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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 274-280 
    ISSN: 1059-910X
    Keywords: Epoxy resin ; Acrylate and methacrylate resins ; Protein A-Gold ; Stereology ; Labeling intensity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The addition of 1% water to the epoxy resin Quetol increased the labeling intensity of the sample. The significant decrease of the curing temperature of the epoxy resin may assist in preservation of antigens. Water may also reduce the cross-linkage of the resin allowing more antigen to be available to the antibodies. The modified Quetol resin is an option for use in immunocytochemistry studies.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 288-297 
    ISSN: 1059-910X
    Keywords: SEM ; TEM ; Guinea pig ; Cochlea ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Inner ear tissue of the normal guinea pig was conductively stained (OTOTO-method) for SEM investigations. The Hensen's cells of the organ of Corti were removed using a micromanipulator inside the SEM. By this method atypical bodies of sensory and supporting cells were revealed in the apical turns of the cochlea. Atypical sensory cells showed great variations in size and shape. Several had no contact to Deiter's cells and no or only one nerve supply at their basal end. Atypical Deiter's cells showed alterations in shape and in the form of their phalangeal processes.Additionally altered parts of the organ of Corti were isolated by micromanipulation and embedded for correlative TEM-investigations.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 298-304 
    ISSN: 1059-910X
    Keywords: Fertilization ; Sperm-egg interaction ; Gamete fusion ; Egg activation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A method for correlative studies of early fertilization events that integrates techniques of intracellular electrophysiological recording, video-imaging, and electron microscopy is described. A key feature of the method is its ability to identify the fertilizing sperm and to record the moment of egg excitation. Since the site of gamete interaction is recognizable throughout all stages of preparation, difficulties associated with locating the site of fertilization and determining specimen orientation for microtomy and electron microscopic examination are eliminated. Virtually all samples yield useful information. An example of interacting gametes fixed 4 sec after initiation of the fertilization potential and serial sectioned is described. The method is applicable to systems other than fertilizing eggs when functional, temporal, and spatial relationships of individual cells need to be correlated with changes in ultrastructure.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 305-313 
    ISSN: 1059-910X
    Keywords: Cryofixation ; Development ; Fasciae adherentes ; Desmosomes ; Myocardium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Using the method of rapid freezing and freeze-substitution, the embryonic chick cardiac muscle was investigated by transmission electron microscopy. Initially, the intercellular junctional complexes (fasciae adherentes and desmosomes) were formed in close proximity to each other along a nearly straight line. Subsequently, the separation of fasciae from desmosomes took place to form intercalated discs. The cell membranes of fasciae adherentes were reinforced with highly interwoven fine fibrils at which myofibrils terminated. The intercellular space of fasciae was bridged with fine fibrillar structures seemingly connected by a thin line at their middle portions. In the intercellular space of desmosomes, central lamina and traversing filaments were clearly observed. The outer and inner leaflets of the desmosomal plasmalemma were asymmetrically differentiated; the outer leaflet was thinner than the inner leaflet. On the inner side of the cell membrane, an electron-lucent layer and a dense desmosomal plaque were observed. The latter structure had protrusions with less electron density towards the cytoplasmic side. Further inside, a meshwork of fine fibrils was seen along and toward which bundles of intermediate filaments ran. The results obtained with freeze-substitution appeared to provide more information than those with thin sections after conventional fixation or with replicas of chemically fixed/glycerinated or physically fixed/deep-etched materials.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 281-287 
    ISSN: 1059-910X
    Keywords: Creatine ; Creatine phosphokinase ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Creatine phosphokinase regenerates ATP from ADP using creatine phosphate. Isoenzymes of creatine phosphokinase are bound to certain cellular structures or are compartmentalized in areas of the cell, and this has been used as a basis for defining the role of these isoenzymes in energy metabolism. The M isoenzyme of creatine phosphokinase has been morphologically associated with the M-line of striated muscle in many species. In this present study the ultrastructural distribution and the relative concentration of the M form of creatine phosphokinase in human muscle tissue was determined using immunogold and electron microscopy. The M-line of the sarcomere, comprising only 3 - 4% of the sarcomere area, was found to contain over 20% of the total M isoenzyme signal of the entire sarcomere. This technique represents a quantitative, ultrastructural method to study the subcellular distribution of this isoenzyme. These data suggest that localized concentrations of M-CPK may be important for normal energy metabolism, and may also serve as a foundation for a better understanding of the relationship between abnormal creatine metabolism and the pathogenesis of neuromuscular disease.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 314-314 
    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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  • 7
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 315-316 
    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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  • 8
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992) 
    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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  • 9
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 318-332 
    ISSN: 1059-910X
    Keywords: RHEED ; Surface steps ; REM ; Surface defects ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Details of the technique of reflection electron microscopy (REM) are described. Step by step instruction is given on how to do it on an ordinary electron microscope. Also given are some specimen preparation techniques and strategy of REM investigation.
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  • 10
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 360-370 
    ISSN: 1059-910X
    Keywords: RHEED ; Bloch waves ; Green function ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Bloch wave equations for the multiple beam cases in reflection high energy electron diffraction (RHEED) are derived from the integral equation by forward- and back-scattering Green function operators. A linearization is achieved through separation in a forward- and a back-scattering component for each beam. This leads to a set of fundamental equations similar to the transmission case, but with a non-Hermitian matrix, and the beams may be entered as either forward-, back-scattered, or both. The number of beams needed to be included in RHEED calculations is thus reduced, and so are the computing time and computer space required. The systematic row case, corresponding to reflections from planes parallel to the crystal surface, is treated in detail and illustrated by calculations of dispersion surfaces and rocking curves for Au(001). Symmetry relations for the systematic row and between reciprocal rows are discussed and illustrated.
