Biology Commons^™

The Response Of The Tracheal Epithelium To Concomitant Cis-Diamminedichloroplatinum (Ii) And Radiation. An Electron Microscopic Study In Rabbits, C. H. Håkansson, M. Albertsson, M. Palmegren, J. Ranstam

Scanning Microscopy

The ciliated epithelium of the rabbit trachea was irradiated with daily fractions of 2 Gy up to an accumulated dose of 20 Gy (total dose: 2, 6, 10, 16, or 20 Gy). Fifteen to forty-five minutes before the start of each irradiation 0.3 mg Cis-diamminedichloroplatinum (cis-DDP) was given by intraperitoneal injection to each rabbit. Examinations were carried out 1-10 days after each fractionation schedule, when specimens were taken for morphological investigations.

Scanning electron microscope (SEM) examination showed a gradual development of ciliary damage, from blebs on the cilia to swollen tips, broken and bent cilia and finally an epithelial injury …

Timed Acid Bath Effects On Kidney Podocytes Prepared For Scanning Electron Microscopy, B. J. Moore, K. R. Holmes

Scanning Microscopy

In perfusion fixation of dog kidney glomeruli, observation of tear fractured surfaces by scanning electron microscopy reveals the deposition of "debris" on the surface of glomerular visceral epithelium (podocytes) that is not deposited on other kidney tissue surfaces. It is of concern to remove this debris without damage to tissue surfaces. A 5 minute bath in 8N HCI (hydrochloric acid) following postfixation in 1% OsO₄ (osmium tetroxide) removes only some of the debris. A 10 minute acid bath removes this debris without visible damage to podocyte surface structures such as cilia, plicae or blebs. A 20 minute acid bath …

Transplacental Cardiotoxicity Of Cocaine: Atrial Damage Following Treatment In Early Pregnancy, Jacques Gilloteaux, John P. Dalbec

Scanning Microscopy

Using light, transmission (TEM) and scanning (SEM) electron microscopy, cocaine-induced defects were observed in hamster atria. Compared with controls, the treated atria from neonates show endocardial and myocardial damages as the atrial walls thicken. SEM micrographs show intensive blebbing, damage and incomplete coverage of myocardium by the endocardial endothelium. TEM data demonstrate blebs, thinning, and other endothelial cell injuries and complement the SEM findings. Areas of endothelial sloughing may facilitate the formation of luminal and mural thrombi as noticed in many neonatal atria. Adjacent subendocardial myocardial cells display contraction bands, swellings, and vacuolizations. Local and large areas of damaged myocardial …

Copper Toxicity In Erwinia Amylovora : An X-Ray Microanalytical Study, N. Hodson, D. C. Sigee

Scanning Microscopy

The effect of Cu²⁺ ions on the growth and elemental composition of the phytopathogenic bacterium Erwinia amylovora was investigated by in vitro culture in nutrient broth supplemented with CuSO₄.

No detectable inhibition in bacterial growth occurred with medium supplemented at 10^-4M Cu (compared to control medium with no added Cu), but partial inhibition occurred at 10^-3M Cu (limiting toxicity) and complete inhibition at 10^-2M Cu.

Under conditions of limiting toxicity, incubation in Cu²⁺ ions leads to a reduction in the general synthesis of cell materials and the formation of abnormally …

An Introduction To Monte Carlo Simulations, David C. Joy

Scanning Microscopy

The interaction of an electron beam with a solid can be modeled by the so-called Monte Carlo method. This technique produces a stepwise simulation of the electron trajectory by using random numbers to predict scattering angles on the basis of theoretical probability distributions or empirical models. The physical basis of electron scattering in a solid is described and two generic types of Monte Carlo model are then developed together with suggested examples of their application. An IBM PC compatible disc containing these programs is available from the author.

Imaging Of Magnetic Microstructures At Surfaces: The Scanning Electron Microscope With Spin Polarization Analysis, H. P. Oepen, J. Kirschner

Scanning Microscopy

The scanning electron microscope with spin polarization analysis of secondary electrons has been proven to be a powerful tool for studying magnetic microstructures. Secondary electrons created at the surface of a ferromagnet are spin-polarized and contain information about the sample magnetization and its orientation. The combination of a spin polarization analyzer with a scanning electron microscope yields an unique apparatus for probing magnetic properties on a very small lateral scale. The magnetic resolution of < 40 nm is demonstrated. This type of microscope provides high magnetic contrast, while the surface morphology is strongly or even totally suppressed. The capability of studying magnetic properties of semi- infinite samples as well as in ultrathin films is demonstrated with a Fe(lOO) single crystal, video tape, CoCr perpendicular recording medium and ultrathin cobalt films.

