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  • Articles  (2,030)
  • Molecular Sequence Data  (1,179)
  • Life and Medical Sciences  (851)
  • 1990-1994  (2,030)
  • Natural Sciences in General  (2,030)
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  • Articles  (2,030)
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  • 1
    Electronic Resource
    Electronic Resource
    Three-dimensional imaging and image analysis of hippocampal neurons: Confocal and digitally enhanced wide field microscopy (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 269-278 
    Details
    ISSN: 1059-910X
    Keywords: Three-dimensional light microscopy ; Brain slices ; Neurobiology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The microscopy of biological specimens has traditionally been a two-dimensional imaging method for analyzing what are in reality three-dimensional (3-D) objects. This has been a major limitation of the application of one of science's most widely used tools. Nowhere has this limitation been more acute than in neurobiology, which is dominated by the necessity of understanding both large-and small-scale 3-D anatomy. Fortunately, recent advances in optical instrumentation and computational methods have provided the means for retrieving the third dimension, making full 3-D microscopic imaging possible. Optical designs have concentrated on the confocal imaging mode while computational methods have made 3-D imaging possible with wide field microscopes using deconvolution methods. This work presents a brief review of these methods, especially as applied to neurobiology, and data using both approaches. Specimens several hundred micrometers thick can be sampled allowing essentially intact neurons to be imaged. These neurons Image analysis in 3-D is as important as visualization in 3-D. Automated methods of cell counting and analysis by nuclear detection as well as tracing of individual neurons are presented. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290403
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  • 2
    Electronic Resource
    Electronic Resource
    Confocal imaging of dendritic Ca2+ transients in hippocampal brain slices during simultaneous current- and voltage-clamp recording (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 279-289 
    Details
    ISSN: 1059-910X
    Keywords: Fluorescence microscopy ; Ca channels ; Pyramidal neurons ; CA1 region ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Changes in the intracellular Ca2+ concentration ([Ca2+]i) within CA1 hippocampal pyramidal neurons were imaged using confocal laser scanning microscopy in conjunction with Ca2+ -sensitive fluorescent indicators. The imaging was performed in thick hippocampal brain slices while simultaneously measuring or controlling electrical activity with sharp microelectrodes or whole-cell patch-clamp electrodes. The combination of imaging and electrophysiology was essential for interpreting the changes in [Ca2+]i. We compared the increases in [Ca2+]i produced by either of two methods-direct depolarization of the cell via the somatic electrode or high-frequency stimulations of synaptic inputs. The increases in [Ca2+]i in the soma and proximal dendrites caused by both methods were of comparable magnitude and they always decayed within seconds in healthy cells. However, the spatial patterns of distal Ca2+ increases were different. Separate sets of synaptic inputs to the same cell resulted in different spatial patterns of [Ca2+]i transients. We isolated and observed what appeared to be a voltage-independent component of the synaptically mediated [Ca2+]i transients. This work demonstrates that the combination of neurophysiology and simultaneous confocal microscopy is well suited for visualizing and analyzing [Ca2+]i within neurons throughout the CNS and it raises the possibility of routinely relating subcellular [Ca2+]i changes to structural and functional modifications. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290404
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  • 3
    Electronic Resource
    Electronic Resource
    Synaptic morphology of substance P terminals on catecholamine neurons in the commissural subnucleus of the nucleus tractus solitarii in the rat (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 177-183 
    Details
    ISSN: 1059-910X
    Keywords: Sinus afferent pathway ; SP interneurons ; Double immunocytochemistry ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ultrastructure of substance P-containing nerve terminals synapsing on catecholamine neurons in the rat commissural subnucleus of the nucleus tractus solitarii (NTScom) was studied using a double immunocytochemical labeling technique. Although there were numerous tyrosine hydroxylase-immunoreactive (TH-I) somata present, substance P immunoreactive (SP-I) cell bodies were only occasionally found in the NTScom. At the light microscopic level, many SP-I terminals were seen closely associated with TH-I dendrites and somata. At the electron microscopic level, SP-I terminals synapsing on TH-I structures were also readily encountered. SP-I terminals contained small, clear, and predominantly spherical vesicles (32 ± 4 nm diameter), as well as large dense-cored vesicles approximately 100 nm in diameter. Postsynaptic TH-I dendritic profiles of various calibers and somata were encountered. These postsynaptic TH-I structures often showed postsynaptic densities. The morphological features of the SP-TH synapses in the present study, that is, the size of synaptic vesicles and the presence of postsynaptic densities, are quite different from those of central carotid sinus afferent synapses reported in our previous study [Chen et al. (1992), J. Neurocytol., 21:137-147]. Therefore, most of the SP terminals of the SP-TH synapses in the NTScom appear not to originate from the carotid sinus afferents. SP-I second-order neurons of the carotid sinus afferent pathway [Chen et al. (1991), J. Auton. Nerv. Syst., 33:97-98] may be one of the possible sources of such terminals. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290216
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  • 4
    Electronic Resource
    Electronic Resource
    Localizing sites of intradendritic electrophysiological recordings by confocal light microscopy (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 310-318 
    Details
    ISSN: 1059-910X
    Keywords: Hippocampus ; Dendrites ; 3-D imaging ; Pyramidal cell ; Neurophysiology ; Confocal microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Studies were undertaken to develop microscopic methods and imaging procedures that would permit identification of sites of intradendritic microelectrode recordings from pyramidal cells in hippocampal slice preparations. Intradendritic recording were obtained with sharp microelectrodes filled with the dye lucifer yellow. Following a recording session a neuron was iontophoretically injected with the dye and imaged by fluorescence videomicroscopy. Images were stored on videotape for later analysis. They provided a record of the location of the microelectrode recording site. After withdrawal of the microelectrode, slices were processed histologically and imaged a second time with a Bio-Rad 600 confocal attachment on an Olympus BH-2 microscope. Confocal images provided detailed anatomical information in three dimensions. In most instances, a clear identification of the recording site was achieved by comparing video images containing the recording electrode and confocal images.Neurophysiological recordings obtained from proximal and distal apical dendrites were markedly different. Proximal dendritic recordings were similar to those obtained from pyramidal cell soma. However, distal dendrites were not electroresponsive when depolarized by intracellular current injection. The techniques described here, or variations that employ patch electrodes, could provide valuable information that should further an understanding of the properties of dendrites in the central nervous system. © 1994 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290408
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  • 5
    Electronic Resource
    Electronic Resource
    Construction and analysis of a database representing a neural map (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 329-343 
    Details
    ISSN: 1059-910X
    Keywords: Sensory map ; Neural map ; Mechanosensory afferents ; Database ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We describe the development and analysis of a quantitative database representing the global structural and functional organization of an entire sensory map. The database was derived from measurements of anatomical characteristics of a statistical sample of typical mechanosensory afferents in the cricket cercal sensory system. Anatomical characteristics of the neurons were measured quantitatively in three dimensions using a computer reconstruction system. The reconstructions of all neurons were aligned and scaled to a common standard set of dimensions, according to a highly reproducible set of intrinsic fiducial marks. The database therefore preserves accurate information about spatial relationships between the neurons within the ensemble.Algorithms were implemented to allow the integration of electrophysiological data about the stimulus/response characteristics of the reconstructed neurons into the database. The algorithms essentially map a physiological function onto a “field” representing the continuous distribution of synaptic terminals throughout the neural structure. Subsequent analysis allowed quantitative predictions of several important functional characteristics of the sensory map that emerge from its global organization. First, quantitative and testable predictions were made about ensemble response patterns within the map. The predicted patterns are presented as graphical images, similar to images that might be observed with activity-dependent dyes in the real neural system. Second, the synaptic innervation patterns from the sensory afferent map onto the dendrites of a postsynaptic target interneuron were predicted by calculating the overlap between the interneuron's dendrites with the afferent map. By doing so, several aspects of the stimulus/response properties of the interneuron were accurately predicted. © 1994 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070290502
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  • 6
    Electronic Resource
    Electronic Resource
    Hyaluronate distribution in the regenerating retinal pigment epithelium of the rabbit: A study using confocal laser scanning microscopy (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 344-349 
    Details
    ISSN: 1059-910X
    Keywords: Epithelium ; Eye ; Hyaluronate ; Microscopy ; Rabbit ; Regeneration ; Retina ; Sodium iodate ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The distribution of hyaluronate (HA) in regenerating retinal pigment epithelium (RPE) of the rabbit was examined using immunohistochemistry and confocal laser scanning microscopy. The goal was to determine if there is a correlation between differentiation and HA expression, like that seen in developing tissues, where HA accumulates and then disappears as the tissue matures. In normal RPE cells HA is associated mainly with the apical surface. In regenerating RPE (produced by i.v. injection of sodium iodate to damage the epithelium, regeneration arising from spared cells), HA exhibits a patchy distribution among the more immature cells and is especially prominent where they overlap or pile up on each other. Where cells are more mature and form a compact monolayer of cells, HA is expressed mainly on the apical surface, as in normal RPE. The accumulation of HA among the more immature cells in the regenerating epithelial sheet supports the hypothesis that HA influences differentiation by suppressing cell-cell associations until the proper time for their formation. © 1994 Wiley-Liss, Inc.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290503
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  • 7
    Electronic Resource
    Electronic Resource
    Color figure section (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. C1 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070290504
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  • 8
    Electronic Resource
    Electronic Resource
    Masthead (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070200101
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  • 9
    Electronic Resource
    Electronic Resource
    Editorial (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 1-1 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070200102
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  • 10
    Electronic Resource
    Electronic Resource
    A personal computer based implementation of the maximum-likelihood method of analysis of electron microscope autoradiographs (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 73-86 
    Details
    ISSN: 1059-910X
    Keywords: Electron microscopy ; Autoradiography ; Maximum-likelihood estimation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The maximum-likelihood (ML) method for the quantitative analysis of electron-microscopic autoradiographs has been shown to be substantially superior to the conventional crossfire (CF) method. It can generate reliable and accurate tracer concentration estimates with far fewer micrographs and produce valid estimates even at counts low enough to preclude the use of the crossfire method while eliminating the need for special ad hoc treatment of narrow membranous structures as well as the secondary verification of the tracer concentration estimates.Despite these significant advantages, the large computational requirements of the ML method has to date hampered its widespread use. In this paper, we present a new line-integration method that allows us to reduce the computational requirements of the ML method to a point where it becomes feasible to implement it on a small computer system of the type typically available to a laboratory user of EM autoradiography. We present the complete line-integration method for the particular case of EM autoradiography with tritium, and show how it can be adapted to other isotopes.We have constructed a software package that implements the complete maximum-likelihood method on the IBM PC class of machines using our line-integration method. Features of this software package which are of particular importance to the research community are device independence, which makes it usable with a large variety of currently available laboratory equipment, and easy portability of the software and data between different computer systems.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070200108
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  • 11
    Electronic Resource
    Electronic Resource
    Utilization of electron microscopic techniques in the in vitro study of adenohypophysial function and regulation (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 136-151 
    Details
    ISSN: 1059-910X
    Keywords: Pituitary ; Adenomas ; Tissue culture ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Morphologic studies of human adenohypophysial cells using immunocytochemistry and electron microscopy have characterized the hormone-producing cell types of the normal gland and pituitary adenomas. The classifications which have emerged allow more accurate clinicopathologic correlations than ever before, but have also raised new questions concerning cytogenesis, pathogenesis, and structure-function correlations. We report the results of studies which marry the conventional morphologic techniques of light microscopy, immunohistochemistry, electron microscopy, and ultrastructural immunocytology with functional analyses using tissue culture and radioimmunoassay of hormones released into culture media. The hormone secretory activity of nontumorous and adenomatous pituitary cells is correlated with their structural features; their secretory responses to several adenohypophysiotropic factors are compared with morphologic alterations which are characterized at the light and electron microscopic levels by morphometric analysis. These studies have shown that hypothalamic stimulating hormones increase hormone release by their target cells and alter the ultrastructural appearance of the affected cells by increasing organelles involved in hormone synthesis. Inhibitory drugs and adrenal and gonadal steroids are capable of suppressing hormone release by some tumors and also give rise to morphologic changes which correlate with the functional inhibition. Hormone release by clinically nonfunctioning adenomas has been characterized and the behavior of these tumor cells in vitro sheds some light on the reasons for lack of clinical symptomatology. The plurihormonal nature of several nontumorous and adenomatous pituitary cell types has been characterized in vitro. The results of these studies provide the basis for more accurate structure-function correlations which can be used to study the hormonal milieu in vivo, to predict the role of pathogenetic factors in pituitary tumorigenesis, and to assess the therapeutic value of stimulating or inhibiting hormones and drugs.
    Additional Material: 12 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070200203
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  • 12
    Electronic Resource
    Electronic Resource
    Cell lineage in molluscan development (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 75-102 
    Details
    ISSN: 1059-910X
    Keywords: Egg ; Polarity ; Morphogenetic plasm ; Cell communication ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cell lineage specification in molluscs is brought about by two mechanisms: the segregation of morphogenetic plasms and inductive cell interactions. The evidence for the existence of morphogenetic plasms is largely circumstantial, but in one species, Bithynia, such a plasm has been identified in the polar lobe that forms at first cleavage. Inductive cell interactions are thought to be a prerequisite for the development of a large number of tissues and organs. The most extensively studied example is the specification of the mesodermal stem cell in Lymnaea and Patella, which occurs between 5th and 6th cleavage through an interaction between one macromere and a large number of micromeres.Both segregation and induction are tuned to the animal-vegetal polarity of the egg, at least during early development. This polarity probably arises during oogenesis and is manifest in regional differentiations of the surface architecture of the egg, in the distribution of inner membrane particles in the plasma membrane, in membrane fluidity characteristics, in ionic conductance properties of the plasma membrane, etc. All these phenomena have in common that they represent properties of the egg surface, suggesting that the polarity of the egg is somehow imprinted into the plasma membrane and the cortex of the egg during oogenesis. © 1992 Wiley-Liss, Inc.