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  • 11
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 371-389 
    ISSN: 1059-910X
    Keywords: Bloch wave method ; Crystal surfaces ; Multislice ; Surface science ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: High energy electron reflection (HEER) is an important technique in surface science and uses the information carried by high energy electrons reflected from surfaces to study surface structures and surface electronic states. With the development of reflection high energy electron diffraction (RHEED), high energy electron microscopy (REM), and high energy electron energy loss spectroscopy (EEL) in surface science, the usefulness of HEER has been widely recognized and demonstrated. However, a stationary dynamical solution for an arbitrary surface for HEER has not been obtained yet.In this paper, some developments in understanding the dynamical theory of HEER, particularly in recent years, are reviewed: 1The introduction of the concept of current flow for a semi-infinite crystal model has removed the confusion around the wave points in the “band gap”.2The consistency between the Bloch wave and multislice in the Bragg case has verified the validity of the argument of current flow and led to the emergence of the BMCR method (Bloch wave + Multislice Combined for Reflection).3The failure of the Bloch Wave-Only solution (the BWO solution) on Au (110) surfaces in the Bragg case revealed by the BMCR method implies that previous BWO calculations in the Bragg case might be at fault.4The 2-D dependence of the electron wave fields and Picard iteration-like character of multislice calculation in the Bragg case has led to the emergence of an Edge Patching method in Multislice-mode-Only (the EPMO method). The new method yields an infinitely convergent stationary dynamical solution for an arbitrary surface.
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  • 12
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 390-405 
    ISSN: 1059-910X
    Keywords: Reflection electron microscopy ; Secondary electron imaging ; Field emission gun ; In situ REM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A review is given on the techniques and applications of high-energy reflection electron energy-loss spectroscopy (REELS) and reflection electron microscopy (REM) for surface studies in scanning transmission electron microscopes (STEM) and conventional transmission electron microscopes (TEM). A diffraction method is introduced to identify a surface orientation in the geometry of REM. The surface dielectric response theory is presented and applied for studying α-alumina surfaces. Domains of the α-alumina (012) surface initially terminated with oxygen can be reduced by an intense electron beam to produce Al metal; the resistance to beam damage of surface domains initially terminated with Al+3 ions is attributed to the screening effect of adsorbed oxygen. Surface energy-loss near-edge structure (ELNES), extended energy-loss fine structure (EXELFS), and microanalysis using REELS are illustrated based on the studies of TiO2 and MgO. Effects of surface resonances (or channeling) on the REELS signal-to-background ratio are described. The REELS detection of a monolayer of oxygen adsorption on diamond (111) surfaces is reported.It is shown that phase contrast REM image content can be significantly increased with the use of a field emission gun (FEG). Phase contrast effects close to the core of a screw dislocation are discussed and the associated Fresnel fringes around a surface step are observed. Finally, an in situ REM experiment is described for studying atomic desorption and diffusion processes on α-alumina surfaces at temperatures of 1,300 - 1,400°C.
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  • 13
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 406-412 
    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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  • 14
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 20 (1992), S. 413-425 
    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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  • 15
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 124-135 
    ISSN: 1059-910X
    Keywords: Pinealocyte ; Pineal-neuron ; Synapses ; Ribbon synapse ; Innervation ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Recent ultrastructural studies of neuronal-pinealocytic interconnections in the monkey pineal are reviewed. The pinealocytes in the adult monkey show almost all of the cytological specializations known in subprimate mammals. Adjacent pinealocytes are functionally coupled through ribbon synapses on cell bodies and gap junctions on cell bodies and cell processes. The pinealocytes receive direct synaptic contacts of nerve fibers with cholinergic terminal morphology. Nerve cells restricted to the central portion of the pineal receive synaptic contacts with more than three different morphologically defined types of nerve terminals. In addition to nerve terminals containing small clear vesicles or vesicles of pleomorphic morphology, a pinealocyte's terminal process containing the synaptic ribbon forms a true synaptic contact on the nerve cell body. The diversity of synapses on these nerve cells strongly suggests multiple origins of these neurons rather than a single peripheral parasympathetic origin. The possible involvement of pineal neurons in an intrinsic circuit that regulates the function of pinealocytes and integrates the neural input from the central as well as the peripheral nervous systems is discussed.
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  • 16
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    Microscopy Research and Technique 21 (1992), S. 85-115 
    ISSN: 1059-910X
    Keywords: Bats ; Chiroptera ; Pinealocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Pinealocytes are not only the principal cellular components of the pineal gland, but they are also the principal synthetic machinery of this enigmatical gland with highly diverse and often questionable empyreal roles assigned to it. Ultrastructural descriptions of pinealocytes belonging to some 70 species of mammals (a mere 2% or less of the over 4,200 mammalian species) have been summarized from the available literature with new observations on 12 species of chiropterans. Space limitation precluded any treatment of the supporting glia, neural elements, and the perivascular spaces. A detailed table lists nearly all mammalian species whose pineal ultrastructure has been investigated. Blanks in this table point to the necessity of studies on those particular groups. A tabular listing of unusual structures reported within the pinealocyte cytoplasm points out the impending experimental work on these species. Such studies using the latest techniques might provide clearer insights into the functional role of the pineal gland as an important and integral component of the neuroendocrine axis. Whereas sufficient structural information now exists on cytoplasmic organelles such as synaptic ribbons and spherules, annulate lamellae, subsurface cisterns, and the several types of synaptic arrangements seen in relation to the pinealocyte soma and its processes, the functional role of these structures in pineal synthetic processes remains to be elucidated.
