, urine from a calcium oxalate kidney stone former was ultrafiltered (10 kD cut-off). Crystallization was induced in the ultrafiltrate and retentate fractions as well as in a sample of the whole urine. The progress of crystallization was monitored by Coulter Counter and flow cytometry techniques. (The latter has not been used in studies of the role of urinary macro-molecules in urolithiasis). Deposited crystals were examined by scanning electron microscopy. Results indicated that urinary macromolecules in this subject are inhibitors of nucleation and aggregation. These results agree with the findings of some workers but disagree with those of …

The Role Of Defects In Semiconductor Materials And Devices, D. B. Holt

Scanning Microscopy

This literature review leads to the conclusion that recently the basis for an understanding of the electrical and optical properties of structural defects in semiconductors, especially in silicon, has begun to emerge. This is due largely to the ability of scanning electron microscopy (SEM) electron beam induced current (EBIC) and cathodoluminescence (CL) to determine the properties of single, well-defined defects in "state of the art" material. However, there are still major differences concerning the physical models to be used to explain different forms of dislocation EBIC contrast variation with temperature and beam current. Basic ideas in this field are emphasized. …

Multiple Labeling In Electron Microscopy: Its Application In Cardiovascular Research, M. M. H. Marijianowski, P. Teeling, K. P. Dingemans, A. E. Becker

Scanning Microscopy

The heart is a muscular pump kept together by a network of extracellular matrix components. An increase in collagens, as in chronic congestive heart failure (CHF), is thought to have a negative effect on cardiac compliance and, thus, on the clinical condition. Conventional electron microscopy allows for the study of cellular and extracellular components and scanning electron microscopy (SEM) can put these structures in three-dimensional perspective. However, in order to study extracellular matrix components in relation to cells, immunoelectron microscopy is superior. We have used this technique in our studies on heart failure. Heart specimens were fixed in 4% paraformaldehyde …

A Novel Method For Viewing Heavy Metal Stained And Embedded Biological Tissue By Field Emission Scanning Electron Microscopy, R. Geoffrey Richards, Iolo Ap Gwynn

Scanning Microscopy

Backscattered electron (BSE) imaging was used to display heavy metal stained biological structures of various embedded specimens. Samples were fixed, stained and embedded in resin blocks as with preparation for the transmission electron microscope (TEM). Blocks were trimmed to center the specimens in a trapezoidal face of up to 5 mm² and their sides painted with conductive silver paint leaving the face uncovered. Blocks were sputter coated with 6-8 nm of silver, chromium or aluminum, with aluminum providing the best specimen contrast in BSE mode. Samples were examined in a field emission scanning electron microscope operated at a high …

Demonstration Of The Fine Structure Of Stereocilia In The Organ Of Corti Of The Guinea Pig By Field Emission Scanning Electron Microscopy, W. L. Jongebloed, E. A. Dunnebier, F. W. J. Albers, D. Kalicharan

Scanning Microscopy

A combined perfusion- and immersion prefixation with glutaraldehyde followed by a tannic acid/arginine/osmium tetroxide (TAO) treatment of the guinea pig cochlea is described for field-emission gun scanning electron microscopy (FEG-SEM) observation of the fine structure of the stereocilia of the organ of Corti. Conventional osmium tetroxide postfixation methods in combination with a thin conductive coating failed to show the fine structure of the glycocalyx of the epithelial lining in the endolymphatic compartment of the cochlea, in particular, on the stereocilia surface. The antennulae-like glycocalyx covering of the stereocilia surface of the more pronounced rows of outer hair cells has been …

Urinary Stone Proteins: An Update, J. P. Binette, M. B. Binette, M. A. Gawinowicz, N. Kendrick

Scanning Microscopy

The discovery of an organic component in kidney stones dates back to 1684. More than 150 years elapsed before the incrustation of this organic component, which is now called the matrix, was proposed as the mechanism of stone formation. The composition of the matrix remained largely unknown until the development of electron microscopy and the advances in biochemistry combined in the 1950's to usher in the modern era of renal stone matrix investigation. Composed mainly of selectively incorporated proteins generally characterized by high glutamic and aspartic acid content and the frequent occurrence of y-carboxyglutamic acid, the matrix dis-plays a variable …