    Additional Material: 25 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220107
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  • 13
    Electronic Resource
    Electronic Resource
    Role of ultrastructural studies in the analysis of cell lineage in the mammalian pre-implantation embryo (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 103-125 
    Details
    ISSN: 1059-910X
    Keywords: Cell lineage analysis ; Mammalian embryo ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ultrastructural studies have contributed significantly to our understanding of cell lineage differentiation in the mammalian pre-implantation embryo. Such studies have documented, and continue to document, morphological, biochemical, and physiological characteristics of the cell lineages established during the pre-implantation period in eutherian embryos, principally that of the mouse. This review evaluates these contributions and identifies areas of study in which ultrastructural analysis is most likely to have an important role in the future. © 1992 Wiley-Liss, Inc.
    Additional Material: 17 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220108
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  • 14
    Electronic Resource
    Electronic Resource
    Ultrastructural imaging of freeze-fractured plant cells in the scanning electron microscope (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 160-169 
    Details
    ISSN: 1059-910X
    Keywords: Freeze-fracture and cytoplasmic maceration ; Chloroplasts ; Pollen ontogeny ; SEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The application of the freeze-fracture and cytoplasmic maceration technique in ultrastructural studies of plant cells is described. A major advantage of the technique is, that by extracting mobile cytoplasmic components from the freeze-fractured cells, surface relief is introduced and three-dimensional information is obtainable. The details of specimen preparation are described and the results obtained are reviewed. The use of chitosan embedding for very small or fragile specimens is described. © 1992 Wiley-Liss, Inc.
    Additional Material: 21 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220205
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  • 15
    Electronic Resource
    Electronic Resource
    Preservation and visualization of molecular structure in detergent-extracted whole mounts of cultured cells (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 130-150 
    Details
    ISSN: 1059-910X
    Keywords: Electron microscopy ; Scanning electron microscopy ; High resolution ; Cytoskeleton ; Biological specimen preparation ; Cultured cells ; Electrophoresis ; Bifunctional crosslinking reagents ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Today's electron microscopes have a resolution sufficient to resolve supramolecular structures. However, the methods used to prepare biological samples for electron microscopy often limit our ability to achieve the resolution that is theoretically possible. We use whole mounts of detergent-extracted cells grown on Formvar-coated gold grids as a model system to evaluate various steps in the preparation of biological samples for high resolution scanning electron microscopy (SEM)Factors that are important in determining the structure and composition of detergent-extracted cells include the nature of the detergent and the composition of the extraction vehicle. Chelation of calcium is extremely important to stabilize and preserve the cytoskeletal filaments. We have also demonstrated both morphologically and by gel electrophoresis that treatment of cells with bifunctional protein crosslinkers before or during extraction with detergent can significantly enhance the preservation of both proteins and supramolecular structures.The methods used to dry samples are a major determinant of the quality of structural preservation. For cytoskeletons freeze-drying (FD) is superior to critical point-drying (CPD), one reason being that CPD samples have to be dehydrated, thereby causing more shrinkage as compared to FD samples. The high pressures to which samples are exposed during CPD may also cause increased shrinkage, and water contamination during CPD causes severe structural damage. We have obtained the best structural preservation of detergent-extracted and fixed cells by manually plunging them into liquid propane and drying over night in a freeze-drayer.The factor that most limits achievement of high resolution in SEM is the metal coat, which has to be very thin, uniform, and free of grain in order not to hide structures or to create artifactual ones. We have found that sputter-coating with 1-3 nm of tungsten (W) or niobium )Nb( gives extremely fine-grained films as well as satisfactory emission of secondary electrons. These samples can also be examined at high resolution by transmission electron microscopy (TEM) and scanning transmission electron microscopy (STEM). The best preservation and visualization of supramolecular structures have been obtained using cryosputtering, in which the samples are freeze-dried and then sputter-coated within the freeze-dryer while still frozen. © 1992 Wiley-Liss, Inc.
    Additional Material: 16 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220203
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  • 16
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    Comparative study of ion milling techniques in cross-sectional transmission electron microscope specimen preparation (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 199-206 
    Details
    ISSN: 1059-910X
    Keywords: VLSIC ; XTEM ; Semiconductor industry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220209
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  • 17
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    Electronic Resource
    Masthead (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992) 
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    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070220301
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  • 18
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    Preparation of aqueous standards for low temperature x-ray microanalysis (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 207-211 
    Details
    ISSN: 1059-910X
    Keywords: Quantitative low temperature X-ray microanalysis ; Homogeneous dispersion in ice ; Aqueous standards ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A technique, using Nuclepore polycarbonate membrane filters as a containing medium for very small volumes of ionic standard solutions, to produce homogeneous ice standards is described. The standards are suitable for use in a scanning electron microscope. The relationship between elemental X-ray counts and ionic concentration is found to be linear. The method is rapid and simple. Minimum detectable concentrations are given. © 1992 Wiley-Liss, Inc.
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    URL: http://dx.doi.org/10.1002/jemt.1070220210
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  • 19
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    Suspected chemoreceptors in coelenterates and ctenophores (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 265-284 
    Details
    ISSN: 1059-910X
    Keywords: Battery cell ; Cnidocil ; Cnidocyte ; Kinocilium ; Onion-root body ; Nematocyst ; Sensory cell ; Stereocilia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Chemoreceptors in coelenterates and ctenophores have not been identified with certainty. Among prospective chemoreceptive cells are the sensory nerve cells, the cnidocyst bearing cnidocytes, and the epitheliomuscular cells that are likely to be involved in feeding or aggression. Both behaviors are mediated by coordinated chemical and mechanical reception. This is reflected in the close apposition of putative chemo- and mechanoreceptors. Among the structures that have been designated as likely chemo- and/or mechanoreceptors are stereocilia, kinocilia, and/or microvilli which are universally present on all the putative chemoreceptor complexes, while gland cells and mucous secretions are prevalent. Evidence that the actin-containing stereocilia are chemically modulated mechanoreceptors is presented for several forms. © 1992 Wiley-Liss, Inc.
    Additional Material: 21 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070220305
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  • 20
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    Cortical ultrastructure and chemoreception in ciliated protists (ciliophora) (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 225-264 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ciliated protists (ciliates) offer a unique opportunity to explore the relationship between chemoreception and cell structure. Ciliates resemble chemosensory neurons in their responses to stimuli and presence of cilia. Ciliates have highly patterned surfaces that should permit precise localization of chemoreceptors in relation to effector organelles. Furthermore, ciliates are easy to grow and to manipulate genetically; they can also be readily studied biochemically and by electrophysiological techniques. This review contains a comparative description of the ultrastructural features of the ciliate cell surface relevant to chemoreception, examines the structural features of putative chemoreceptive cilia, and provides a summary of the electron microscopic information available so far bearing on chemoreceptive aspects of swimming, feeding, excretion, endocytosis, and sexual responses of ciliates. The electron microscopic identification and localization of specific chemoreceptive macromolecules and organelles at the molecular level have not yet been achieved in ciliates. These await the development of specific probes for chemoreceptor and transduction macromolecules. Nevertheless, the electron microscope has provided a wealth of information about the surface features of clliates where chemoreception is believed to take place. Such morphological information will prove essential to a complete understanding of reception and transduction at the molecular level. In the ciliates, major questions to be answered relate to the apportionment of chemoreceptive functions between the cilia and cell soma, the global distribution of receptors in relation to the anterior-posterior, dorsal-ventral, and left-right axes of the cell, and the relationship of receptors to ultrastructural components of the cell coat, cell membrane, and cytoskeleton. © 1992 Wiley-Liss, Inc.
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    A method for rapidly collecting serial sections for light microscopy (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 22 (1992), S. 306-306 
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    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    URL: http://dx.doi.org/10.1002/jemt.1070220309
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    Morphology of olfactory epithelium in humans and other vertebrates (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 49-61 
    Details
    ISSN: 1059-910X
    Keywords: Olfactory neuron ; Neurogenesis ; Plasticity ; Electron Microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Human olfactory epithelium is similar in organization and cell morphology to that of most vertebrate species. The epithelium has a pseudostratified columnar organization and consists of olfactory neurons, supporting and basal cells. Near the mucosal surface there are also microvillar cells. These cells have neuron-like features and may be chemoreceptors. Human olfactory epithelium is not a uniform sensory sheet. Patches of non-sensory tissue often appear in what was thought to be a purely olfactory region. The significance of these patches has not been determined, but they could reflect exposure to environment agents or changes that occur during the normal aging process.In order to better understand the human olfactory system, further knowledge of the normal structure is necessary. This review addresses the morphology of the human olfactory epithelium and the remarkable plasticity of the vertebrate olfactory system. © 1992 Wiley-Liss, Inc.
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    Olfactory epithelium in young adult and aging rats as seen with high-resolution scanning electron microscopy (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 62-75 
    Details
    ISSN: 1059-910X
    Keywords: Olfactory receptor cell ; Supporting cell ; Ultrastructure ; Lipofuscin granules ; Golgi apparatus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The present study uses mainly scanning electron microscopy to demonstrate the three-dimensional internal cell structures of rat olfactory epithelial cells. The aldehyde-prefixed osmium-DMSO-osmium (AODO) method devised by Tanaka and Mitsushima (1984) was applied to the present study to disclose intracellular structures such as endoplasmic reticulum, mitochondria, Golgi apparatus, and lysosomes. The spatial distribution pattern of these structures in olfactory and supporting cells is discussed, paying special attention to the formation of lipofuscin-like granules present in aged rats. © 1992 Wiley-Liss, Inc.
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    The thickness determination of organic crystals under low dose conditions using electron energy loss spectroscopy (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 166-170 
    Details
    ISSN: 1059-910X
    Keywords: Protein crystals ; Crystal thickness ; Paraffin crystals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In the 3-dimensional (3-D) reconstruction of protein crystals with variable thicknesses the electron images and diffraction patterns can only be merged if the crystal thickness is known. Measurement of the thickness using the ratio of the number of inelastically scattered electrons to the number of electrons in the zero loss peak can be accomplished with parallel electron energy loss spectrometry (PEELS). A theoretical analysis of the accuracy of the technique on paraffin crystals of different thicknesses is presented. Our experimental studies with paraffin crystals show the feasibility of measuring a single layer of 47 Å with good accuracy under low dose and low temperature conditions. A simple experimental apparatus is proposed to obtain thicknesses from small regions of unstained protein crystals prior to collecting the 3-D data sets from the unexposed area of the same crystal.
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    URL: http://dx.doi.org/10.1002/jemt.1070210208
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    Background correction of intensifier camera images in the T.E.M. (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 171-173 
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    ISSN: 1059-910X
    Keywords: Transmission Electron Microscope ; Light Intensifier Camera ; Microscopy ; Background correction ; Image Analysis ; Video Microscopy ; Television Camera ; Frame Store ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Several disadvantages of using intensified television cameras to acquire TEM images can be overcome by using background subtraction with a frame store.
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    Ab-initio primitive cell parameters from single convergent-beam electron diffraction patterns: A converse route to the identification of microcrystals with electrons (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 158-165 
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    ISSN: 1059-910X
    Keywords: Least squares refinement ; X-ray analysis ; Cell parameters ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A new method for the ab initio derivation of Buerger-reduced primitive cell parameters from coordinate measurements of spots on single convergent-beam electron diffraction (CBED) patterns is described, which does not involve trial-and-error. The pattern can be taken along any zone axis, and misorientations of the crystallite by as much as a few degrees are taken into account without loss of accuracy. This derivation of cell parameters by least-squares analysis of the measurements has been automated in a program called NRCBED. Present accuracy is about 1% on lengths and 2° on angles, but could be significantly improved by modelling projector lens aberrations, or by using a microscope without a projector lens. With present technology, it is possible to obtain a CBED pattern and a semi-quantitative energy-dispersive X-ray (EDX) analysis simultaneously from a single microcrystal a few hundred Ångströms across. It becomes therefore possible to identify the material of the crystal on a single CBED pattern: a cell parameter database for known compounds is searched with the primitive cell parameters obtained in the above way, and with a mask describing the EDX results qualitatively. Feasibility is demonstrated on a crystallite of CeO2 500 Ångströms across. With this new approach, trial-and-error should disappear from the solution of other long-standing problems: interpretation of X-ray powder patterns for new compounds in the presence of impurity lines, or in the case of multiple phases should become straightforward.
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    Masthead (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070210301
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    Observation of atomic steps on single crystal surfaces by a commercial scanning electron microscope (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 406-412 
    Details
    ISSN: 1059-910X
    Keywords: REM ; Contrast mechanism ; Imaging technique ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Atomic steps on (111) and (100) crystal surfaces of Pt were observed using a commercial scanning electron microscope (SEM) in secondary electron mode. By comparing the SEM images and those by reflection electron microscopy (REM), the observed contrast was confirmed to be that from atomic steps on crystal surfaces. The contrast mechanism is briefly discussed. One application of this imaging technique is also shown.