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  • 17
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    Microscopy Research and Technique 21 (1992), S. 166-170 
    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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  • 18
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    Microscopy Research and Technique 22 (1992), S. 336-350 
    ISSN: 1059-910X
    Keywords: Bombyx mori ; Antheraea polyphemus ; Olfactory sensilla ; Cryofixation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The general morphology and methodological peculiarities of insect sensilla are briefly reviewed. The stimulus conducting pore-tubule systems of pheromone-sensitive sensilla of the silkmoths Bombyx mori and Antheraea polyphemus are described. Lipophilic tracers readily enter the hair lumen, while hydrophilic tracers do so only after prolonged extraction with lipid solvents and/or pronase. X-ray microanalysis demonstrates a high potassium content of the sensillum lymph; calcium was only found in the haemolymph above detection limit. Auxiliary cells rapidly take up radioactive leucine administered via the haemolymph. Antibodies against pheromone-binding protein of Antheraea polyphemus label the sensillum lymph of sensilla trichodea, but not of sensilla basiconica in A. polyphemus as well as in B. mori. The cytoplasm of auxiliary cells of the sensilla trichodea is also labelled. The results are discussed in context with present hypotheses on the role of sensillum lymph in stimulus transport and inactivation. © 1992 Wiley-Liss, Inc.
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  • 19
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    Microscopy Research and Technique 22 (1992), S. 351-371 
    ISSN: 1059-910X
    Keywords: Antheraea polyphemus ; Olfactory sensilla ; Differential mitoses ; Epidermal feet ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The olfactory organ of the silkmoth Antheraea polyhemus is the feathered antenna which carries about 70,000 olfactory sensilla in the male. It develops within 3 weeks from a leaf-shaped epidermal sac by means of segmental primary and secondary indentations which proceed from the periphery towards the centerline. During the first day post-apolysis, the antennal epidermis differentiates into segmentally arranged, alternating sensillogenic and non-sensillogenic regions. Within the first 2 days post-apolysis, the anlagen of olfactory sensilla arise from electron-dense mother cells in the sensillogenic epidermis. The axons of the developing sensilla begin to form the primary innervation pattern during the second day. The sensilla develop approximately within the first 10 days to their final shape, while the indentations are completed during the same period of time. The indentations are most probably driven by long basal extensions of epidermal cells, the epidermal feet. Primary indentations follow the course of segmentally arranged tracheal bundles and form the segments of the antenna. The secondary indentations follow the course of the primary segmental nerves which are reconstructed by this process. During the remaining time of development, the cuticle of the antenna and the sensory hairs is secreted by the epidermal and the hair-forming cells. © 1992 Wiley-Liss, Inc.
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  • 20
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    Microscopy Research and Technique 22 (1992), S. 403-406 
    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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  • 21
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    Microscopy Research and Technique 23 (1992), S. 200-200 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 22
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    Microscopy Research and Technique 24 (1993), S. 423-428 
    ISSN: 1059-910X
    Keywords: T system ; Muscle-tendon junction ; Lanthanum nitrate ; Stereomicroscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The 3-dimensional distribution of transverse (T) tubules at myotendinous junctions (MTJs) was studied by intermediate voltage (400 kV) electron microscopy of thick sections of rat vastus intermedius and chicken pectoralis muscles stained with lanthanum nitrate. Transversely oriented T tubules were seen to run at the level of A-I junctions (the rat vastus intermedius) and Z bands (the chicken pectoralis), but were absent from such levels adjacent to the end of MTJ processes. These tubules opened to the lateral wall of sarcolemmal infoldings of MTJs and to the lateral cell surface. Longitudinally running T tubules were seen to connect with the transverse T tubules and to open at the bottom of junctional folds. The lack of T tubules at the final sarcomeric regions seems to indicate that the terminal sarcomeric half in close proximity to MTJs may be activated from the sarcoplasmic reticulum which forms couplings with the MTJ sarcolemma and/or longitudinal tubules in the MTJ processes. © 1993 Wiley-Liss, Inc.
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  • 23
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    Microscopy Research and Technique 23 (1992), S. 157-172 
    ISSN: 1059-910X
    Keywords: N-CAM ; L1 ; AMOG ; J1 ; L2-HNK-1 ; Olfactory system ; Development ; Immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The localization of Ca++-independent cell adhesion molecules (CAMS) in the developing and mature olfactory epithelium and bulb is reviewed. The CAMs included in this article are the neural cell adhesion molecule (N-CAM), the 180 kD component of N-CAM (N-CAM 180), the embryonic form of N-CAM (E-N-CAM), L1 glycoproteins, J1 glycoproteins, and the adhesion molecule on glia (AMOG). In addition, the expression of the L2-HNK-1 carbohydrate epitope, shared by N-CAM, L1, J1 and myelin-associated glycoprotein (MAG) in the adult olfactory epithelium and bulb has also been documented. For the localization of these molecules at the light and electron microscopic levels, immunocytochemical techniques were used and are described in detail.During development and organogenesis, the olfactory system exhibits a pattern of CAM expression similar to the general pattern described for the developing nervous system. In the adult olfactory system, however, a significant retention of CAMs characteristic for developmental and morphogenetic processes, such as E-N-CAM, AMOG, as well as the high molecular weight components of J1 glycoproteins, can be observed. The retention of these embryonic features are most likely associated with the cell turnover and high plasticity of this system. Moreover, the predominance of N-CAM 180 with respect to other components of N-CAM, as well as the absence of the L2/HNK-1 carbohydrate epitope, are also particular traits of the primary olfactory system which could be associated with its exceptional properties. © 1992 Wiley-Liss, Inc.