Effects Of Low Energy Beta-Irradiation From Tritiated Water On The Morphology Of 3t3 Fibroblasts, Z. Somosy, Tamara Kubasova, J. Kovács, G. J. Köteles

Scanning Microscopy

Cellular alterations of cultured 3T3 cells irradiated with beta-rays from tritiated water were studied by scanning and transmission electron microscopy. We observed decreased negative surface charges, vacuolization of rough endoplasmic reticulum and Golgi-complex, degeneration of mitochondria, increase of lysosomal activity and changes in distribution and amount of microfilaments in the irradiated cells, that parallelled changes in cell shape.

Preservation And Immunogold Localization Of Lipids By Freeze-Substitution And Low Temperature Embedding, Wim Voorhout, Ida Van Genderen, Gerrit Van Meer, Hans Geuze

Scanning Microscopy

The success of postembedding immunocytochemistry depends largely on the preparation methods. The requirements for structural preservation and immunocytochemistry are in some cases contradictory. This is especially the case in the study of lipid-rich structures and the localization of lipid components. Earlier work on freeze-substitution has shown that this method is very promising for the preservation of lipids and the immunocytochemical localization of lipids at the electron microscopical level.

In this study we show that freeze-substitution in combination with low temperature embedding in Lowicryl HM20 has fulfilled this promise. Lamellar bodies in alveolar type II cells contain about 90% lipids and …

Diffusion Of Water In Biological Tissues, H. E. Rorschach, C. Lin, C. F. Hazlewood

Scanning Microscopy

A method is presented for obtaining simple approximate solutions for the problem of self-diffusion in an ordered array of obstacles. Our results are compared with some previous exact and approximate solutions, and we find that our method agrees well with the exact results over a large range of the volume fraction of the obstructions. It is shown that there is an important distinction between measurements of the diffusion coefficient by the capillary flow method and the spin-echo method. The modifications for the spin-echo case are given and applied to recent measurements on the anisotropy of the self-diffusion of water in …

Reliability Of Intracellular Water And Ion Distributions As Measured By X-Ray Microanalysis - A Review, Thomas Von Zglinicki

Scanning Microscopy

X-ray microanalysis can be an important tool to reveal the spatial relationships between polyelectrolytes, ions, and water as they occur within cells and tissues in vivo. To reach this goal, at least two of these three closely interrelated variables should be measured independently. Moreover, the absence of systematic errors should be proven.

The present review discusses the probability of artificial ion and water shifts between intracellular compartments due to the growth of dendritic ice crystals much larger than the cross-sectioned remnants commonly seen in frozen-dried sections. Considering the possible mechanism of ice crystal growth it is concluded that ions and …

Cryopreparation Of Tissue For Clinical Applications Of X-Ray Microanalysis, Godfried M. Roomans

Scanning Microscopy

A number of diseases is associated with changes in ion and/or water distribution at the tissue or cell level, and X-ray microanalysis can be used to investigate the relationship between the disease process and the changes in elemental distribution. For analysis of diffusible elements by X-ray microanalysis, the tissue has to be prepared by cryotechniques. To carry out studies of this kind in a clinical environment poses a number of problems. Some of these problems occur already before the tissue is frozen, others are caused by the practical and ethical limitations that are imposed on the freezing method itself when …

Freezing: Facts And Hypothesis, J. Dubochet, K. Richter, H.-V. Roy, A. W. Mcdowall

Scanning Microscopy

Hexagonal ice crystals formed in frozen biological specimens are large and branched. They can produce severe structural damage by solute segregation but there are also cases where they seem to cause only minor damage. When cooling is more rapid, cubic ice crystals can be formed. These are small and in general, they cause little damage. These observations can be readily explained with the hypothesis that large hexagonal ice crystals can originate from the rewarming induced transformation of a large number of cubic ice crystals. This transformation would take place without significant solute displacement.