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    Observation of double line contrast in surface imaging (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 413-425 
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    ISSN: 1059-910X
    Keywords: REM ; RHEED ; Surface resonance condition ; Contrast splitting ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The double line contrast of a single-atom height step observed in surface imaging for a single crystal in reflection electron microscopy is studied under a variety of experimental conditions. It is suggested that this abnormal contrast is directly associated with the dynamical electron diffraction process. The behavior of the double line contrast is closely related to the order of the Bragg reflected beam, and can be observed mostly under one of the two commonly cited resonance conditions. This phenomenon clearly reveals the differences in the surface imaging for various resonance conditions.
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    Preparation and characterization of MgO surfaces by reflection electron microscopy (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 426-438 
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    ISSN: 1059-910X
    Keywords: Reflection high energy electron diffraction ; Surface topography ; Chemical polishing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have employed several different methods to prepare (100) and (111) surfaces of MgO crystals. (100) surfaces prepared by simple cleaving give good reflection high energy electron diffraction (RHEED) patterns and surfaces with a high density of coarse steps. Chemical polishing of this surface results in a roughening of the topography whilst annealing in oxygen considerably smoothens the surfaces although they appear to be contaminated. Under certain conditions we find that the MgO crystals will cleave along the (111) plane. Both cleaved and mechanically polished (111) surfaces are atomically flat and reconstructed after oxygen annealing.
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    Ciliated and microvillar receptor cells degenerate and then differentiate in the olfactory epithelium of rainbow trout following olfactory nerve section (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 22-27 
    Details
    ISSN: 1059-910X
    Keywords: Plasticity ; Retrograde degeneration ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We used scanning (SEM) and transmission (TEM) electron microscopy to examine ultrastructural changes in the olfactory epithelium (OE) of rainbow trout following unilateral olfactory nerve section. Both ciliated receptor cells (CRC) and microvillar receptor cells (MRC) degenerated and subsequently differentiated from unidentified precursor cells. The following changes took place in fish that were held at 10°C at the stated period following olfactory nerve section: on day 7, MRC and CRC contained intracellular vacuoles; on day 12, the olfactory knobs appeared disrupted; by day 26, olfactory receptor cells were absent from the OE; on day 42, there were receptor cell bodies and a few CRC with short cilia at the apical surface; and opn day 55, a small number of both CRC and MRC had differentiated. By day 76, both CRC and MRC repopulated the OE. Degenerative changes in the cytoplasm of the sustentacular cells (SC) and ciliated nonsensory cells (CNC) were observed in the first 26 days following olfactory nerve section, but these cells remained intact throughout the experiment. The degeneration and subsequent differentiation of CRC and MRC supports and extends previous observations that both cells types are olfactory receptor neurons with axons that extend along the olfactory nerve to the olfactory bulb. © 1992 Wiley-Liss, Inc.
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    Ultrastructural neurobiology of the olfactory mucosa of the brown trout, Salmo trutta (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 28-48 
    Details
    ISSN: 1059-910X
    Keywords: Nose ; Olfaction ; Ultrastructure ; Toxicology ; Smell ; Sensory ; Fish ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes four investigations of the olfactory mucosa of the brown trout: 1) the ultrastructure of the olfactory mucosa as revealed by scanning (SEM), conventional transmission (TEM), and high voltage (HVEM) electron microscopy; 2) light and electron-microscopic investigations of retrograde transport of the tracer macromolecule horseradish peroxidase (HRP) when applied to the cut olfactory nerve; 3) SEM and TEM investigations of the effects of olfactory nerve transection on cell populations within the olfactory epithelium; and 4) ultrastructural investigations of reversible degeneration of olfactery receptors caused by elevated copper concentrations. The trout lofactory epithelium contains five cell types: ciliated epithelial cells, ciliated olfactory receptor cells, microvillar olfactory receptor cells, supporting cells, and basal cells. The ciliated and microvillar olfactory receptor cells and a small number of basal cells are backfilled by HRP when the tracer is applied to the cut olfactory nerve. When the olfactory nerve is cut, both ciliated and microvillar olfactory receptor cells degenerate within 2 days and are morphologically intact again within 8 days. When wild trout are taken from their native stream and placed in tanks with elevated copper concentrations, ciliated and microvillar cells degenerate. Replacement of these trout into their stream of origin is followed by morphologic restoration of both types of olfactory receptor cells. Ciliated and microvillar receptor cells are primary sensory bipolar neurons whose dendrites make contact with the environment; their axons travel directly to the brain. Consequently, substances can be transported directly from the environment into the brain via these “naked neurons.” Since fish cannot escape from the water in which they swim, and since that water may occasionally contain brain-toxic substances, the ability to close off - and later reopen - this anatomic gateway to the brain would confer a tremendous selective advantage upon animals that evolved the “brain-sparing” capacity to do so. Consequently, the unique regenerative powers of vertebrate olfactory receptor neurons may have their evolutionary origin in fishes. © 1992 Wiley-Liss, Inc.
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    Use of ferrofluids to obtain magnetic domain images in afm (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 98-99 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    URL: http://dx.doi.org/10.1002/jemt.1070230109
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    Elimination of artifactual labeling of hippocampal mossy fibers seen follong pre-embedding immunogold-silver technique by pretreatment with zinc chelator (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 100-101 
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    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Additional Material: 1 Ill.
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    Flat embedding and immunolabelling of SW 1116 colon carcinoma cells in LR white: An improved technique in light and electron microscopy (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 1-9 
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    ISSN: 1059-910X
    Keywords: Mucin ; Immunofluorescence ; Immunoelectron microscopy ; Monoclonal antibody 19-9 ; Matrigel ; PAS stain ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Human SW 1116 colon carcinoma cells were grown on matrix-covered coverslips and flat embedded in specially prepared gelatin capsules in the hydrophylic resin LR White. Dehydration and polymerization were carried out so as to maximize preservation of antigenicity. Sections were cut perpendicular to the substratum. To visualize mucin, semithin sections of SW 1116 cells were stained with periodic acid Schiff (PAS) reagent for light microscopy, and ultrathin sections were labelled with a monoclonal mucin antibody (Mab 19-9) and immunogold for electron microscopy. Immunofluorescence was carried out on whole cultured cells using Mab 19-9. The morphological preservation of SW 1116 cells embedded in LR White was comparable to that of Epon-embedded cells. Mucin was localized on the microvillar surface of the apical plasma membrane and occasionally in intercellular spaces between adjacent cells. Mucin was also present in vesicles in the apical and lateral part, and to a lesser extent in the basal part of the cells. We conclude that this new technology significantly improves the morphological preservation of cells and tissues in LR White, while also serving to sustain the antigenicity of cellular antigens.
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    Scanning electron microscopy of negatively stained catalase on a silicon wafer (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 32-38 
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    ISSN: 1059-910X
    Keywords: High-resolution scanning electron microscope (HRSEM) ; Negative staining ; Surface ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A high-resolution scanning electron microscope capable of 7 Å spatial resolution at 30-kV accelerating voltage was used to observe negatively stained protein molecules. Thin platelet crystals, densely packed monolayers, and low-density deposits of beef liver catalase were prepared on the surface of silicon wafers and negatively stained with phosphotungstic acid. The tetrameric structure of the catalase molecule was observed for the first time by scanning electron microscopy on the surface of the smooth silicon wafer.
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    Sequential observations followed by acid etching on the enamel surfaces of human teeth under scanning electron microscopy at low vacuum (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 429-436 
    Details
    ISSN: 1059-910X
    Keywords: Low vacuum specimen chamber ; No dehydration ; No coatings ; Backscattered electrons ; Enamel structures ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A scanning electron microscope equipped with a low vacuum specimen chamber and a Robinson's backscattered electron detector was employed to observe the natural surfaces of human buccal enamel before and after 30 percent phosphoric acid etching sequentially up to 90 sec at the same sites with no coatings. Furthermore, successive etching patterns were compared between deciduous and permanent teeth. On the imbrication lines of young permanent teeth, prismend pits surrounded with a “prismless” structure occasionally disappeared after acid etching and became a prismless enamel. Sequential etching caused the prismless areas and the areas of a type 1 etching pattern to decrease, and a cone-shaped prism structure and a complex type of the type 1 and type 2 etching pattern (type 1-2) to appear. The former was a transitional type between the prismless enamel and type 2 prisms. These etched surfaces show type 2 prisms after deeper etching. Small dome-shaped structures, slightly elevated on the attrited enamel surfaces, were found only in deciduous teeth. After acid etching, such areas which retained the prismless enamel rose to the underlying surfaces of cone-shaped prisms. © 1993 Wiley-Liss, Inc.
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    Preservation of antigenic reactivity after cryofixation, cryosubstitution, and cryoembedding in Lowicryl HM23: Application to alcohol dehydrogenase in Drosophila fat body (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 453-454 
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    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Additional Material: 1 Ill.
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    Masthead (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070240601
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    Application of cryotechniques in cartilage tissue preservation an immunoelectron microscopy: Potentials and problems (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 457-464 
    Details
    ISSN: 1059-910X
    Keywords: Cartilage ; Proteoglycans ; Collagen ; Structure ; Freeze substitution ; Immunohistochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cryotechnical processing of cartilage has the potential to solve many of the tissuespecific problems associated with various routine chemical fixation protocols. This is particularly the case with respect to extracellular matrix architecture, the distortion or destruction of which (caused by extraction and/or precipitation of proteoglycan molecules) may be prevented. Adoption of such techniques also permits high-sensitivity immunoelectron-microscopy of the extracellular matrix space (carbohydrate epitopes). However, a number of difficulties still remain to be resolved, particularly that of matrix-cell interface separation occurring during freeze substitution and low temperature embedding. These problems are briefly addressed and possible solutions outlined. © 1993 Wiley-Liss, Inc.
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    Improved preservation of ultrastructure in difficult-to-fix organisms by high pressure freezing and freeze substitution: I. Drosophila melanogaster and Strongylocentrotus purpuratus embryos (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 465-473 
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    ISSN: 1059-910X
    Keywords: High pressure freezing ; Freeze substitution ; Drosophila ; Sea urchin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this study, we have applied the techniques of high pressure freezing and freeze substitution to embryonic cell types which are usually difficult to fix properly for electron microscopy. In both Drosophila and Strongylocentrotus purpuratus, we see improved preservation of both membrane systems and cytoskeleton when compared to published results on the same cells using conventional electron microscope (EM) fixation methods. Finally, we have seen that postembedding labelling of sections is possible even after light osmium fixation during freeze substitution. © 1993 Wiley-Liss, Inc.
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    Freeze substitution after fast-freeze fixation in preparation for immunocytochemistry (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 474-487 
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    ISSN: 1059-910X
    Keywords: Immunogold labeling ; Antigenic preservation ; Bioluminescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: As compared to classical chemical fixation, the physical immobilization of ultrastructures by fast-freeze fixation (FFF) and the subsequent exchange of water in its solid state by freeze substitution (FS) improve the preparation procedure for immunogold labeling (IGL).FFF-FS results in a morphological preservation of unchallenged quality, as well as in a better preservation of antigenic reactivity, thus allowing remarkable precision of labeling on sections.However, FFF, particularly over a cooled metal plate, requires a heavy and expensive machine. It is not suitable for all biological specimens and in the best conditions, which remain difficult to standardize, the thickness of the well-preserved portion of the specimen does not exceed a few μm for compact tissues, and exceptionally 30-40 μm for isolated cells.The FS procedure is long and must be adjusted empirically for every new specimen and antigenic detection. The preservation of a given antigen's reactivity in the presence of fixative agents and embedding resins remains unpredictable. The action of fixative agents is different and milder in FS than when they are used classically in chemical fixation. By chance, one of the best FS procedures for the preservation of both ultrastructure and antigenicity appears to be by using acetone alone, together with a molecular sieve to improve the water exchange process. A large choice of embedding resins usually allows us to find a compromise between ultrastructural and antigenic preservation. © 1993 Wiley-Liss, Inc.
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    Freeze-Substitution for morphological and immunocytochemical studies in insects (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 488-504 
    Details
    ISSN: 1059-910X
    Keywords: Cryofixation ; Cryosubstitution ; Olfactory sensilla ; Thermo-/hygroreceptive sensilla ; Pheromone-binding protein ; Bombyx mori ; Antheraea polyphemus ; Poecilocampa populi ; Boreus hiemalis ; Drosophila melanogaster ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Methods of plunge freezing and freeze-substitution (FS) for insect antennae and similar body appendages are described. In these more or less cylindrical specimens, usually a layer below the cuticular surface of 10-15 μm thickness is well preserved without freezing damage, further inwards ice-crystal ghosts of increasing size are encountered, but in the very centre of antennal branches (diameter ∼80 μm) of the silkmoth, Bombyx mori, freezing damage is usually reduced again. The frost-hardy species, Poecilocampa populi and Boreus hiemalis, exhibit regions free from freezing damage up to 40 μm below the cuticular surface. Secondary freezing damage in silkmoth sensory hairs is observed only after deliberately warming the specimens to -43°C for 〉〉10 min before FS. Secondary artefacts due to the substitution process are investigated by comparison with freeze-etching and by comparing different FS media and protocols. Methanol is not recommended as a substitution medium for insect specimens. Structures particularly liable to substitution damage are the stimulus-conducting pore tubules of olfactory sensilla and the receptor cell membrane. Extraction of soluble components is more likely with pure organic solvents without added chemical fixing agents and with prolonged substitution at elevated temperatures. Such extraction may also be a possible artefact with soluble antigens in immunocytochemical studies. A review is given of the major achievements attained with these techniques in insect functional morphology and immunocytochemistry. © 1993 Wiley-Liss, Inc.