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  • 24
    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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  • 25
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    Microscopy Research and Technique 24 (1993) 
    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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  • 26
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    Microscopy Research and Technique 24 (1993), S. 457-464 
    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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  • 27
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    Microscopy Research and Technique 23 (1992), S. 86-97 
    ISSN: 1059-910X
    Keywords: Accessory olfactory ; Nasal glands ; Odorant binding protein ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The vomeronasal and septal olfactory organs are two neurosensory structures in the mammalian nasal septum which are poorly understood relative to the main olfactory system. The vomeronasal organ is a paired, blind-ending tubular structure that opens rostrally into the nasal cavity in some species and into the incisive ducts in others. When present in mammals, the septal olfactory organ is an island of olfactory mucosa positioned such that it is in the primary air pathway in the caudal portion of the nasal cavity. Mammalian nasal glands, with a diverse histochemical and ultrastructural morphology, secrete a variety of substances onto the mucosal surface. One of these substances, odorant binding protein, localized in bovine nasal glands and lateral nasal glands of rodents may be important in the capture and conveyance of odorant molecules to olfactory receptors. The objectives of this paper are to present original data while reviewing the literature on the ultrastructure of vomeronasal and septal olfactory neuroepithelia, and of vomeronasal, bovine nasal, and lateral nasal glands. Nasal tissues from pigs, calves, and hamsters were prepared for electron microscopy. Neurosensory epithelia of the porcine vomeronasal organ and the hamster septal olfactory organ are similar to that described for the vomeronasal and septal olfactory organs of other mammals. Bovine nasal and rodent lateral nasal glands consist of subregions which differ morphologically; the most abundant acinar cell type in the bovine nasal gland contains lightly electron dense secretory granules while that of the rodent lateral nasal gland contains both small electron dense and large, electron lucent granules. The porcine vomeronasal gland contains numerous small, dense granules of a diverse morphology. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 29
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    Microscopy Research and Technique 23 (1992), S. 103-110 
    ISSN: 1059-910X
    Keywords: Olfaction ; Anosmia ; Pathology ; Nose ; Smell ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper presents electron-microscopic observations on biopsies of the olfactory mucosae of several classes of patients with smell disorders: 1) patients with loss of smell function following head injury (post-traumatic anosmics or hyposmics); 2) patients with loss of smell function following severe head colds and/or sinus infections (post-viral olfactory dysfunction, or PVOD); and 3) patients that have lacked smell function since birth (congenital anosmics). Of these, the traumatic anosmics' olfactory epithelia were quite disorganized; the orderly arrangement of supporting cells, ciliated olfactory receptor neurons, microvillar cells, and basal cells was disrupted. Although many somata of ciliated olfactory receptors were present, few of their dendrites reached the epithelial surface. The few olfactory vesicles present usually lacked olfactory cilia. The postviral anosmics, too, had a greatly reduced number of intact ciliated olfactory receptor neurons, and most of those present were aciliate. The post-viral hyposmics had a larger population of intact, ciliated olfactory receptor cells. In the seven cases of congenital anosmia studied, no biopsies of olfactory epithelium were obtained, indicating the olfactory epithelium is either absent - or greatly reduced in area - in these individuals. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 111-127 
    ISSN: 1059-910X
    Keywords: Bowman's glands ; Sustentacular cells ; Goblet cells ; Nerve terminalsa ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The mucus at the surface of the olfactory mucosa constitutes the milieu in which perireceptor events associated with olfactory transduction occur. In this review, the ultrastructure of olfactory mucus and of the secretory cells that synthesize and secrete olfactory mucus in the vertebrate olfactory mucosa is described. Bowman's glands are present in the olfactory mucosa of all vertebrates except fish. They consist of acini, which may contain mucous or serous cells or both, and ducts that traverse the olfactory epithelium to deliver secretions to the epithelial surface. Sustentacular cells are present in the olfactory epithelium of all vertebrates. In fish, amphibia, reptiles, and birds, they are secretory; in mammals, they generally are considered to be “nonsecretory,” although they may participate in the regulation of the mucous composition through micropinocytotic secretion and uptake. Goblet cells occur in the olfactory epithelium of fish and secrete a mucous product.Secretion from Bowman's glands and vasomotor activity in the olfactory mucosa are regulated by neural elements extrinsic to the primary olfactory neurons. Nerve fibers described in early anatomical studies and characterized by immunohistochemical studies contain a variety of neuroactive peptides and have several targets within the olfactory mucosa. Ultrastructural studies of nerve terminals in the olfactory mucosa have demonstrated the presence of adrenergic, cholinergic and peptidergic input to glands, blood vessels, and melanocytes in the lamina propria and of peptidergic terminals in the olfactory epithelium. The neural origins of the extrinsic nerve fibers and terminals are the trigeminal, terminal, and autonomic systems. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 180-184 
    ISSN: 1059-910X
    Keywords: Specimen preparation ; Cleaving ; Crystal structure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A small-angle cleavage technique has been developed that produces superior transmission electron microscope (TEM) samples of semiconductors and related materials. The technique involves back-thinning the sample to approximately 100 μm, then scribing a groove on this back face at a specified small angle to a standard cleavage plane. The sample is cleaved along this scribe line followed by cleaving along the standard cleavage plane to produce a thin wedged sample. Samples prepared by this method are characterized and compared with conventional and low-angle ion milled samples. The technique is illustrated, and the characteristic geometry of the cleaved sample is explained in terms of a simplified cleavage model. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 239-242 