The Ultrastructure Of Cryo-Sections And Intact Vitrified Cells - The Effects Of Cryoprotectants And Acceleration Voltage On Beam Induced Bubbling, P. M. Frederik, P. H. H. Bomans, M. C. A. Stuart

Scanning Microscopy

Chemically fixed pancreas was infiltrated with various cryoprotectants to obtain homogeneously vitrified samples upon cooling. The suitability of these samples for cryo-ultramicrotomy was tested. Contrast was hardly detectable initially in thin cryo-sections but increased upon irradiation, irrespective of the cryoprotectant (glycerol, propylene glycol, methanol) used. Contrast and beam damage were analyzed in vitrified thin films from collagen, phospholipid vesicles and various concentrations of glycerol. Glycerol increased the beam sensitivity of both collagen and phospholipid vesicles, but diminished the contrast between matrix and lipid vesicles or collagen fibers. The effects of glycerol as observed in thin films explain some of the …

Aldehyde Fixation Causes Membrane Vesiculation During Platelet Exocytosis: A Freeze-Substitution Study, E. Morgenstern

Scanning Microscopy

Despite a plethora of reports on the ultrastructure of secretory granule release by exocytosis, the release of coagulant activity from stimulated platelets is still being attributed to membrane vesiculation. Membrane vesiculation and the formation of myelin figures have been shown to be artifacts of glutaraldehyde GA fixation. Cells fixed by direct osmium or rapid freezing are free of such structures. Yet there is still doubt that rapid freezing interferes with vesiculation process. This study has addressed this issue by examining: (1) whether freezing and freeze-substitution affects membrane vesiculation, (2) whether paraformaldehyde-fixation also induces the phenomenon, and (3) whether the aldehyde …

High Voltage Electron Microscopy And Low Voltage Scanning Electron Microscopy Of Human Neoplastic Cell Culture, Marek Malecki

Scanning Microscopy

Improved procedures were developed to correlate cell culture data with the images provided by advanced ultrastructural technologies. These procedures were compatible with the two main types of cellular behavior: adherent, spreading (melanomas, rhabdomyosarcomas) and non-adherent in suspension (leukemias).

The ultrastructure and function of spreading neoplastic cells primarily depend on surface properties of the attaching substrates. Therefore, the films used for cultured cell whole-mount ultrastructural analysis must have adherence features identical to those of standard cell culture vessels. Improved procedures were developed to produce the polystyrene films of required qualities. These films allowed processing of cells for electron microscopy including chemical …

Adsorption Staining Of Freeze-Substituted And Low Temperature Embedded Frog Skeletal Muscle With Cesium: A New Method For The Investigation Of Protein-Ion Interactions, L. Edelmann

Scanning Microscopy

A new adsorption staining method for transmission electron microscopy is described by means of which cellular adsorption sites of alkali-metal ions can be visualized in freeze-substituted and low temperature embedded biological material. The main features of this staining method are: 1) the use of Cs⁺ -ions which are known to accumulate in living cells like K⁺ -ions and 2) the removal of the staining solution from thin sections of the embedded material by centrifugal force. It is shown that sections of freeze-substituted and Lowicryl embedded frog skeletal muscle which has not been treated with chemical fixatives can be …

Strategies For Improving The Cytochemical And Immunocytochemical Sensitivity Of Ultrastructurally Well-Preserved, Resin Embedded Biological Tissue For Light And Electron Microscopy, Jan A. Hobot, Geoffrey R. Newman

Scanning Microscopy

Many techniques for processing tissue into resin are available, varying from conventional room temperature to low temperature procedures. The problem is to choose an appropriate method to suit the biological specimen under study. Room temperature approaches with aldehyde and osmium fixation do not give optimal retention of immunoreactivity. Osmium can be removed from sections, but recovery of immunosensitivity is reduced. Osmium post-fixation can be omitted, but heat polymerization of resins causes tissue extraction and loss of immunoreactivity. Alternative techniques rely on the use of milder polymerization methods and avoid osmium. However, while providing an improvement, this alone is not sufficient …

A Cytochemical Scanning Electron Microscopy Study Of Non-Specific Acid Esterase And Acid Phosphatase Activities In Human Peripheral Blood Lymphocytes, A. Campos, E. Fernández-Segura, J. M. García-López

Scanning Microscopy

We analyzed the distribution patterns of nonspecific acid esterase and acid phosphatase activities with cytochemistry-scanning electron microscopy in backscattered and secondary electron imaging modes in isolated normal human peripheral blood lymphocytes . The analysis of non-specific acid esterase activity in the backscattered electron imaging mode showed, in some cells, focal distribution with a well-defined, homogenous deposit. Two patterns of acid phosphatase activity were evident with the backscattered electron imaging mode, i.e., focal and granular. Peripheral blood lymphocytes showing focal activity of both enzymes presented a smooth surface with few scattered microvilli as seen with the secondary electron imaging mode ; …