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    Discrepancies in kinematic calculations of HOLZ lines (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 509-513 
    Details
    ISSN: 1059-910X
    Keywords: Convergent-beam diffraction ; Lattice parameter ; Computer simulation ; Electron wavelength ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have found significant differences between the results of computer simulations of HOLZ line patterns. The computations in question are made in the kinematical approximation. After trivial errors are eliminated the programs fall into two groups. There is a discrepancy between the two that increases with distance from the zone axis. The difference is small but not negligible at the level of precision used in determining lattice parameters or strain.We show which of the two is correct in the kinematic approximation and that the discrepancy between the two groups is of the order of the error introduced by dynamical interaction. © 1993 Wiley-Liss, Inc.
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    Morphometric evaluation of populations of neuronal profiles (cell bodies, dendrites, and nerve terminals) in the central nervous system (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 315-337 
    Details
    ISSN: 1059-910X
    Keywords: Computer-assisted image analysis ; Morphometric methods ; Cell clusters ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Morphometric techniques have been developed to quantitatively characterize groups of transmitter-identified neuronal profiles, such as cell groups, dendrite and nerve terminal fields. These morphometric techniques will be illustrated by introducing some general tools for image analysis which can be considered as a background for the present specific applications. The following methods have been included: (1) methods to identify and quantitatively characterize, from both numerical and geometrical standpoints, groups of profiles in a two- and three-dimen-sional frame; (2) methods to evaluate the evenness of a certain distribution of profiles in the plane; (3) methods to identify subgroups of profiles based on their different spatial or optical density; and (4) methods to compare the distributions of two or more groups of profiles. The applications of these general tools to some neuroanatomical problems, such as cell group definition and description, have been illustrated. Practical examples performed on immunocytochemical preparations of neuronal profile populations are also given. Finally, the potentiality of numerical classification to classify and compare morphometric data has been shown. As an example, numerical classification methods have been applied to the morphometric and microdensitometric analysis of adrenaline/neuropeptide Y costoring neuronal systems of the brainstem in adult and aged rats. © 1992 Wiley-Liss, Inc.
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    Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0) (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 338-346 
    Details
    ISSN: 1059-910X
    Keywords: Tutorial ; Electron microscopy ; Light microscopy ; Software ; Quantitative morphology ; Stereology ; Morphometry ; Simulations ; Terminology ; Data types ; Sampling ; Hierarchies ; Interpretation of data ; Bio-Matrix Project ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes a computer-aided tutorial for biological stereology. Stereology, a type of quantitative morphology, includes a collection of statistical methods that quantify the structural compartments that can be viewed in sections with light and electron microscopy. These methods provide volume, surface, length, shape, and number data, and help define the quantitative relationships among the structural compartments of biological hierarchies. Hierarchies, which connect structural data ranging in size from molecules to organs, serve as a central core to which the data of biological databases can be linked. The tutorial focuses on two objectives. It provides the user primarily interested in using quantitative morphology databases with background information, and offers a set of state-of-the-art tools to researchers wishing to use these methods in the laboratory. The main topics of the tutorial include: introduction to quantitative morphology, symbols/terms, data types, sampling, hierarchies, data interpretation, and utilities. The tutorial runs under the MS-DOS operating system and requires at least an IBM PC AT (or compatible), a color monitor (EGA, VGA), 540 KB of RAM, and 3 MB of hard disk space. © 1992 Wiley-Liss, Inc.
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    A new procedure for handling impervious biological specimens (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 355-360 
    Details
    ISSN: 1059-910X
    Keywords: Primer method, Microwave fixation, Seeds, Mites, Whiteflies ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A new application of techniques for preparing impervious biological specimens for light microscopy (LM) and transmission electron microscopy (TEM) has been developed. Microwave irradiation was used to facilitate fixation. A priming technique was used to increase the bonding of the outer surface of the specimens with the resin. Priming the waxy or cuticular surface with Z-6040 (gamma-glycidoxypropyl trimethoxysilane) solved the problem of specimen “pull out” from the resin. Insect specimens with various types of cuticles (waxy or chitinous) and seeds were successfully studied ultrastructurally using this technique. © 1992 Wiley-Liss, Inc.
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    Computer assisted data collection for stereology: Rationale and description of point counting stereology (PCS) software (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 347-354 
    Details
    ISSN: 1059-910X
    Keywords: Quantitative morphology ; Morphometry ; Light microscopy ; Electron microscopy ; PCS System III ; MS-DOS ; UNIX ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The paper describes microcomputer software for point counting stereology. Stereology includes a collection of statistical methods that quantify the images of light and transmission electron microscopy. The methods use test grids placed over images to collect raw data, which includes counts of points, intersections, transections, and profiles. In turn, the counts are included in stereological equations that give estimates of compartmental volumes, surfaces, lengths, or numbers. These parameters describe the composition of a structure in three-dimensional space. The PCS (point counting stereology) System Software III serves as a data collection, storage, and management tool. Users set up point counting protocols without programming, enter data by pressing predefined function (MS-DOS) or alphabetic keys (UNIX), store data in files, select files for analysis, and calculate results as stereological densities. The latest version of the PCS software includes a new user interface and is designed as a research “front end” that can feed data either into the calculation tools of a stereology tutorial (Bolender, 1992, this issue) or into the analysis routines of quantitative morphology databases (Bolender and Bluhm, 1992). © 1992 Wiley-Liss, Inc.
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    Handbook of crystallography for electron microscopists and other, by A. G. Jackson. Springer-Verlag, New York, 1991, 226 pp, $49.50 (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 368-368 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070210413
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    The influence of lens chromatic aberration on electron energy-loss spectroscopy quantitative measurements (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 361-367 
    Details
    ISSN: 1059-910X
    Keywords: Elemental analysis ; Analytical electron microscopy ; Transmission electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: An investigation has been made into the effect of chromatic aberrations of a pre-spectrometer lens system on quantitative elemental analysis by electron energy loss spectroscopy (EELS). In transmission electron microscopy (TEM) diffraction mode, the measured effects are typically 150-330 times larger than if only objectiv-lens chromatic aberration were important. We discuss several methods of avoiding errors arising from chromatic aberration, including selection of a suitable optical mode (dependent on the desired spatial resolution), adjustment of the TEM imaging system so as to focus the system for a chosen energy loss, and analysis of a large area of a uniform specimen. © 1992 Wiley-Liss, Inc.
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    Ultrastructural evidence for the axonal localization of caudodorsal cell hormone mRNA in the central nervous system of the mollusc Lymnaea stagnalis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 12-18 
    Details
    ISSN: 1059-910X
    Keywords: In situ hybridization ; Digoxigenin ; Electron microscopy ; Cryosections ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The technique of in situ hybridization has been used to evaluate the expression of an ovulation hormone mRNA (caudodorsal cell hormone; CDCH) in the central nervous system (CNS) of the mollusc Lymnaea stagnalis. Hybridization with radioactive as well as with nonradioactive labeled oligonucleotide and plasmid probes revealed a specific labeling on cell bodies of caudodorsal cells (CDCs), which are known to produce CDCH, on the light microscopical level. In addition, specific labeling was observed outside the cell bodies, as far as the cerebral commissure, where CDCH is released in the haemolymph. To investigate whether these signals represent an axonal localization of the CDCH mRNA, we performed in situ hybridization at the electron microscopical (EM) level. The results showed an intraaxonal localization of CDCH mRNA with digoxigenin labeled oligonucleotide and plasmid probes. Gold labeling was observed in secretion granules, and double labeling experiments showed that these granules also contain CDCH. This specific intragranular localization suggest that CDCH mRNA is transported through the axon and released by exocytosis in the haemolymph. © 1993 Wiley-Liss, Inc.
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    mRNA structure, in situ, as assessed by microscopic techniques (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 19-28 
    Details
    ISSN: 1059-910X
    Keywords: In situ transcription ; IST ; Translational control ; Stem-loop sequence ; RNA-binding proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The secondary and tertiary structure of RNA, in situ, is thought to be involved in distinct functions such as directing association of the RNA with the cytoskeleton, enzymatic activity of some RNAs, and the control of translation. In situ transcription (IST), a procedure by which cDNA is synthesized in situ, has been used to assess mRNA structure in situ using fixed cells or tissues. Distinct banding patterns were noted for mouse and rat POMC. Unique IST banding patterns were observed when an oligonucleotide complementary to a putative POMC stem-loop structure was used to prime IST. Indeed local changes in banding patterns could be elicited by pharmacological agents which modulate POMC translation. Inhibition of POMC synthesis with NaF or dexamethasone decreased the number of POMC mRNAs in the polysome fractions and increased the intensity of high molecular weight IST-derived bands. Forskolin, a stimulator of POMC synthesis, had the opposite effect. One mechanism by which translational control is thought to occur is by regulation of ribosome movement down the mRNA by specific binding of cytosolic proteins to RNA structure. Cytosolic protein fractions from AtT20 pituitary cells have been shown to specifically bind to the IST-predicted RNA structure. These findings suggest that 1) mRNA structure can be assessed in situ, 2) translation may be altered by the secondary and tertiary structure of mRNAs, and 3) a predicted stem-loop structure exists in situ in the 5′-end of POMC mRNA. © 1993 Wiley-Liss, Inc.
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    Protocol of electron microscope in situ nucleic acid hybridization for the exclusive detection of double-stranded DNA sequences in cells containing large amounts of homologous single-stranded DNA and RNA sequences: Application to adenovirus type 5 infected Hela cells (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 2-11 
    Details
    ISSN: 1059-910X
    Keywords: Double-stranded viral DNA ; Electron microscopy ; HeLa cell ; In situ hybridization ; Lytic infection ; S1 nuclease ; Replication ; Viral Ad5 genomes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In order to gain a further insight into the relationships of the complex process of replication of adenovirus genomes to the substructures which occur in the nuclei of adenovirus type 5 (Ad5) infected HeLa cells, we have visualized directly, at the electron microscopic level, viral double-stranded DNA (dsDNA) in late infected nuclei by the use of a post-embedding in situ hybridization technique with a biotinylated specific DNA probe. The procedure is based on the removal of single-stranded (ss) nucleic acids by S1 nuclease. The highest levels of signal density for viral dsDNA were detected over the fibrils of the large, centrally located viral genome storage site and over the viral nucleoids of both clustered and isolated viruses. Lower but significant signals were observed over the fibrillo-granular network of the peripheral replicative zones, where both transcription and replication of viral DNA occur. On the other hand, the labeling of the enclosed viral ssDNA accumulation sites, also involved in viral replication but not transcription, was negligible, which suggests that, in the latter, the newly synthesized viral dsDNA immediately extends into the adjacent peripheral replicative zone to be transcribed and/or replicated.
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    Quantitative aspects of synapses on Golgi-impregnated neurons (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 334-352 
    Details
    ISSN: 1059-910X
    Keywords: Identified neurons ; Quantification ; Rotating/tilting ; Synaptic contacts ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: With the classical Golgi techniques, numerous types of neurons can be distinguished in the cerebral cortex, each with a specific dendritic geometry and pattern of axonal ramifications. In the present review we describe two techniques which allow quantification of synapses on identified neurons: (1) Golgi-rapid impregnation-gold toning-electron microscopy, and (2) Golgi-Kopsch impregnation-gold toning-electron microscopy in combination with staining of the tissue with ethanolic phosphotungstic acid (E-PTA). Both techniques were applied on neurons in the visual cortex of young and adult rabbits. By means of rotating and tilting specimens in the electron microscope, the nondistinctive ultrastructure of obliquely sectioned synapses can be circumvented, leading to precise estimates of asymmetrical vs. symmetrical synapses without complete reconstruction of the neuron. © 1992 Wiley-Liss, Inc.
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    Intracellular injection in fixed slices in combination with neuroanatomical tracing techniques and electron microscopy to determine multisynaptic pathways in the brain (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 15-30 
    Details
    ISSN: 1059-910X
    Keywords: Lucifer Yellow ; Photoconversion ; Retrograde tracing ; Anterograde degeneration ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Intracellular Lucifer Yellow filling in fixed tissue has been recently introduced as a novel neuroanatomical approach to reveal the detailed morphology of individual neurons in isolated preparations of the central nervous system. Since dye injections are performed under visual control, the method is characterized by a high degree of inherent staining selectivity, thus circumventing the element of randomness often considered to be the crux of classical Golgi-impregnation techniques. Moreover, the opportunity to optically monitor the injection procedure renders fixed slice preparations highly advantageous to be used in combination with retrograde fluorescent tracing. Subsequently, dye-filled neurons may be subjected to a simple photoconversion procedure leading to the intracellular formation of a stable polymer thus obtaining permanent specimens for light microscopy purposes. Due to the osmiophilic nature of the precipitate the photoconverted material is equally suitable for correlated electron microscopy, thus enabling the analysis of neuronal microcircuitry. At the ultrastructural level, sources of afferent input to identified projection neurons may be revealed by lesion-induced anterograde degeneration of synaptic terminals, therefore enabling the direct demonstration of multisynaptic links. Finally, morphologically identified neurons may be immunocytochemically characterized at the pre- and postembedding levels. It is therefore suggested that their methodological versatility and relative technical ease render intracellular fixed-slice injections a promising complement to the catalogue of anatomical techniques. © 1993 Wiley-Liss, Inc.