    ISSN: 1059-910X
    Keywords: Braze alloys ; WC ; XTEM sample preparation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: An improved method for the preparation of cross-sectional thin foils of coated WC-Co samples for studies by analytical electron microscopy is described. A braze alloy is used to join the sections of the sample together and the resulting sample is stable during subsequent grinding, dimpling, and milling operations. Cross-sectional micrographs provide examples of the efficacy of this method. No microstructural alteration associated with the brazing operation was observed. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 230-238 
    ISSN: 1059-910X
    Keywords: High-resolution electron microscopy ; Twin boundaries ; Interphase boundaries ; Compositional order ; Reflex images ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The effects of crystal and beam tilt on high-resolution transmission electron microscope (HRTEM) images of planar coherent interfaces were investigated by multislice image simulations. It was found that a beam tilt of 0.5 Bragg angle (θB) was sufficient to introduce detrimental artifacts into most images of interfaces in crystals only 1/8ξ000 thick, while crystal tilt had a much smaller effect even for crystals 1ξ000 thick. Effects produced in HRTEM images of interfaces by crystal and beam tilt included the introduction of additional periodicities and loss of compositional detail across a boundary, translation of a boundary from its actual position, and apparent mismatch of atomic planes across a perfectly coherent interface. These results indicate that alignment of the electron beam parallel to the optic axis is critical for reliable HRTEM imaging of interfaces in materials. Techniques for obtaining accurate alignment are also discussed. © 1992 Wiley-Liss, Inc.
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    ISSN: 1059-910X
    Keywords: Peroxisomes ; Morphometry ; Protein A-gold ; Hypolipidemic drugs ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We describe the application of automatic image analysis for quantitative morphological studies of peroxisomes in rat liver. For automatic detection by light and electron microscopy peroxisomes must be stained with the alkaline DAB procedure for catalase. There is a good agreement between the results obtained by conventional morphometric techniques and by automatic image analysis of DAB-stained electron microscopic preparations. Moreover, the image analyzer may be used in conjunction with a light microscope for evaluation of semithin sections (1-0.25 μm), provided the section thickness factor is taken into consideration. This latter approach has proven highly efficient in estimation of peroxisome proliferation. The limitations of this method and the relevance of volume density as a reliable morphometric parameter for evaluation of peroxisome proliferation are discussed. In the second part of this study we present the application of image analysis for quantitation of alterations of individual peroxisomal enzyme proteins after treatment with bezafibrate in immunogold stained ultrathin sections. There is good agreement between the results of quantitative immunocytochemistry and Western (immuno) blot analysis of highly purified peroxisomal fractions. In our experience quantitative immunoelectron microscopy provides a versatile, highly sensitive, and efficient method for detection of modulations of various proteins in peroxisomes. Finally the limitations and prospects of quantitative immunocytochemistry for investigation of peroxisomal proteins are discussed. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 21 (1992), S. 292-299 
    ISSN: 1059-910X
    Keywords: Morphometry ; Histology ; Cytology ; Software ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper presents a snapshot view of the influence and direction of microcomputer technology for image analysis techniques in diagnostic pathology.Microcomputers have had considerable impact in bringing image analysis to wider application. Semi-automated tracing techniques are a simple means of providing objective data and assist in a wide range of diagnostic problems. From the common theme of reducing subjectivity in diagnostic assessment, an extensive body of research has accrued. Some studies have addressed the need for quality control for reliable, routine application.Video digitizer cards bring digital image analysis within the reach of laboratory budgets, providing powerful tools for investigation of a wide range of cellular and tissue features. The use of staining procedures compatible with quantitative evaluation has become equally important. As well as assisting scene segmentation, cytochemical and immunochemical staining techniques relate the data to biological processes.With the present state of the art, practical use of microcomputer based image analysis is impaired by limitations of information extraction and specimen throughput. Recent advances in colour video imaging provide an extra dimension in the analysis of multi-spectral stains. Improvements will also be felt with predictable increase in speed of microprocessors, and with single chip devices which deliver video rate processing. If the full potential of this hardware is realized, high-speed, routine analysis becomes feasible. In addition, a microcomputer imaging system can play host to companion functions, such as image archiving and transmission.With this outlook, the use of microcomputers for image analysis in diagnostic pathology is certain to increase. However, it is the software in both design and concept which ultimately governs the performance which can be achieved. Progress may be made by structured software techniques, by application of mathematical principles, or by use of expert systems for data or image interpretation. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 21 (1992), S. 300-314 
    ISSN: 1059-910X
    Keywords: Digital perimeter ; Digital area ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Quantitative estimations of perimeter and area from digitized video images, and the application of these features in morphometry, are discussed. Estimations from manual tracings via interactive peripherals and from chain codes are addressed. Topics discussed are calibration, determination of vertical and horizontal pixel resolution, effects of tracing jitter, and for chain codes, the spatial quantization scheme representation of the digital contour. Finally, new perimeter estimators for 4-connected and 8-connected chain codes for non-unity pixel aspect ratio are presented with simulation results.