Neural Transplant Staining With Dii And Vital Imaging By 2-Photon Laser-Scanning Microscopy, Steve M. Potter, Jerome Pine, Scott E. Fraser

Scanning Microscopy

We are developing a multielectrode silicon "neuroprobe" for maintaining a long-term, specific, two-way electrical interface with nervous tissue. Our approach involves trapping a neuron (from an embryonic rat hippocampus) in a small well with a stimulation/ recording electrode at its base. The well is covered with a grillwork through which the neuron's processes are allowed to grow, making synaptic contact with the host tissue, in our case a cultured slice from a rat hippocampus. Each neuroprobe can accommodate 15 neurons, one per well. As a first step in studying neurite outgrowth from the neuroprobe, it was necessary to develop new …

Diffuse Neuroendocrine System: Structural And Functional Effects Of Radiation Injury To Amine Precursor Uptake And Decarboxylation (Apud) Cells, I. M. Kvetnoy, V. V. Yuzhakov, A. O. Molotkov, L. N. Bandurko, R. A. Brodsky, N. D. Yakovleva

Scanning Microscopy

The paper presents a review of the results obtained by the authors on the study of external (gamma) and internal (I-131) radiation effects on the functional morphology and linkage of the diffuse neuroendocrine system (DNES) and amine precursor uptake and decarboxylation (APUD) cells of the stomach and duodenum. The investigations performed enabled us to determine that the morphological changes noted in APUD cells had a dose and time dependency. The present study supports the point of view that the radiation initiates serotonin release from APUD cells, which appears to initiate the mechanism of early postirradiation dysfunctions of the gastrointestinal tract …

Calcium Oxalate Crystallization In Urine Of Healthy Men And Women: A Comparative Study, Niels-P. Buchholz, Dong Sun Kim, Phulwinder K. Grover, Rosemary L. Ryall

Scanning Microscopy

This study aimed to compare calcium oxalate (CaOx) crystallization in undiluted urine from healthy men and women with the object of clarifying the difference in stone incidence between the two sexes. Twenty-four hour urine specimens were collected from 37 men and 28 women. Urinary pH, and concentrations of Ca, oxalate and urate were measured, and indices of crystallization determined by Coulter Counter particle analysis following induction of CaOx crystallization by addition of oxalate. The amount of oxalate required to induce crystallization was significantly (p < 0.01) higher in females than in males, as was the overall particle volume deposited after 90 minutes incubation (p < 0 .006). Scanning electron microscopy revealed larger individual crystals in female urine, and a greater degree of crystal aggregation in male urine, although the average overall size of the precipitated crystal particles did not differ between the two sexes. There were no significant differences between men and women with regard to median pH, or Ca and oxalate concentrations, but the median urate concentrations were slightly, but significantly, higher (p < 0.05) in the women's urines than in the men's. It was concluded that the greater risk of CaOx stones in men is related to an increased propensity to nucleate CaOx crystals per se, rather than to a tendency to form larger crystalline particles.

Specimen Preparation Of The Human Cerebellar Cortex For Scanning Electron Microscopy Using A T-Butyl Alcohol Freeze-Drying Device, Teruyuki Hojo

Scanning Microscopy

A freeze-drying device was applied to t-butyl alcohol substituted nerve cells, fibers and synaptic terminals. Ten percent formalin-fixed human cerebellar cortex specimens were postfixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer solution and were rinsed three times in 5% sucrose solution in the same buffer. After the postfixed specimens were dehydrated using a graded series of ethanols and then transferred into a graded series of t-butyl alcohols (freezing point 25.4°C), the t-butyl alcohol substituted specimens were freeze-dried at 15°C and at high vacuum (5x10^-2 Torr). The freeze-dried specimens were sputter coated with gold. Scanning electron …

Stress Corrosion And Stress Induced Surface Morphology Of Epitaxial Films, Michael Grinfeld