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    Photoconversion of diaminobenzidine with different fluorescent neuronal markers into a light and electron microscopic dense reaction product (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 2-14 
    Details
    ISSN: 1059-910X
    Keywords: Lucifer Yellow ; DiI ; Fluorescent neuronal tracers ; Retrograde transport ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This article describes methods for photoconverting diaminobenzidine (DAB) into a stable, light and electron microscopically visible dark reaction product in neurons which contain a fluorescent dye. Photoconversion of DAB has been achieved so far with the following fluorescent dyes: rhodamine labeled latex microspheres (RLM), 4,6-diamidino-2-phenylindole (DAPI), 5,7-di-hydroxytryptamine (5,7-DHT), Fast Blue (FB), Nuclear Yellow (NY), Diamidino Yellow (DY), Evans Blue (EB), acridine orange (AO), ethidium bromide (EBR),1,1′-dioctadecyl-3,3,3′,3′-tetramethylindolcarbocyanine perchlorate, D-282 (DiI), propidium iodide (PI), and intracellularly injected Lucifer Yellow (LY). The dye is introduced into the neurons by tinctorial staining, retrograde transport, or intracellular injection. Photoconversion is conducted by incubating the tissue with the fluorescent substance-containing cells in a DAB solution under simultaneous strong illumination with ultraviolet (UV) light. During the formation of the reaction product, the fluorescence disappears from the cell. In all cases, photoconversion provided a stable, nonfading DAB reaction product for light microscopy. In addition, at the electron microscopic level, it appeared that the photoconversion results in a homogeneously distributed, fine granular, dark, intracellularly located reaction product. With most of the retrograde tracers tested, photoconversion led only to staining of the cell bodies and the proximal portions of primary dendrites. Following photoconversion with intracellularly LY-filled neurons and cells labeled retrogradely with DiI, DiO, and 5,7-DHT, the reaction product was present throughout the cells, extending from the cell bodies into dendrites and dendritic appendices, and into axons. The high selectivity and methodological simplicity of photoconversion of DAB with fluorescent dyes into a stable, light and electron microscopical dense reaction product provide a promising alternative to classical neuroanatomical techniques and a new useful application of fluorescent neuronal tracers to light and electron microscopy. © 1993 Wiley-Liss, Inc.
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    Identification of synaptic interactions of intracellularly injected neurons in fixed brain slices by means of dual-label electron microscopy (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 31-42 
    Details
    ISSN: 1059-910X
    Keywords: Intracellular injection ; Fixed slice ; Dual-label immunocytochemistry ; Silver-gold enhancement ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The injection of the dye Lucifer Yellow (LY) into neurons in slices of fixed brain is used to associate cells displaying a particular dendritic geometry with a specific pattern of neuronal connectivity. In the present report we expand on this technique by combining it at the electron microscopic level with immunocytochemistry and/or degeneration for the study of synaptic relationships. As a model we use the projection neurons of nucleus accumbens. These neurons were retrogradely labeled in vivo with injections or a fluorescent tracer. Fast Blue, into the ventral mesencephalon. Using epifluorescent monitoring, these neurons were located in perfusion-fixed brain slices and intracellularly injected with LY. They were visualized in the light and electron microscope using a peroxidase-antiperoxidase immunocytochemical method. Certain afferent connections of these neurons were identified in the same tissue through the use of either dual-label immunocytochemistry or anterograde degeneration combined with a single-label immunoreaction. In the dual-label procedure, a silver-gold intensification of the diaminobenzidine (DAB) reaction product for the first antigen (LY) was contrasted with a nonintensified reaction product for the second antigen (tyrosine hydroxylase [TH]). Ultrastructurally, metallic gold particles appeared to be dispersed over the immunolabeled perikarya, dendrites, and, occasionally, axonal terminals of LY-injected neurons whereas the flocculent DAB reaction product was present in TH-containing axons and terminals. Following lesions of the ventral subiculum in the hippocampal formation, degenerating axon terminals were detected in nucleus accumbens along with immunoreacted, LY-injected neurons. The techniques outlined in this report should prove invaluable for the study of the synaptic interactions of identified neurons. They can be reliably reproduced with a high yield per experiment. © 1993 Wiley-Liss, Inc.
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    Application of environmental scanning electron microscopy in the development of detergents and personal products (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 424-428 
    Details
    ISSN: 1059-910X
    Keywords: Conventional scanning electron microscopy ; Adhesives ; Microcapsules ; Enamel ; Pellicle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Environmental scanning electron microscopy enables the observation of samples in their natural state with no preparation. Unilever Research employs this technology in the research and development of detergents and personal products with much success. The results presented are from various studies which demonstrate the versatility and utility of the technique. Vacuum sensitive samples such as microcapsules were successfully characterized, the efficacy of dental products was shown by monitoring an enamel surface before and after treatment, and the performance of an adhesive was assessed by dynamic experimentation. © 1993 Wiley-Liss, Inc.
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    Environmental scanning electron microscope (ESEM) evaluation of crystal and plaque formation associated with biocorrosion (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 429-433 
    Details
    ISSN: 1059-910X
    Keywords: Biocorrosion ; Sulfate-reducing bacteria ; Biofilm ; Desulfovibrio ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The biofilm attributed to Desulfovibrio vulgaris growing in the presence of ferrous metals was examined with an environmental scanning electron microscope. This novel microscope produced images of iron sulfide colloids and other iron containing structures that had not been reported previously. A plaque composed of iron sulfide enveloped the surface of the corroding metal while crystals containing magnesium, iron, sulfur, and phosphorus were present in the culture where corrosion was in progress. A structure resembling the tubercule found in aerobic corrosion was observed on stainless steel undergoing biocorrosion and the elements present in this structure included sulfur, iron, chloride, calcium, potassium, and chromium. © 1993 Wiley-Liss, Inc.
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    Morphology of nitric acid and water ice films (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 434-438 
    Details
    ISSN: 1059-910X
    Keywords: ESEM ; Surface reaction ; Polar stratospheric clouds ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ice films have been used to simulate stratospheric cloud surfaces in order to obtain laboratory data on solubilities and heterogeneous reaction rates. To obtain intrinsic uptake and surface reaction probabilities which can be applied to atmospheric models, it is necessary to carefully characterize these films. In the present study, environmental scanning electron microscopy (ESEM) is used to study thin films of both water ice and nitric acid ice near the composition of the trihydrate. The ices are formed by vapor deposition onto aluminum or borosilicate-glass substrates cooled to about 200 K. Micrographs are recorded during the deposition process and during subsequent annealing at higher temperatures. The results show that the ice films are composed of loosely consolidated granules, which range from about 1 to 20 μm in size at temperatures between 197 and 235 K. Cubic water ice is sometimes observed at 200 K and converts to the hexagonal form at slightly higher temperatures. The loose packing of the granules confirms the high porosities of these films obtained from separate bulk porosity measurements. Average surface areas calculated from the observed granule sizes range from about 0.2 to 1 m2 g-1 and agree with surface areas obtained by gas-adsorption (BET) analysis of annealed ice films. For unannealed films, the BET areas are about an order of magnitude higher than the ESEM results and imply that the unannealed ices contain microporosity which is lost during the annealing process. The present results have important implications for the extraction of intrinsic reaction probabilities from laboratory rate data. © 1993 Wiley-Liss, Inc.
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    Application of ESEM to environmental colloids (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 439-446 
    Details
    ISSN: 1059-910X
    Keywords: Microscopy ; Groundwater ; Pollution ; Radioactive waste ; Transport ; Remediation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Environmental colloids are toxic or radioactive particles suspended in ground or surface water. These hazardous particles can facilitate and accelerate the transport of toxicants and enhance the threat to humans by exposure to pathogenic substances. The chemical and physical properties of hazardous colloids have not been well characterized nor are there standard colloid remediation technologies to prevent their deleterious effects. Colloid characterization requires measurement of their size distribution, zeta potential, chemical composition, adsorption capacity, and morphology. The environmental scanning electron microscope (ESEM) by ElectroScan, Inc., analyzes particle sizes, composition, and morphology. It is also used in this study to identify the attachment of colloids onto packing or rock surfaces in our development of a colloid remediation process.The ESEM has confirmed the composition of groundwater colloids in our studies to be generally the same material as the surrounding rock. The morphology studies have generally shown that colloids are simply small pieces of the rock surface that has exfoliated into the surrounding water. However, in general, the source and chemical composition of groundwater colloids is site dependent. We have found that an ESEM works best as a valuable analysis tool within a suite of colloid characterization instruments. © 1993 Wiley-Liss, Inc.
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    Oil sorption behavior of various sorbents studied by sorption capacity measurement and environmental scanning electron microscopy (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 447-455 
    Details
    ISSN: 1059-910X
    Keywords: Cotton ; Milkweed ; Kapok ; Polypropylene ; Bicomponent fiber ; Biconstituent fiber ; Adsorption ; Absorption ; Capillary action ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Oil sorption capacities of various natural and man-made fibrous sorbents were compared in a simulated seawater bath containing oil. Natural sorbents such as milkweed, kapok, cotton, and wool showed higher sorption capacities than man-made sorbents such as polyester, polypropylene, viscose rayon, nylon 6, nylon 66, and acetate. Sorption capacities of the natural sorbents were over 30 g oil/g fiber. No definite advantages were observed using man-made bicomponent and biconstituent fibers over regular man-made fibers with respect to their sorption capacity.Analyses of sorption mechanisms using an environmental scanning electron microscope revealed that an oil deposit disappeared from the fiber surface after a certain time interval in milkweed, kapok, and cotton. This suggested that the sorption of oil in these fibers occurred through capillary action, probably due to their hollow lumens. Contrarily, adsorption, a surface phenomenon, would be the most prominent mechanism for oil sorption of wool fibers due to large amounts of surface wax, irregular scaly surfaces, and crimp. Effects of both adsorption and absorption were shown in the oil sorption of man-made fibers, depending upon the type and shape of the sorbent. Dumbbell-like oil deposits were seen on the fiber surface in certain oleophilic man-made fibers, because of a partial wetting of oil on the fiber surface. For some hydrophilic man-made fibers such as polyvinylalcohol and copolymer of isobutylene-maleic anhydride, the physical configuration of the fiber was a decisive factor in determining oil sorpton capacity of the sorbents. © 1993 Wiley-Liss, Inc.
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    Proposed healing and consolidation mechanisms of rock salt revealed by ESEM (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 456-464 
    Details
    ISSN: 1059-910X
    Keywords: Crushed rock salt ; ESEM ; Deformation ; Healing mechanism ; Consolidation mechanism ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The grain boundary healing behavior of crushed rock salt was mainly studied by employing the environmental scanning electron microscope (ESEM) to study the consolidation mechanism of rock salt backfill. Dedicated miniature round rock salt specimens were prepared for observation of the water trapping effect by using a cold stage in the ESEM to reach saturation conditions. Comparable high pressure pellets were prepared for measuring the crystal growth. Consolidation tests using materials made at different pressures and containing different moisture levels were conducted in order to construct the proposed mechanism. Direct observation of specimens in the ESEM resulted in viewing water trapped on the surface and the formation of a water meniscus between two particles. The concentration of brine at the grain boundary was observed as contributing to the amount of recrystallization. From aforementioned observations, a schematic drawing of the dissolution and recrystallization process may be redrawn. The amount of water therefore has a great effect on the consolidation of rock salt and is possibly due to the sliding, rotation, or crushing of the contact zone of the granular material. From such a study, tentative healing and consolidation mechanisms can be deduced. © 1993 Wiley-Liss, Inc.
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    Imaging fluid/solid interactions in hydrocarbon reservoir rocks (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 465-473 
    Details
    ISSN: 1059-910X
    Keywords: ESEM ; Liquid hydrocarbons ; Hydrocarbon reservoirs ; Clay minerals ; Chlorite ; Illite/smectite ; Calcite ; Fluid sensitivity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The environmental scanning electron microscope (ESEM) has been used to image liquid hydrocarbons in sandstones and oil shales. Additionally, the fluid sensitivity of selected clay minerals in hydrocarbon reservoirs was assessed via three case studies: HCl acid sensitivity of authigenic chlorite in sandstone reservoirs, freshwater sensitivity of authigenic illite/smectite in sandstone reservoirs, and bleach sensitivity of a volcanic reservoir containing abundant secondary chlorite/corrensite. The results showed the suitability of using ESEM for imaging liquid hydrocarbon films in hydrocarbon reservoirs and the importance of simulating in situ fluid-rock interactions for hydrocarbon production programmes. In each case, results of the ESEM studies greatly enhanced prediction of reservoir/borehole reactions and, in some cases, contradicted conventional wisdom regarding the outcome of potential engineering solutions. © 1993 Wiley-Liss, Inc.