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    Microscopy Research and Technique 23 (1992), S. 264-274 
    ISSN: 1059-910X
    Keywords: CNS ; Light microscopy ; Electron microscopy ; Cerebral cortex ; Nerve cell ; Impregnation method ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes the early stages of impregnation by the Golgi rapid method in sections and blocks of brain tissue. Aldehyde-fixed and potassium dichromate-treated sections of cerebral cortex were placed on glass slides and coverslipped. The dichromate solution was then replaced by a silver nitrate solution, and events taking place in the section were monitored and time-lapse recorded until the impregnation was interrupted and the sections subsequently prepared for electron microscopy. The tissue blocks, fixed and chromated in the same way, were placed into a silver nitrate solution for 30 min to 24 h and the progress of impregnation compared with the results obtained in the sections on the glass slides.Two basic modes of impregnation were observed, apparently in direct relation to the process of crystallization of silver chromate: crystals of silver chromate growing directly from the surface of the tissue into the nerve cell via its transected plasma membranes, and microcrystalline precipitate of silver chromate spreading into the nerve cell from nucleation centres dispersed in the tissue. The precipitate grows inside the cell as in a preformed channel until the cell has been filled. If the nucleation begins extracellularly, the precipitate extends into the narrow intercellular gaps. Electron microscopy showed that the crystalline precipitate consisted of multilamellar formations containing dense coalesced granules that did not cross plasma or endocellular membrane boundaries. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 275-288 
    ISSN: 1059-910X
    Keywords: Golgi-electron microscopy ; Gold substitution ; Chemical reduction ; Stabilization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Direct electron microscopy of nervous tissue stained with the Golgi impregnation method is unsatisfactory because the cytoplasm of the cell bodies and processes of the impregnated neurons are completely filled with a compact precipitate of electron dense silver chromate. This precipitate entirely obscures the cytological details of the impregnated neurons. Because of its solidity and instability in aqueous solutions, the silver chromate is also a source of inconvenience during the preparation of the ultrathin sections.This review summarizes methods that have been developed with the aim of replacing the Golgi precipitate in CNS neurons with a more convenient electron dense material - for example, heavy metal salts or metallic particles. Conversion of the precipitate into a stable electron dense marker is done before the material is embedded for electron microscopy. The methods include lead, gold, and bromide substitution, treatment with ammonia, direct chemical reduction into metallic silver, and photoreduction of the silver chromate into silver through irradiation with ultraviolet light. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 289-305 
    ISSN: 1059-910X
    Keywords: Golgi-EM procedure ; Identified neurons ; Synapses ; Neuronal taxonomy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The Golgi-electron microscope technique has opened new avenues to explore the synaptic organization of the brain. In this article, we shall discuss basic methodological principles necessary to analyze axonal arborizations with this combined technique. To illustrate the applications of the method, we shall review the forms and distribution of the synapses in which the axonal arborizations of local cortical interneurons engage. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 324-333 
    ISSN: 1059-910X
    Keywords: Golgi impregnation ; Golgi-electron microscopy ; Neuronal connectivity ; Neural network ; Synaptic connection ; Dendritic spine ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The combined light and electron microscopic analysis of Golgi-impregnated neural tissue is a potent tool for determining the connectivity of neural networks within the brain. In the experimental paradigms commonly applied in these studies, the Golgi-impregnated neurons are typically examined as the postsynaptic neuronal components. The structural characteristics and the pattern of distribution of their synaptic connections with other groups of identified neurons are analyzed. Due to the high power of resolution of the Golgi-electron microscopic technique, the ultrastructural analysis of Golgi-impregnated neurons can be expanded to elucidate activity-dependent structural alterations in their cytoarchitecture. These structural alterations can then be correlated under different physiological conditions with changes in the functional efficacy of the subcellular neuronal components. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 23 (1992), S. 306-323 
    ISSN: 1059-910X
    Keywords: Section Golgi impregnation ; Cholinergic synapses ; Neuronal specificity ; Neural transplantation ; Slice culture ; Neuronal plasticity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this study the Golgi/electron microscopy (EM) technique has been used for an analysis of the fine structure, specific synaptic connections, and differentiation of neurons in the hippocampus and fascia dentata of rodents. In a first series of experiments the specific synaptic contacts formed between cholinergic terminals and identified hippocampal neurons were studied. By means of a variant of the section Golgi impregnation procedure, Vibratome sections immunostained for choline acetyltransferase, the acetylcholine-synthesizing enzyme, were Golgi-impregnated in order to identify the target neurons of cholinergic terminals in the hippocampus. It could be shown with this combined approach that cholinergic septohippocampal fibers form a variety of synapses with different target structures of the Golgi-impregnated and gold-toned hippocampal neurons. In this report cholinergic synapses on the heads of small spines, the necks of large complex spines, dendritic shafts, and cell bodies of identified dentate granule cells are described. The variety of cholinergic synapses suggests that cholinergic transmission in the fascia dentata is a complex event.Next, the Golgi/EM technique was applied to Vibratome sections that contained retrogradely labeled neurons in the hilar region of the fascia dentata following horseradish peroxidase (HRP) injection into the contralateral hippocampus. With this combined approach some of the hilar cells projecting to the contralateral side were identified as mossy cells by the presence of retrogradely transported HRP in thin sections through these Golgi-impregnated and gold-toned neurons. Our findings suggest that the mossy cells are part of the commissural/associational system terminating in the inner molecular layer of the fascia dentata. They are mainly driven by hilar collaterals of granule cell axons that form giant synapses on their dendrites.Finally, the Golgi/EM procedure was used to study the differentiation and developmental plasticity of hippocampal and dentate neurons in transplants and slice cultures of hippocampus. Under both experimental conditions, the differentiating neurons are deprived of their normal laminated afferent innervation but develop their major cell-specific characteristics including a large number of postsynaptic structures (spines). As revealed in thin sections of gold-toned identified cells, all these spines formed synapses with presynaptic boutons suggesting sprouting of the transplanted and cultured neurons, respectively.Altogether, the present report demonstrates the usefulness of the Golgi/EM technique, particularly of the section impregnation procedure, for a variety of studies requiring the identification of individual neurons at the ultrastructural level. © 1992 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 40-45 
    ISSN: 1059-910X
    Keywords: Reverse hemolytic plaque ; In situ hybridization ; mRNA ; Pituitary lactotroph ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have developed an assay that allows one to monitor gene expression in and peptide secretion from individual cells. By combining the reverse hemolytic plaque with in situ hybridization, investigators can quantitate simultaneously the level of gene expression and the level of secretion of a peptide. The method can be used in any system in which an appropriate antibody for the reverse hemolytic plaque assay and probes complementary to the mRNA of interest are available. It can be used to monitor the level of mRNA and secretion of the peptide product, or expression of one gene and the secretion of another peptide. In this paper we will describe the major steps of the method. We have used the pituitary lactotroph as a model to demonstrate the power of this technique. However, we believe that this method may be an important approach to answer many questions regarding the cellular and molecular mechanisms that regulate the coupling of peptide secretion and gene expression at the single cell level. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 61-67 
    ISSN: 1059-910X
    Keywords: Morphometry ; Quantitative autoradiography ; mRNA ; Supraoptic nucleus ; Oxytocin ; Nested factorial design ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: One serviceable feature of in situ hybridization is its potential for assessing relative levels of mRNA in specific regions of tissues and organs. To determine its efficacy as a quantitative technique, we applied a nested factorial design to a multifactorial experiment. Estimates of the magnitude of variance components then allowed an assessment of variation over samples of sections from the same tissue source, variation in label over 2 anatomical sites within the same section of tissue, as well as experiment-to-experiment variation. We found ∼ 51% of the total variance arose from experiment-to-experiment variation, while ∼ 21% of the total variance was due to variation in autoradiography grain density over neurons in the same brain region. Rat-to-rat variation accounted for approximately 11%. About 10% of the variance was due to variation between sections of tissue that were derived from the same tissue source and were hybridized in the same hybridization experiment. Variation between 2 homologous, bilaterally located brain regions located on the same tissue section (the right and left supraoptic nucleus), accounted for ∼ 5% of the total variance. The remaining unaccounted error variance was ∼ 2% of the total variance. Since an expected change in cellular content of a particular mRNA was observed as a function of experimental treatment, results suggest in situ hybridization is a useful quantitative method. Findings also indicate, however, the importance of experimental design: the use of multiple samples of tissue from the same tissue source, a sufficient number of tissue sources (animals, batches of cultured cells) to account for variations in sample sources, and the need to assess experiment-to-experiment and rat-to-rat variations. Results suggest the utility of analyzing the data of in situ hybridization experiments from the perspective of experimental design. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 46-60 
    ISSN: 1059-910X
    Keywords: Vasopressin ; Vasoactive intestinal polypeptide ; Gastrin releasing peptide ; GABA ; In vitro ; In situ hybridization histochemistry ; Electron microscopy ; Circadian pacemaker ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Suprachiasmatic nuclei (SCN) from hypothalami of postnatal rats were maintained for 18-39 days in vitro as organotypic slice explants. Neuronal subtypes containing vasopressin (VP), vasoactive intestinal polypeptide (VIP), gastrin releasing hormone (GRP), and GABA were immunocytochemically identifiable in these cultures. In situ hybridization histochemistry was compatible with these SCN slice explant cultures, and mRNA encoding for VP was detected bilaterally within these nuclei. After 18 days in vitro, both VP mRNA and VP immunoreactivity increased from levels present on postnatal days 4 (the earliest age from which the explanted tissue was derived) to levels typical of adult SCNs. In contrast, the GRP expression remained low, characteristic of early postnatal animals and far lower than adult levels. This suggests that the developmental cues or programs necessary for enhanced VP expression are maintained in these cultures, while those affecting GRP expression are absent or inhibited. VIP-containing neurons were numerous in the cultures. Culture slices appeared healthy, and similar numbers and distributions of identifiable neurons within the SCN were observed, whether or not the slices were grown in the presence of serum. EM analysis revealed that the SCN in vitro is composed of tightly packed neurons, processes, and abundant synapses containing both clear and dense core vesicles, closely resembling the SCN in vivo. Vasopressinergic neuronal somata contained extensive Golgi systems and labeled secretory granules, the latter organelle being present also within processes and synaptic terminals. GABA-immunopositive processes and synaptic profiles were abundant, with labeling occurring particularly over secretory vesicles and mitochondria. This slice culture system effectively maintained much of the intrinsic organization and cellular components of the SCN for long periods in vitro and should be an excellent model system for studying the intrinsic molecular mechanisms and extrinsic cues which regulate neuronal phenotype in this circadian pacemaker. Published 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 78-84 