Scanning Microscopy

In addition to several new predictions, the general theory of thermodynamic stability of heterogeneous systems with rearrangement has allowed us to understand the roots of several experimental and theoretical results of the past. One of them is an outstanding paper of Asaro and Tiller on stress corrosion cracking by surface diffusion published two decades ago. We compare results of Asaro and Tiller with conclusions of thermodynamic theory of solids with rearrangement and develop some Asaro-Tiller results in the directions dictated by the needs of thin films technology and experiment. A surface diffusion model in a prestressed elastic solid is studied …

Utilization Of An Electron-Beam Tester For Determining Internal Electric Field Profiles In Micro-Structured Thin-Film Semiconductor Devices, A. Jank, M. Jung, M. Lambert, G. Lichter, H. Schmoranzer

Scanning Microscopy

An electron-beam tester was used to determine the depth profile of the internal potential distribution in an a-Si:H solar cell and hence the internal electric field profile. The a-Si:H solar cells were prepared for the measurements with the electron-beam tester by low-energy chemical plasma beam etching through a plasma-resistant mask structured by electron-beam lithography. In contrast to an earlier work, the solar cells were in situ illuminated and the electric field profiles were determined for new, degraded and in situ annealed solar cells at various cell temperatures. The measuring results demonstrate that the electron-beam testing technique in combination with a …

Comparative Scanning, Transmission And Atomic Force Microscopy Of The Microtubular Cytoskeleton In Fenestrated Liver Endothelial Cells, Filip Braet, Ronald De Zanger, Wouter Kalle, Anton Raap, Hans Tanke, Eddie Wisse

Scanning Microscopy

Endothelial fenestrae control the exchange of fluids, solutes and particles between the sinusoidal lumen and the microvillous surface of the parenchymal cells. Fenestrae have a critical dimension in the order of 150-200 nm, making it necessary to use microscopes with a resolution better than the light microscope. Comparative whole-mount preparations of isolated, purified and cultured rat liver sinusoidal endothelial cells (LEC) were studied by scanning electron microscopy (SEM), transmission electron microscopy (TEM) and atomic force microscopy (AFM). Examination of detergent-extracted LEC by SEM and TEM shows an integral cytoskeleton: sieve plates are delineated by a sieve plate-associated cytoskeleton ring and …

Characterizing The Outlines Of Degraded Fine-Particles By Fractal Dimension, Andrew Hunt, David L. Johnson

Scanning Microscopy

Fractal dimension has been used extensively as a descriptor of the rugged outlines of fine-particles. Potentially, it may be a useful parameter for characterizing the outlines of fine-particles which have been subjected to some form of chemical degradation. Here, fractal dimension values have been computed for the outlines of microscopic lead fine-particles both before and after weak hydrochloric acid dissolution experiments. Values obtained for the post-dissolution rugged profiles were greater than those of the pristine fracture grains which had a Euclidean form. The profiles of the degraded fine-particles could be characterized by a single fractal dimension value, or they exhibited …

Recognition And Phagocytosis Of Apoptotic Cells, Luciana Dini, Maria Teresa Ruzittu, Laura Falasca

Scanning Microscopy

Physiological elimination of unwanted cells within the organism occurs via cell death by apoptosis and phagocytosis of these cells represents a key event in the apoptotic process. Macrophages, which are the dedicated phagocytes, and other occasionally phagocytic cells ingest the apoptotic cells while they are still intact, thus preventing the leakage of potentially harmful materials from the dying cells. Although evidence has been presented that the elimination of apoptotic bodies from the tissue operates by means of specific recognition systems, the molecular mechanisms by which an apoptotic cell is recognized are poorly understood. Recent data indicate that phagocyte recognition of …

Scanning Electron Microscopic Study Of The Postnatal Development Of The Rabbit Cochlea, With An Emphasis On Innervation, Hirofumi Morita, Tomoyuki Hoshino, Kunihiro Mizuta, Satoshi Iwasaki