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    Examples of evolution of microstructure in ceramics and composites (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 474-486 
    Details
    ISSN: 1059-910X
    Keywords: Coatings ; Copper thick films ; Crystallization ; Hydroxyapatite ; Propellants ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this article we describe a number of studies involving the direct observation of microstructural evolution. In general these investigations were carried out to establish the mechanistic paths involved. The materials studied range from fibers being evaluated for use in high-temperature ceramic composites to energetic materials used as propellants. In particular we discuss the room temperature imaging of materials difficult to image by conventional means and the use of the chamber atmosphere to influence microstructural evolution. Imaging of hydroxyapatite formed by chemical means is briefly described as an example of a difficult microstructure. Microstructural evolution during calcium aluminate cement hydration relies on the chamber atmosphere to control moisture loss from the hydrating specimens. In some instances microstructural evolution with heating occurred independently of the chamber atmosphere. Grain growth in PZT films formed by sol-gel processes depends strongly on temperature but does not appear to depend on the chamber atmosphere. This is also the case for the combustion of nitroamine propellants in that their combustion does not depend on access to an external source of oxygen. In other studies, the chamber atmosphere played an indirect role in determining microstructure. However, the mechanistic path driving microstructural evolution in copper-based inks used as conductive paths on electronic substrates is atmosphere dependent. These inks are formulated from copper powder, glass, and an organic binder, and the interaction of the binder with an oxidizing atmosphere allows it to be burned out before significant interaction occurs between the copper powder and the glass. Finally, the microstructural variations during the oxidation of structural composites at high temperature were used to allow assessments of their likely failure mechanisms. © 1993 Wiley-Liss, Inc.
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    Application of ESEM to fluxless soldering (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 493-502 
    Details
    ISSN: 1059-910X
    Keywords: Solder joining ; Ambient effects ; Solder oxidation ; Solder microstructure ; Solder morphology ; Atmosphere effects ; Solder ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ESEM is ideally suited to study soldering processes. We have used it to observe solder reflow and joining in ambient gases. It reproduces effects of atmospheric pressure reflow in a hot stage light microscope, but with much better clarity and depth of field. Compared to a regular SEM, the ESEM offers advantages of atmosphere control and ability to observe the solder samples without carbon or gold coating. These coatings could interfere with the oxidation/reduction reactions which occur at the solder/ambient gas interface. Very thin surface films, especially oxide layers dramatically influence the flow of liquid solder and the ability of solder to wet or join to another surface. Fluxless processes in particular are ideally suited for study in the ESEM. We have used the ESEM to observe dynamic fluxless soldering and have recorded events on videotape for later stop-action still pictures and slow motion photography. Examples of these processes are shown to illustrate the ESEM capability. Included are solder deformation structure, balling reflow of eutectic solder in hydrogen, balling reflow of eutectic solder in nitrogen, joining of two solder disks in nitrogen, and dynamic melting and freezing of an off-eutectic dendritic alloy. All of these are observed in the absence of flux. © 1993 Wiley-Liss, Inc.
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    Copper/Solder intermetallic growth studies (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 503-508 
    Details
    ISSN: 1059-910X
    Keywords: Intermetallic compounds ; ESEM ; SEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Copper samples, hot solder (eutectic) dipped and thermally aged, were cross-sectioned and placed in an environmental scanning electronic microscope (ESEM). While in the ESEM the samples were heated for ∼ 2.5 h at 170°C to stimulate the growth of additional Cu/Sn intermetallic compound. The intent of the study was to obtain a continuous real-time videotape record of the diffusion process and compare the observations to static SEM images reported to represent long-term, naturally aged intermetallic growth. The video obtained allows the observation of the diffusion process and relativistic growth phenomena at the Cu, Cu3Sn, Cu6Sn5, and solder interfaces as well as effects on the bulk Cu and solder. Effects contrary to earlier reports were observed; for example, growth rates of Cu3Sn were found to greatly exceed those of Cu6Sn5. © 1993 Wiley-Liss, Inc.
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    Patterns of intercellular connectivity in the mammalian cumulus-oocyte complex (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 125-133 
    Details
    ISSN: 1059-910X
    Keywords: Follicle cell ; Cumulus-oocyte-complex ; Transzonal processes ; Tubulin ; Actin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Electron and fluorescence microscopic techniques have been used in a complementary fashion to study the patterns of follicle cell-oocyte interactions within cumulus-oocyte-complexes of various mammals. The principal findings are: (1) two distinct types of transzonal processes exist that are distinguishable on the basis of cytoskeletal composition; (2) in some of the species examined (pig, goat, primate), corkscrew-shaped processes rich in tubulin, traverse the zona pellucida and are invaginated into the oocyte cortex; (3) actin-rich processes either ramify as a network at the outer surface of the zona pellucida or penetrate the zona and make contact with the oolemma in a species specific manner. These results are discussed with respect both to the need to employ complementary optical methods in assessing connectivity patterns within COC and to the possible role that extracellular matrix-cell interactions play in the homeostatic control of oocyte growth and maturation. © 1994 Wiley-Liss, Inc.
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    Concrete - pathology - secondary precipitations (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 179-180 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    Programmable freeze-substitution and cryo-embedding device (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 173-179 
    Details
    ISSN: 1059-910X
    Keywords: Cryoelectron microscopy ; Cryofixation ; Freeze-substitution ; Low temperature embedding ; Vitrification ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The construction and performance of a modular and fully controlable freeze-substitution device are described. The core of the device consists of a heavy brass block providing a large, stable thermal mass. The block is composed of two perforated plates and a base plate forming nine deep wells, which enable the concomitant substitution of several samples in various substitution fluids, and in large volumes. The wells are surrounded by an isometric network of tunnels through which either liquid nitrogen or hot air can flow. The isometric network enables heat transfer across short uniform distances throughout the entire block's volume, thus minimizing temperature gradients and differences. The temperature of the substitution fluid, rather than that of the metal block, is monitored by a programmable controller, enabling the presetting of any freeze-substitution regime. © 1993 Wiley-Liss, Inc.
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    Cytokines and growth factors in implantation (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 201-207 
    Details
    ISSN: 1059-910X
    Keywords: Cytokines ; Growth factors ; Blastocyst ; Implanatation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A variety of cell types at the blastocyst implantation site produce cytokines and growth factors that could play an important role in the implantation process. Furthermore, receptors for cytokines and growth factors have been detected on embryonic and trophoblastic cells. The purpose of the article is to review the published literature on the effect of cytokines and growth factors on implantation events, and to present recent data from our laboratory on effects of growth factors and cytokines on mouse blastocyst implantation events in vitro. © 1993 Wiley-Liss, Inc.
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    Immunohistochemical markers of uterine receptivity in the human endometrium (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 208-222 
    Details
    ISSN: 1059-910X
    Keywords: Immunohistochemistry ; Endometrium ; Implantation ; Biological markers ; Uterine receptivity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The factors responsible for the initial interaction between maternal and fetal epithelium leading to the establishment of pregnancy remain poorly understood. Temporal and spatial expression of specific endometrial peptides in response to ovarian steroids is thought to contribute to the development of a period of uterine receptivity, whereby the endometrium becomes hospitable to the implanting blastocyst. The failure to establish receptivity may account for a significant percentage of the cases of infertility in the female, especially affecting women with luteal phase deficiency, leiomyomata uteri, endometriosis, habitual abortion, and unexplained infertility. In addition, despite increasing global experience with advanced reproductive technologies, the majority of In Vitro Fertilization (IVF) attempts remain unsuccessful, most likely on the basis of implantation failure. In this article, we review the concepts involved in the study of uterine receptivity in the human, highlight potential immunohistochemical (IHC) markers that have recently been discovered, and discuss how IHC assessment of the endometrium is a potentially valuable method for the evaluation of the receptive endometrial state. Using this approach we have examined several new potential markers of uterine receptivity. Endometrial progesterone receptors and one of the integrin cell adhesion molecules appear to undergo changes in expression around the time of implantation, and may be sensitive indicators of the receptive state. Further, these markers are delayed in women with infertility and luteal phase deficiency. These studies illustrate the utility of IHC diagnosis for the evaluation of endometrial function. © 1993 Wiley-Liss, Inc.
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    Masthead (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070250401
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    Controlled-Depth and cross-section preparation techniques for transmission electron microscopy subsurface studies in metals (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 255-263 
    Details
    ISSN: 1059-910X
    Keywords: Thin film preparation ; TEM ; Austenitic stainless steel ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Thin film specimen preparation from bulk material at a controlled depth below the surface and cross-section thin film preparation for transmission electron microscope investigations of electrically conducting materials are described. Both techniques are illustrated by austenitic stainless steel, where they have been used complementary to each other for microstructural studies of subsurface ion irradiation damage. The advantages and limitations of both techniques are discussed. © 1993 Wiley-Liss, Inc.
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    Three-dimensional detection of the expression of intercellular adhesion molecule-1 (ICAM-1) in the high endothelial venule (HEV) of the rat lymph node (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 264-265 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    Endometrial adenocarcinoma (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 246-254 
    Details
    ISSN: 1059-910X
    Keywords: Adenocarcinoma ; Curettage ; Cytogenetics ; Endometrium ; FIGO staging ; Hormone receptors ; Hyperplasia ; Immunochemistry ; Oncogenes ; Prognostic factors ; Risk factors ; Screening tests ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Adenocarcinoma of the endometrium is the most common gynecologic malignancy in the United States, accounting for some 36,000 cases of invasive cancer each year. Hyperplastic lesions of the endometrium follow a continuum, with the risk of progression to carcinoma being related to the severity of the disorder. Risk factors associated with the development of adenocarcinoma include hyperplasia, obesity, menstrual abnormalities, diabetes, hypertension, prior pelvic irradiation, sequential oral contraceptive use, diet, and exogenous estrogen use. There is also some evidence of genetic predisposition, and some data indicating the possibility of specific genetic abnormalities and activation of oncogenes as factors determining the etiology of the disease. At this time there is no accepted screening test for endometrial carcinoma, though the role of immunochemistry techniques for screening and follow-up has just begun to be realized. Dilatation and curettage along with hysteroscopy remain the major means of diagnosis. A variety of prognostic variables including tumor cell type, histologic grade, depth of myometrial invasion, status of peritoneal cytology, presence of disease in preformed vascular spaces, presence of adnexal metastases, and presence of cervical involvement have been defined. Although the treatment plan for each patient must be individualized, the mainstay of treatment remains total abdominal hysterectomy with bilateral salpingo-oophorectomy. Metastatic and recurrent disease is usually treated with hormonal therapy and systemic chemotherapy. Radiation therapy like surgery in recurrent disease is only applicable for the treatment of local recurrences. © 1993 Wiley-Liss, Inc.
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    Characterization of agarose as an encapsulation medium for particulate specimens for transmission electron microscopy (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 267-275 
    Details
    ISSN: 1059-910X
    Keywords: Agarose encapsulation ; TEM specimen preparation ; Bacteria ; Yeast ; Mitochondria ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Agarose, agar, and gelatin were initially compared as encapsulation media for 3 structurally diverse particulate specimens: bacteria, yeast, and mitochondria. Agarose proved superior to both gelatin and agar for ease of handling and overall image quality (minimum background). All sample types exhibited high quality fixation and structural detail with no heat damage from the agarose medium. Based on this finding, we further characterized agarose encapsulation as affected by post-fixation, en bloc staining and resin type. Osmium tetroxide post-fixation, followed by en bloc uranyl acetate staining, could be performed without an increase in the electron density of the encapsulation medium. Agarose proved successful as an encapsulation medium regardless of the resin type or preparation protocol, thus providing flexibility in experimental design and excellent results over a range of variables. © 1993 Wiley-Liss, Inc.
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    Rapid primary microwave-glutaraldehyde fixation preserves the plasma membrane and intracellular structures of the protozoan Tritrichomonas foetus (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 286-290 
    Details
    ISSN: 1059-910X
    Keywords: Microwave fixation ; Freeze-fracture ; Electron microscopy ; Protozoan ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Tritrichomonas foetus, a pathogenic protozoan, was used as a model to analyse microwave-stimulated fixation as a procedure of preparation of biological samples for electron microscopy of thin sections and freeze-fracture replicas. Good preservation of the protozoan structure was achieved by microwave-stimulated fixation and Epon polymerization. The membrane structure, as visualized in freeze-fracture replicas, was well preserved. © 1993 Wiley-Liss, Inc.
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    Interactions between brain mitochondria and cytoskeleton: Evidence for specialized outer membrane domains involved in the association of cytoskeleton-associated proteins to mitochondria in situ and in vitro (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 233-261 
    Details
    ISSN: 1059-910X
    Keywords: Brain mitochondria ; Microtubules ; Neurofilaments ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The surface distribution of several proteins (porin, hexokinase, and two proteins associated with microtubules or actin filaments) on the outer membrane of brain mitochondria was analyzed by immunogold labelling of purified mitochondria in vitro. The results suggest the existence of specialized domains for the distribution of porin in the outer mitochondrial membrane. Similarities between the distribution of porin and the distribution of microtubule-associated proteins bound in vitro to mitochondria suggested that mitochondria and microtubules interact by binding microtubule-associated proteins to porin-containing domains of the outer membrane. This hypothesis was supported by biochemical studies on outer mitochondrial proteins involved in in vitro binding of cytoskeleton elements. In vitro interactions between mitochondria and microtubules or neurofilaments were analyzed by electron microscopy. These studies revealed cross-bridging between the outer membrane of mitochondria and the two cytoskeleton elements. Cross-bridging was influenced by ATP hydrolysis and by several proteins associated with the surface of mitochondria or with microtubules. In addition, unidentified proteins which were recognized by antibodies to all intermediate filaments subunits were associated either with the mitochondrial surface or with microtubules. This data suggest the participation of additional cytoplasmic proteins in the interactions between cytoskeleton elements and mitochondria. © 1994 Wiley-Liss, Inc.