Conventional And High Resolution Scanning Electron Microscopy Of Biological Sectioned Material, C. Scala, G. Cenacchi, P. Preda, M. Vici, R. P. Apkarian, Gianandrea Pasquinelli

Scanning Microscopy

Intracellular structures of embedded biological tissues (rat kidney, myocardium and small intestine) were observed by conventional-scanning electron microscopy (C-SEM) and high-resolution scanning electron microscopy (HR-SEM) after glass knife sectioning. C-SEM of semi -thin sections of material processed the same way as conventional transmission electron microscopy (TEM) provided strong backscattered electron (BSE)-dependent, two-dimensional secondary electron images (SEI(-)) which precisely integrated and further extended previous light microscopy (LM) observation of the same specimen. In addition, the three-dimensional (3-D) arrangement of intracellular organelles was appreciated using a mixture of acetone-soluble acrylic resin in place of epoxy resin embedding. Since the identification of such …

Phenotypic Subpopulations Of Macrophages And Dendritic Cells In Human Spleen, Patrick J. Buckley

Scanning Microscopy

Using immunohistochemical techniques and a large number of monoclonal antibodies, the presence and distribution of phenotypic subpopulations of macrophages (Mθs) and dendritic cells in human spleen were assessed. The results of this study show that different subsets of Mθs and dendritic cells are present in the spleen and that some of these occupy discrete microanatomic locations . In the red pulp (RP) certain antigens are expressed by different proportions of uniformly distributed Mθs in the cords . On the other hand, some antigens are present on Mθs that form clusters of variable size within the red pulp . These include …

Innervation And Vascular Pharmacodynamics Of The Mouse Spleen, Frank D. Reilly

Scanning Microscopy

Neurohistochemical and in vivo and electron microscopic methods demonstrated α-and ,β-adrenergic receptors and adrenergic innervation in arterioles and "arterial" capillaries of the mouse spleen. Such innervation and receptors in venules and channels within the red pulp were sparse. Cholinergic innervation and receptors were judged to be absent in the microvasculature. Histamine elicited arteriolar dilation which was blocked by metiamide suggesting the presence of H₂ receptors. However , following blockade of H₂ receptors, histamine produced arteriolar constriction. Serotonin elicited only venular constriction. Lactic acid caused arteriolar constriction; bradykinin and prostaglandins (PG) E₂ and PGF_2α triggered arteriolar constriction, …

Ultrastructural Effects Of Ionizing Radiation On Plant Cells, Á. Keresztes, E. Kovács

Scanning Microscopy

Ultrastructural effects of ionizing irradiation were investigated on the specific constituents of plant cells, primarily in fruits, with reference to mushroom cells and comparable data in the literature. In the cell wall the dissolution of the middle lamellae, probably due to radiation damage of pectin, and irregular thickenings were found. In the vacuole the quantity of inclusions changed, or unusual aggregations appeared, presumably in connection with altered phenolic biosynthesis . In chloroplasts the senescence was inhibited, and dedifferentiation occurred into agranal state. In amyloplasts starch hydrolysis was hind ere d. These platidial effects are considered as visible signs of inhibition …

The Role Of Polar Pili In The Adherence Of Pseudomonas Aeruginosa To Injured Canine Tracheal Cells: A Semiquantitative Morphologic Study, D. E. Zoutman, W. C. Hulbert, B. L. Pasloske, A. M. Joffe, K. Volpel, M. K. Trebilcock, W. Paranchych

Scanning Microscopy

Pseudomonas aeruginosa adheres to respiratory epithelial cells in a highly specific fashion. In order to study the role of P. aeruginosa polar pili in the adherence process we conducted a quantitative morphological electron microscopic examination of P. aeruginosa adherence to SO₂ injured canine tracheal cells in vitro. A pilin lacking background strain of P. aeruginosa PAK (BLP2) was constructed using a gene replacement and it in turn was engineered to express either the pilin gene of P. aeruginosa PAO, PAK , or no pilin gene . After 30 minutes incubation of these bacterial strains with injured canine tracheal …

Diapause In The Earthworm, Aporrectodea Longa: Morphological And Quantitative X-Ray Microanalysis Of Cryosectioned Chloragogenous Tissue, A. J. Morgan, C. Winters

Scanning Microscopy

The earthworm, Aporrectodea longa, which experiences an apparent obligatory physiological resting state (“diapause”) in the temperate Summer, was collected monthly from February through to October. Morphological examination of the chloragocytes, and quantitative electron probe X-ray microanalysis of their constituent chloragosome granules prepared by cryoultramicrotomy and air-dried/smearing, were undertaken over this period. Worms entered diapause in May, and emerged from it between August and September. The long-term energy stores in the form of lipid, in addition to the more usual polysaccharide (glycogen) reserves, were accumulated by early diapause; the lipid was gradually consumed during diapause. In addition, the structure and composition …