Scanning Microscopy

The development of nerve fiber arrangements of the organ of Corti was studied in rabbits 1, 3, 5, 7 and 12-days-old using thick sections from celloidin-embedded cochleas which were examined under a scanning electron microscope. The arrangements of nerve fibers varied with developmental age. The tunnel spiral bundle was thick and loosely collected in the immature cochlea. The outer spiral fibers were recognized even in the narrow space of Nuel in the one-day-old cochlea. As Nuel's space is extending, the fibers course along the medial side of Deiters' cells. The arrangement of the outer spiral fibers was irregular and sparse …

Volume Determination Of Human Metaphase Chromosomes By Scanning Force Microscopy, Wolfgang Fritzsche, Eric Henderson

Scanning Microscopy

The scanning force microscope (SFM) yields the topography of the investigated surface. A procedure was developed which starts from this three-dimensional information to estimate the volume of a biological specimen. The volume of spread human metaphase chromosomes was determined in air and rehydrated in aqueous buffer. A difference of the determined volume of a air-dried metaphase chromosome set was found compared to values from electron microscopic investigations, and could be correlated with differences in the hydration state of the chromosomes. SFM-based relative volumes of air-dried chromosomes resembles literature data regarding volume range and distribution. Possible application of SFM-based relative volume …

The Phase Effect In Electronic Stopping: A Survey Of The Contributing Physical Processes, P. Bauer, E. Steinbauer, F. Kastner

Scanning Microscopy

The phase effect in electronic stopping deals with the question whether the energy loss of an ion due to the interaction with electrons depends on the state of aggregation of the target. It is commonly accepted that charge changing collisions of the projectile and changes in the electronic states of the target contribute to the phase effect In addition, the energy loss measurements might possibly be influenced by different impact parameter selection in the two experiments (solid and gas phase). Quantitative results of our calculations show that generally the impact parameter selection inherently present in a transmission experiment is quenched …

Evaluation Of The Surface Changes In Enamel And Dentin Due To Possibility Of Thermal Overheating Induced By Erbium:Yag Laser Radiation, Tat̂Jana Dostálová, Helena Jelíková, Otakar Krejsa, Karel Hamal

Scanning Microscopy

This study investigates the thermal danger of Erbium (Er):YAG laser ablation. Classical preparation techniques have many disadvantages caused by unfavorable changes of temperature, pressure, and by mechanical vibrations. The effect of mechanical vibrations and pressure is eliminated by use of laser ablation technique. The purpose of this study was to analyze the side effects resulting from thermal changes of enamel and dentin in extracted human teeth subjected to pulsed Er:YAG laser radiation using both wet and dry ablation methods.

The micrographs of the sections were checked and measured afterwards. The effect of the investigated laser irradiation on the origin of …

Spatial Dependence Of Electron-Hole Pair Creation In Ion-Solid And Electron-Solid Interactions, F. J. García De Abajo

Scanning Microscopy

The problem of electron excitation induced by interaction of charged particles with solids is investigated on theoretical grounds. The excitation probability is calculated both in homogeneous media and at surfaces. The surface wake potential, needed in the latter, is reviewed. The cases of transmission and aloof geometries are considered separately. Surface plasmons are shown to play a crucial role in the latter. An application to coincidence scanning transmission electron microscopy (STEM) experiments is also discussed. Finally, a spatial representation of the excitation probability is presented.

Inter-Alpha-Inhibitor: A Protein Family Involved In The Inhibition Of Calcium Oxalate Crystallization, Fouad Atmani, Jacques Mizon, Saeed R. Khan

Scanning Microscopy

Inter-α-inhibitor (IαI) is a serine protease inhibitor present in human plasma. It has a molecular weight of about 220 kDa which encompasses 3 chains including two heavy chains and one light chain. The light chain, known as bikunin, is responsible for the antitryptic activity of IαI in the inhibition of various enzymes, such as trypsin and chymotrypsin. Under physiologic or certain pathologic circumstances, several macromolecules related to IαI appear in plasma and urine. However, the physiologic role of IαI remains unclear. As far as urolithiasis is concerned, two urinary macromolecules related to IαI have been isolated and shown to be …