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    Molecular and cellular mechanisms of mitochondrial nuclear division and mitochondriokinesis (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 220-232 
    Details
    ISSN: 1059-910X
    Keywords: Mitochondrial DNA ; Mitochondrial nuclear division ; Mitochondriokinesis ; Physarum polycephalum ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Our present understanding of mitochondrial division can be summarized as follows:Mitochondria contain a specific genome, synthesize their own DNA, and multiply semi-autonomously. Strands of mitochondrial DNA (mt-DNA) in the in vivo organelles of all eukaryotes are organized to form mitochondrial nuclei (nucleoids) (mt-nuclei) with specific proteins including a histone-like protein and transcription factors at the central region of the mitochondrion. We can easily observe the mt-nucleus in vivo mitochondria in various organisms such as fungi, algae, plants, and animals by using high-resolution epifluorescence microscopy. Therefore, the process of mitochondrial division can be clearly separated into two main events: division of the mt-nuclei and mitochondriokinesis analogous to cytokinesis.Mitochondria undergo binary division which is accompained by the division of the mt-nucleus. A remarkable characteristic of mitochondrial multiplication during the mitochondrial life cycle is that mitochondria can multiply the mt-chromosome by endoduplication until 50-100 copies are present. Mitochondria can then divide without mitochondrial DNA synthesis to eventually contain 1-5 copies of the mt-chromosome. This characteristic phenomenon can be observed during cell differentiation, such as during the formation of plasmodia and sclerotia of Physarum polycephalum and during embryogenesis and the formation of meristematic tissues in plants.The mitochondrial chromosome has a mitochondrial “kinetochore (centromere)” which is A-T rich and contains specific sequences such as topoisomerase binding sites, tandem repeats, and inverted repeats. A bridge of proteins may exist between the kinetochore DNA and membrane systems. Mitochondrial chromosomes can divide according to the growth of a membrane system between the kinetochores.Mitochondriokinesis progresses steadily along with mitochondrial nuclear division. As the membrane at the equatorial region of a mitochondrion contracts, the neck of the cleavage furrow narrows, and eventually the daughter mitochondria are separated. An actin-like protein may power mitochondriokinesis by separating the daughter mitochondria. In general, mitochondriokinesis occurs by contraction rather than by partition of the inner membrane. © 1994 Wiley-Liss, Inc.
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    Masthead (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070270401
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    Scanning tunneling microscopy I and scanning tunneling microscopy II, edited by R. Wiesendanger and H.-J. Güuntherodt. Springer Series in Surface Science #20 and #28, Springer Verlag, New York, 1992, 246 pp, $59.00 and 308 pp, $69.00 (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 268-268 
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    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    URL: http://dx.doi.org/10.1002/jemt.1070270307
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    Electron microscopy of the F1F0 ATP synthase: From structure to function (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 294-306 
    Details
    ISSN: 1059-910X
    Keywords: Cryo-electron microscopy ; Image analysis ; ATPase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The 1F0 ATP synthase is the large multisubunit complex which uses the proton gradient of energetically active membranes to synthesize ATP. While biochemical and genetic approaches have characterized the composition of the enzyme and elucidated many details of its mechanism and assembly, electron microscopy has been the tool of primary importance in determining the arrangement of the many subunits which comprise the F1F0. The highly cooperative catalytic mechanism is tightly coupled to transmembrane proton translocation in a separate and rather distant sector of the complex. An understanding of this intricate process and its control requires an appreciation of subunit interactions, starting with their locations relative to one another. Electron microscopy has provided most of the available structural information on the F1F0, and recent applications of cryo-electron microscopy have captured different functionally relevant configurations which may finally address longstanding questions about subunit rearrangement during the catalytic cycle. © 1994 Wiley-Liss, Inc.
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    Variations in mitochondrial ultrastructure and dynamics observed by high resolution scanning electron microscopy (HRSEM) (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 269-277 
    Details
    ISSN: 1059-910X
    Keywords: Cristae ; 3D structure ; Hepatocytes ; Fibroblasts ; Adrenal cortex ; Brown fat ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Rat adrenal cortex was processed for high resolution scanning electron microscopy (HRSEM) to confirm tubular cristae, reported by transmission electron microscopy to be present in cortex mitochondria. Mitochondria in several other tissue and cell types were also observed and their ultrastructure confirmed by using three-dimensional, stereo, high resolution scanning electron microscopy. The mitochondria in rat and human hepatocytes as well as human skin fibroblasts mitochondria proved to be long, up to 46 micrometers and branching, as compared to those in liver which were spherical in shape. Cold adapted brown fat cells were packed with mitochondria, these containing plate or shelf-like cristae. Branched, rat striated muscle mitochondria were observed to curve around contractile protein filament bundles. The muscle mitochondrial cristae were found to be both tubular and plate-like, within the same mitochondrion. The ratio of tubular cristae to plate-like cristae varied considerably between muscle mitochondria. In order to use ultrastructural changes in mitochondria for differential diagnosis, and because 3D reconstruction of mitochondria based on transmission electron microscopy serial sections is severely limited in resolution, it is imperative to first develop a correct understanding of tissue specific, normal mitochondrial ultrastructure based on three-dimensional, HRSEM methods. © 1994 Wiley-Liss, Inc.
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    The internal compartmentation of rat-liver mitochondria: Tomographic study using the high-voltage transmission electron microscope (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 278-283 
    Details
    ISSN: 1059-910X
    Keywords: Matrix ; Membrane ; SEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The three-dimensional organization of the internal compartments of conventionally fixed and embedded rat-liver mitochondria has been determined by tomographic reconstruction from tilt-series images collected on the Albany high-voltage electron microscope. The results indicate that the inner membranes of these organelles are predominantly tubular in the orthodox (expanded matrix) conformation, as previously suggested by scanning electron microscopy. In the condensed (contracted matrix) conformation, the intracristal space opens up into large irregularly shaped compartments which are connected to each other and to the external (intermembrane) space by tubes with approximately the same diameter (20 nm) as those observed in the orthodox state. These results raise several questions, in particular about the nature of the structural transitions that occur in the cristae during matrix expansion and contraction, and about the influence of inner-membrane shape on the diffusion of ions and metabolites between the intracristal and intermembrane compartments. © 1994 Wiley-Liss, Inc.
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    A morphological view on mitochondrial protein targeting (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 284-293 
    Details
    ISSN: 1059-910X
    Keywords: Mitochondrion ; Contact sites ; Protein translocation ; Ribosomes ; Import intermediates ; Receptor proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Mitochondrial protein targeting includes both intramitochondrial sorting of proteins encoded by the organellar genome and import and subsequent sorting of nuclear encoded precursor proteins. Only a few proteins are encoded by the mitochondrial genome and synthesized in the organellar matrix. These include predominantly inner membrane proteins that are perhaps co-translationally inserted into this membrane. Biochemical data suggest that insertion into the inner membrane may be confined to the inner boundary membrane. Ultrastructurally, however, a preferential association of ribosomes with either inner boundary or cristae membranes has not been established.The majority of the mitochondrial proteins are nuclear encoded and synthesized as precursors in the cytosol. Electron microscopic studies revealed that import of precursor proteins is generally confined to sites where both mitochondrial envelope membranes are closely apposed. In line with these observations, biochemical studies indicated that precursor proteins destined for the inner membrane or matrix have to interact with the energized inner membrane to allow complete passage of the precursor through the outer membrane. As a consequence, the mitochondrial envelope membranes have to be in close proximity at protein import sites.In isolated mitochondria distinct sites (designated as contact sites) exist where both envelope membranes are closely apposed and presumably stably associated. In situ, however, mitochondrial boundary membranes are in close proximity over large areas that cover almost the entire mitochondrial periphery. Consequently, the relative area of the mitochondrial surface, where both boundary membranes are in sufficient proximity for allowing protein translocation, is generally larger in situ compared to that in isolated organelles.Immunocytochemical localization studies showed a rather random distribution of components of the mitochondrial protein translocation machinery over the entire mitochondrial surface and not confined to contact sites.Based on these ultrastructral data and recent biochemical findings we propose that mitochondrial protein import sites are dynamic in nature and include relatively labile regions of close association of the boundary membranes. In vitro, however, mitochondrial protein import may preferentially take place at or near the presumably stable contact sites. © 1994 Wiley-Liss, Inc.
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    Proceedings of the fifth meeting of the great lakes electron microscopy affiliates, held in Indianapolis, IN, October 21-23, 1993 (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 350-354 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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    URL: http://dx.doi.org/10.1002/jemt.1070270409
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    Gastrulation in the mouse embryo: Ultrastructural and molecular aspects of germ layer morphogenesis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 301-328 
    Details
    ISSN: 1059-910X
    Keywords: Ectoderm ; Endoderm ; Mesoderm ; Egg cylinder ; Mouse embryo ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ultrastructural studies and lineage analyses of gastrulating mouse embryos have revealed that differnt morphogenetic tissue movements are involved in the formation of the three definitive germ layers. Definitive ectoderm is formed by epibolic expansion of the pre-existing progenitor population in the embryonic ectoderm. Formation of the mesoderm and the endoderm is initiated by cellular ingression at the primitive streak. The mesodermal layer is established by cell migration and cell sheet spreading, but the endoderm is formed by replacing the original primitive endodermal population. To this date, genes that are expressed during mouse gastrulation mostly encode cell surface adhesion or signalling molecules, growth factors and their receptors, and putative transcriptional factors. Their precise role during gastrulation remains to be investigated. © 1993 Wiley-Liss, Inc.
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    Masthead (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993) 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070260501
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    Immunohistochemistry of a human type I pneumocyte-associated protein in lung (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 357-365 
    Details
    ISSN: 1059-910X
    Keywords: Surfactant ; Squamous cells ; Plasma membrane ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A mouse monoclonal antibody to a human lung lavage protein was raised using proteins, with the potential ability to bind surfactant, as the immunogen. The proteins were isolated from cadaver lung lavage. The antibody was tested for its reactivity with lung and other organs. It reacted with type I pneumocytes and some of the nonciliated cells in the surface epithelium of distal bronchioles. Staining was also seen in the cells surrounding the glandular structures, superficial keratinocytes of the skin, endothelium, and nerve sheath cells. With the exception of bronchiolar cells, the stained cells have a squamous morphology, and this protein may serve as a marker or determinant of this characteristic of cells. In pathologic lungs some of the cells in air spaces with “bronchiolarization” of the epithelium exhibited staining for the protein. It could not be ascertained whether the stained cuboidal cells were reactive type II pneumocytes or distal bronchiolar cells. The intraalveolar material in pulmonary alveolar proteinosis did not show remarkable staining for the protein. Even though the protein is not unique to type I pneumocytes, it may serve as a marker for these cells in the study of their development and biology. © 1993 Wiley-Liss, Inc.
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    Interaction of lung surfactant protein a with alveolar macrophages (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 374-380 
    Details
    ISSN: 1059-910X
    Keywords: C1q ; Calcium ; Specific binding ; SP-A ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We analyzed the binding mechanism of human recombinant lung surfactant protein A (SP-A) to rat alveolar macrophages using anti-SP-A antiserum and protein A coated onto gold particles. Results were compared with our recent data on binding and uptake of SP-A-coated colloidal gold particles. The rationale for the current approach was to avoid any possible steric effects on SP-A binding to the cell surface. Binding of unlabeled SP-A depends on the presence of calcium ions in the medium and involves a mannose-specific mechanism. Binding is partly inhibited by the collagenase-resistent fragment of SP-A, representing mainly the globular part of SP-A. Taken together, these facts indicate binding of SP-A via the carbohydrate binding site on the globular region of SP-A. On the other hand, a partial inhibition of SP-A binding by fragments of C1q (representing the collagenous region of C1q) indicates a second binding site for SP-A by the collagen-like portion to the C1q receptor of macrophages. We conclude that two different mechanisms are probably involved in SP-A binding to alveolar macrophages. Specificity of the binding was shown with fluorescein-labeled SP-A. Binding was inhibited by an excess of unlabeled SP-A. Binding and uptake of SP-A are seen only with alveolar macrophages and not with other macrophage populations isolated from rat, such as liver macrophages (Kupffer cells), resident peritoneal macrophages, and peritoneal macrophages activated by Corynebacterium parvum. Therefore, binding sites for SP-A occur exclusively on alveolar macrophages. In addition, the intracellular Ca2+ concentration of the lung macrophages was determined by using the fluorescent dye fura-2/AM. Intracellular [Ca2+] increased immediately after addition of SP-A. This indicates immediate activation of macrophages by SP-A. © 1993 Wiley-Liss, Inc.