Scanning Electron Microscopic Study Of Pulp Stones In Human Permanent Teeth, O. Le May, J. C. Kaqueler

Scanning Microscopy

Scanning electron microscopy was used to examine pulp stones which are small calcified formations found in the coronal and/or radicular part of the dental pulp. Pulp stones range considerably in size and shape. Most are round or oval but others can be irregular and may correspond to a reduced duplicate of the pulp chamber anatany. Both free and attached pulp stones were observed. The surface aspect was variable and frequently exhibited large resorption zones. Three characteristic features were observed on fractures a) no characteristic organization where the mineralized mass is compact and hanogeneous, b) a concentric architecture around an initiating …

A Scanning Electron Microscopic Study Of The Morphology Of Scyphidia Physarum Lachmann, 1856 (Ciliophora: Peritrichida), A. Warren

Scanning Microscopy

The morphology of Scyphidia physarum was investigated by scanning electron microscopy (SEM). Both contracted and uncontracted zooids were examined. The pellicular striations encircled the body at mean intervals of 1.1 µm. Pores were randomly distributed over the body surface with a mean density of 21 pores per 100 µm 2. The telotroch band was represented by three closely spaced striations. The adoral ciliature consisted of a single row of cilia in the haplokinety and three rows in the polykinety. The two sets of cilia were separated by a pellicular ridge. At the end of each row of cilia was a …

Crystal-Induced Inflammation: Studies Of The Mechanism Of Crystal-Membrane Interactions, Helen M. Burt, John K. Jackson, Wendy Wu

Scanning Microscopy

Studies of the interactions of monosodium urate monohydrate (MSUM) crystals and calcium Pyrophosphate dihydrate triclinic (CPPD) crystals with biomembranes have been reviewed. Crystalmembrane binding and crystal-induced membranolysis have been studied using human erythrocytes as a model membrane system. The extent of MSUM-membrane binding was determined by incorporating a hydrophobic, fluorescent probe into the membranes, centrifugation to separate free membranes from membranes with bound crystals and quantitation of free membranes by measuring the total fluorescence intensity. The ability of MSUM and CPPD to hemolyse red cells was used as a measure of the membranolytic potential of the crystals. Fluorescence polarization studies …

Changes In Sodium Concentration In Cardiac Myocytes From Diabetic Rats, Alice Warley

Scanning Microscopy

The effects of streptozotocin induced diabetes on rats were studied. The animals showed an increase in blood glucose concentration and a loss of weight from both the body and the heart. Loss of weight from the heart was less severe leading to an increased heart to body weight ratio. Study of element concentrations by X-ray microanalysis showed that there was an increase in intracellular Na concentration in cardiac myocytes from the diabetic animals, but no change in Mg. These results agree with studies which show changes in Na/K ATPase after the onset of diabetes.

X-Ray Microanalytical Studies Of Freshwater Biota: Changes In The Elemental Composition Of Anabaena Spiroides During Blooms Of 1988 And 1989, S. Clay, D. C. Sigee, E. Bellinger

Scanning Microscopy

Samples of Anabaena spiroides were collected from a eutrophic freshwater lake (Rostherne Mere, Cheshire) during the bloom periods of 1988 and 1989, and analysed by SEM electron probe X-ray microanalysis. Vegetative cells, akinetes and heterocysts consistently had clear peaks of Mg, Si, P, S, Cl, Kand Ca.

In comparison to vegetative cells, akinetes had generally higher elemental mass fractions (particularly K and P) while heterocysts had significantly lower levels of Mg, P, and Ca.

For both years, elemental mass fractions in vegetative cells showed significant changes during the course of the bloom, but these were not consistent from one year …

Detectors For Fluorescence Microscopy, Kenneth R. Spring

Scanning Microscopy

The low light levels originating from living cells viewed in the fluorescence microscope place significant limitations on the spatial and temporal resolution which can be achieved . The development of intensified video cameras has enabled the detection , visualization and measurement of these low level signals . The performance characteristics of popular intensified video cameras are compared and guidelines are given for the selection of the appropriate detector for various experimental requirements. Intensified or cooled CCD cameras appear to be the most suitable device for quantitative imaging at low light levels in fluorescence microscopy.