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    Ultrastructural features of alveolar epithelial cells in the late fetal pulmonary acinus: A comparison between normal and hypoplastic lungs using a rat model of pulmonary hypoplasia and congenital diaphragmatic hernia (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 389-399 
    Details
    ISSN: 1059-910X
    Keywords: Alveolar type II cells ; Surfactant ; Nitrofen ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The aim of this study was to describe and compare the ultrastructural features and functional maturity of alveolar epithelial cells in hypoplastic and normal fetal rat lungs. Pulmonary hypoplasia in association with congenital diaphragmatic hernia was induced in fetuses by administration of 2,4-dichlorophenyl-p-nitrophenylether (Nitrofen) to pregnant Sprague Dawley rats (100 mg on day 10 of gestation). Lung tissue of Nitrofen-exposed and control fetal rats aged 19-22 days (vaginal plug day 1, birth day 23) was embedded in Epon. Semithin (1 μm) toluidine blue-stained sections were examined by light microscopy; ultrathin sections (∼80 nm) were studied via transmission electron microscopy. In bronchoalveolar lavage fluid from control and Nitrofenexposed fetuses (day 22), phospholipid fractions and surfactant protein A content were measured semiquantitatively. On day 19 both control and Nitrofen-exposed lungs contained only cuboid alveolar epithelial cells; from day 20 there were cuboid, low cuboid, and thinner epithelial cells. The (low) cuboid cells contained large glycogen fields, some precursory stages of multilamellar bodies (MLBs), and just a few mature MLBs on day 19 and 20; smaller glycogen fields, more precursory stages, and more mature MLBs on day 21; and little or no glycogen but many precursory stages and mature MLBs on day 22. The thinner cells contained little or no glycogen and a few precursory stages of MLBs on days 20-22; very thin cells on day 22 contained neither glycogen nor any precursory stages of MLBs. MLBs and tubular myelin were seen in the lumens of future air spaces from day 20 onward. Nitrofen-exposed lungs differed from control lungs in that inclusion bodies (IBs) were less numerous in (low) cuboid alveolar cells on days 19 and 20, and more glycogen was seen on day 22. In addition intra- and extracellular “MLBs” in exposed lungs more often had an unusual appearance, i.e., a confluent structure and higher electron density. However, despite morphologic differences, there was no clear difference in phospholipid composition and SP-A content per mol phospholipid in bronchoalveolar lavage fluid. We conclude that morphologically hypoplastic lungs are less mature near term, without an apparent effect on surfactant composition. © 1993 Wiley-Liss, Inc.
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    Surfactant and inhaled particles in the conducting airways: Structural, stereological, and biophysical aspects (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 423-436 
    Details
    ISSN: 1059-910X
    Keywords: Surface forces ; Mucus ; Sol phase ; Airway macrophages ; Microscopy ; Fractionator ; Surface balance ; Video bronchoscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have investigated the displacement into the sol phase of inhaled particles deposited in the intrapulmonary conducting airways. Hamsters inhaled an aerosol of monodisperse polystyrene particles of 6 μm diameter. Their lungs were fixed by intravascular perfusion, and light and electron microscopy was used to study the epithelial coating. The surfactant film at the wall-air interface was investigated by measuring its surface tension. The number of particles retained was determined stereologically. In addition we investigated the displacement of spherical particles in vitro on a DPPC monolayer in a Langmuir-Wilhelmy surface balance and determined the surface tension in vivo in the horse trachea by video bronchoscopy, applying the droplet spreading method. We found that particles deposited onto a surfactant film were pulled into the aqueous subphase, and we concluded that surface forces due to the airway surfactant likely displace deposited particles into the periciliary fluid (sol phase). Comparing lungs fixed immediately after inhalation with lungs fixed 24 hr after inhalation revealed that 86% of the particles retained in the intrapulmonary conducting airways immediately after inhalation had been cleared within 24 hr. One-third of the particles of the lungs fixed immediately after inhalation was phagocytized. The combination of structural and stereological analyses with in vitro and in vivo measurements has led to new insights into the role of airway surfactant with respect to the fate of inhaled particles, which may have important consequences regarding the effects of hazardous particles, which may have important consequences regarding the effects of hazardous particles. These studies may also help to evaluate the deposition pattern and clearance of therapeutic particles, with important implications for their therapeutic use. © 1993 Wiley-Liss, Inc.
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    Role of alveolar macrophage chemotaxis and phagocytosis in pulmonary clearance responses to inhaled particles: Comparisons among rodent species (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 412-422 
    Details
    ISSN: 1059-910X
    Keywords: Complement activation ; Lung clearance ; Carbonyl iron particles ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Alveolar macrophages (AM) play an important role in clearing inhaled particles from the lung. The mechanisms through which macrophages identify particles that have been deposited in the alveolar regions is not well understood, although macrophage motility and phagocytic functions appear to be prerequisites for efficient clearance of inhaled materials. In previous studies, we assessed the mechanisms of macrophage-mediated clearance of inhaled particles using a rat model. In this regard, it appears that one mechanism by which rat alveolar macrophages are recruited to sites of particle or fiber deposition is through complement activation and consequent generation of chemotactic factors by the inhaled particulates. Whether this mechanism is operative in other rodent species remains an unanswered question. The current studies were undertaken to compare pulmonary clearance responses in several rodent species exposed to carbonyl iron (CI) particles. In vitro and in vivo pulmonary clearance responses were evaluated using one strain each of mouse, hamster, rat, and guinea pig. In vitro studies showed that hamster AM had the greatest phagocytic activity and that rat AM migrated best to complement-dependent chemotactic factors. Subsequently, groups of animals from each species were exposed to CI particles for 1 or 6 hr at aerosol concentrations of 100 mg/m3. Particle deposition patterns in the distal lung were nearly identical for all species, although enhanced numbers of CI particles were deposited on alveolar duct bifurcations of either rats or mice compared to hamsters, and particle deposition in guinea pigs was substantially lower. Time course studies showed that enhanced numbers of rat AM migrated to deposition sites and phagocytized particles, and this correlated with increased numbers and percentages of phagocytic macrophages recovered by lavage (P 〈0.01). In vivo phagocytic rates were the lowest in the mouse, and this correlated with reduced phagocytic rates in vitro. It is concluded form these studies that the rat may be the most efficient rodent species in clearing inhaled iron particles. In addition, it is conceivable that hamster AM are recruited to sites of particle deposition by a noncomplement-mediated mechanism. © 1993 Wiley-Liss, Inc.
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    A step section method for full histopathological assessment of carcinogen-affected target tissue during respiratory carcinogenesis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 466-471 
    Details
    ISSN: 1059-910X
    Keywords: Hamster lung cancer ; Experimental bronchial carcinogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Studies of carcinogenesis that are not limited to overt neoplasms but also involve evaluations of preneoplastic stages require histopathological assessment of the entire carcinogen-affected tissue so that the true nature and sequence of the progressive process can be determined. The customary serial sectioning approach achieves this goal, but at an inordinate logistic cost. In studies of hamster bronchial carcinogenesis, a step section method was compared to a quasi-random approach and to the customary serial section method. The step section method achieved the same diagnostic completeness as serial sectioning, but at a two orders of magnitude reduction in costs. © 1993 Wiley-Liss, Inc.
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    Acute lung injury during bacterial or fungal sepsis (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 444-456 
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    ISSN: 1059-910X
    Keywords: Escherichia coli ; Candida albicans ; Staphylococcus aureus ; Bacteremia ; Candidemia ; Cytokines ; TNF ; Adult respiratory distress syndrome ; Pulmonary edema ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We compared physiological and ultrastructural indices of acute lung injury (ALI) during septic shock caused by taxonomically diverse pathogens to distinguish ALI due to endogenous inflammatory mediators vs. microbial exotoxins or other factors. Conscious rats were infected i.v. with gram-negativeEscherichia coli(EC, serotype 055:B5), exotoxin-C producing gram-positiveStaphylococcus aureus (SA), or yeast-phaseCandida albicans(CA, a clinical isolate). Viable inocula of 1010 EC, 1010 SA, or 109 CA caused lethal shock in 〈24 h, but distinct types of ALI were noted after bacteria vs. fungi. Within 0.5 h of EC infection, leukocytes marginated in the lung vasculature; by death at 6-14 h, animals were hyperoxemic but not acidemic, and showed slight interstitial edema with increased wet/dry weight ratios (W/D = 5.22 ± 0.10, mean ± SE, vs. 4.86 ± 0.07 in controls, P 〈0.05). Similarly mild ALI occurred after 1010 SA. In contrast, within 0.5 h of CA infection, yeast were visible within lung intravascular leukocytes. By death at 6-12 h, CA animals showed hyperoxic acidemia and moderate ALI with capillary obstruction, interstitial hemorrhage, and elevated lung W/D (5.52 ± 0.13, P 〈0.05 vs. controls) associated with yeast-mycelial transformation. Prior neutropenia accelerated mortality and worsened ALI after CA, with hypoxemic acidemia, increased lung W/D (7.23 ± 0.34, P 〈0.05 vs. other groups), capillary occlusion, perivascular and alveolar hemorrhage, and septal disruption by mycelia. Bacteremia induced large increases in serum tumor necrosis factor-α (TNF) and interleukin-1α within 1.5 h, but these cytokines remained low in CA animals, even at death. Neither survival nor ALI after EC or CA was altered by pentoxifylline, which attentuated TNF production, or by cyclooxygenase inhibition with ibuprofen. Thus, overall ALI severity correlated with physiological indices of pulmonary function, but ultrastructural changes correlated better with pathogen type than circulating cytokine or eicosanoid mediators. Whereas lethal bacteremia induced early cytokinemia and mild ALI with or without bacterial exotoxins, moderate ALI apparently was mediated by fungal exotoxins during lethal candidemia, which worsened during neutropenia due to enhanced mycelial proliferation.© 1993 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070260512
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    Comparison of hexamethyldisilazane (HMDS), Peldri II, and critical-point drying methods for scanning electron microscopy of biological specimens (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 489-495 
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    ISSN: 1059-910X
    Keywords: Ultrastructure ; Tissue preparation ; Animal ; Plant ; Leaf ; Cuticle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Three different drying methods, critical-point drying (CPD), Peldri II, and hexamethyldisilazane (HMDS), were compared using representative animal( rat kidney, trachea, duodenum, lung, and red blood cells) and plant( leaves from ten species of monocotyledons and dicotyledons) specimens. All three drying methods produced identical results with animal specimens. Plant specimens showed signs of shrinkage regardless of which drying method was employed. The order of preservation quality from best to worst for leaves was CPD 〉 Peldri II 〉 HMDS, with the CPD method providing substantially better results in all but one case. Postfixation of leaves with osmium tetroxide resulted in poorer preservation in all instances. Peldri II caused complete extraction of leaf cuticular wax, while both both CPD and HMDS showed minimal extraction compared with samples air dried directly from acetone. These results indicate that HMDS provides a time-saving and inexpensive alternative to CPD for animal specimens. Plant specimens, particularly those containing cells with large central vacuoles, are adequately preserved only with the CPD method. In addition, postfixation with osmium should be avoided when processing plant specimens for scanning electron microscopy. © 1993 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070260603
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  • 98
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    Electron diffraction techniques, Vol. II, edited by J. M. Cowley. Oxford University Press and The International Union of Crystallography, Oxford, UK, 1993, 423 pp, $70,00 (1994)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 28 (1994), S. 79-79 
    Details
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070280113
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  • 99
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    Transcellular and paracellular pathways in lingual epithelia and their influence in taste transduction (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 196-208 
    Details
    ISSN: 1059-910X
    Keywords: Tight junctions ; Amiloride ; Na-K-ATPase ; Trigeminal ; Chorda tympani ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The lingual epithelium is innervated by special sensory (taste) and general sensory (trigeminal) nerves that transmit information about chemical stimuli introduced into the mouth to the higher brain centers. Understanding the cellular mechanisms involved in eliciting responses from these nerves requires a detailed understanding of the contributions of both the paracellular and transcellular pathways. In this paper we focus on the contribution of these 2 pathways to the responses of salts containing sodium and various organic anions in the presence and absence of amiloride. Electrophysiological recordings from trigeminal nerves, chorda tympani nerves, and isolated lingual epithelia were combined with morphological studies investigating the location (and permeability) of tight junctions, the localization of amiloride-inhibitable channels, and Na-K-ATPase in taste and epithelial cells. Based on these measurements, we conclude that diffusion across tight junctions can modulate chorda tympani and trigeminal responses to sodiumcontaining salts and rationalize the enhancement of taste responses to saccharides by NaCl. © 1993 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070260303
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  • 100
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    Morphometric and immunocytochemical assessment of fungiform taste buds after interruption of the chorda-lingual nerve (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 187-195 
    Details
    ISSN: 1059-910X
    Keywords: Atrophy ; Keratin ; Monoclonal antibodies ; Regeneration ; Reinnervation ; Tongue ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Unilateral interruption of the chorda-lingual nerve led to a loss of most epithelial axons and to the deterioration of fungiform taste buds in the anterior portion of the tongue of albino rats, mongolian gerbils, and golden hamsters. By three weeks after surgery the following percentages of fungiform taste buds had completely disappeared: 71% in gerbils, 28% in rats, and 26% in hamsters. Residual taste buds were classified into two groups: atrophic taste buds and taste bud remnants. Atrophic taste buds were smaller than normal and typically had no visible taste pore, although they retained the characteristic oval shape of a taste bud and numerous elongated cells. Taste bud remnants were non-oval fragments of taste buds with few elongated cells. Specific markers for elongated taste cells (monoclonal antibodies to keratin 19) confirmed that atrophic taste buds, as well as some taste bud remnants, had elongated taste cells. By 180 days after chorda-lingual nerve transection, 44% of rat fungiform taste buds had disappeared; morphometric analysis of the 311 residual taste buds established that 241 atrophic taste buds and 69 taste bud remnants were, respectively, 50% and 75% smaller than the average volume of 480 normal taste buds. The aggregate loss of gustatory tissue, calculated from the shrinkage of residual taste buds and the volume lost by the outright disappearance of many taste buds, was 88% for gerbils, 72% for rats, and 65% for hamsters. Evaluation in gerbils of the co-occurrence of taste buds and axons suggests residual taste buds were neurotrophically supported. Every gerbil fungiform papilla that lacked axons lacked a taste bud. Every fungiform papillae that had a residual taste bud had axons; axons were absent from 22% of empty fungiform papillae. Diminished numbers of gustatory neurotrophic axons could account for both the loss of fungiform taste buds and the reduced volume of residual taste buds. © 1993 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/jemt.1070260302
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