    ISSN: 1059-910X
    Keywords: Lentivirus ; HIV ; AIDS ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In situ hybridization (ISH) for HIV is an arduous, demanding means of detecting viral genetic material in cells and tissues. Good ISH requires broad technical skills and devotion to controls for every step of the process as well as a critical eye when interpreting results. ISH may be used to detect HIV in three ways: by hybridizing to viral RNA, by hybridizing to proviral mRNA being produced for virion packaging, and by hybridizing to proviral DNA in the cytoplasm or integrated in the nucleus of an infected cell. Here we discuss the technical considerations involved and the problems encountered in using ISH to study the pathobiology of HIV infection. Published 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 68-77 
    ISSN: 1059-910X
    Keywords: Female cell nuclei ; Xa ; CISS ; Confocal microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: It is a widely held belief that the inactive X-chromosome (Xi) in female cell nuclei is strongly condensed as compared to the largely decondensed active X-chromosome (Xa). We have reconsidered this problem and painted X-chromosome domains in nuclei of subconfluent, female and male human amniotic fluid cell cultures (46, XX and 46, XY) by chromosomal in situ suppression (CISS) hybridization with biotinylated human X-chromosome specific library DNA. FITC-conjugated avidin was used for probe detection and nuclei were counterstained with propidium iodide (PI). The shape of these nuclei resembling flat ellipsoids or elliptical cylinders makes them suitable for both two-dimensional (2D) and three-dimensional (3D) analyses. 2D analyses of Xi- and Xa-domains were performed in 34 female cell nuclei by outlining of the painted domains using a camera lucida. Identification of the sex chromatin body in DAPI-stained nuclei prior to CISS-hybridization was confirmed by its colocalization with one of the two painted X-domains. In 31 of the 34 nuclei the area AXi for the inactive X-domain was smaller than the area AXa for the active domain (mean ratio AXa/AXi = 1.9 ± 0.8 SD, range 1.0-4.3). The signed rank test showed a highly significant (P 〈 .0001) difference both between AXa and AXi and between the ratios r(Xa) and r(Xi), calculated by dividing the maximum length L of each X-domain by its maximum width W. In most nuclei (26/34) we found r(Xa)〉r(Xi) demonstrating a generally more elongated structure of Xa. For 3D analysis a confocal scanning laser fluorescence microscope (CSLFM) was used. Ten to 20 light optical sections (PI-image, FITC-image) were registered with equal spacings (approx. 0.4 μm). A thresholding procedure was applied to determine the PI-labeled nuclear and FITC-labeled X-domain areas in each section. Estimated slice volumes were used to compute total nuclear and X-domain volumes. In a series of 35 female nuclei most domains extended from the top to the bottom nuclear sections. The larger of the two X-chromosome domains comprised (3.7 ± 1.7 S.D.)% of the nuclear volume. A mean ratio of 1.2 ± 0.2 SD (range 1.1-2.3) was found for the volumes of the larger and the smaller X-domains in these female nuclei. In a series of 27 male amniotic fluid cell nuclei the relative X-chromosome domain volume comprised (4.0 ± 2.6 S.D.)%. These findings indicate that differences in the 3D expansion of active and inactive X-chromosome domains are less pronounced than previously thought. A current model suggests that chromosome domains consist of a compact core surrounded by loosely coiled outer chromatin fiber loops. The latter fraction may be considerably larger in Xa- as compared to Xi-domains. We suggest that the interactive outlining procedure used in the 2D analyses included the loosely structured domain periphery more accurately, while the threshold algorithm applied to light optical sections delineated the more compact core of the domains, leading to smaller and more similar volume estimates of Xa and Xi. Present limitations of nuclear and chromosome domain volume measurements using confocal laser scanning microscopy are discussed. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 48
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    Microscopy Research and Technique 25 (1993), S. 91-105 
    ISSN: 1059-910X
    Keywords: Endometrial maturation ; Ovarian steroids ; Hormonal interrelationships ; Life and Medical Sciences ; Cell & Developmental Biology
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  • 49
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    Microscopy Research and Technique 25 (1993), S. 346-349 
    ISSN: 1059-910X
    Keywords: Water-soluble substrate ; Replacement of film ; Ion beam thinning ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A preparational method was developed solving the problem of cross-sectional TEM preparation of thin films and layer systems deposited onto water-soluble substrates. The technique is based on the replacement of the sample onto steady substrate, followed by mechanical and ion beam thinning. Cross-sectional TEM micrographs of Ag and Ag/Ag2Se layers are shown presenting the efficiency of this novel technique. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 43-66 
    ISSN: 1059-910X
    Keywords: Retina ; Intracellular staining ; Horseradish peroxidase ; Electron microscopy ; Cone ; Horizontal cell ; Bipolar cell ; Amacrine cell ; Ganglion cell ; Neurotransmitter ; Synaptic plasticity ; Spinules ; Rhodamine ; Double labelling ; Postembedding immunohistochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A variety of intracellular recording and staining techniques has been used to establish structure-function and, in some cases, structure-function-neurochemical correlations in fish, turtle, and cat retinae. Cone photoreceptor-horizontal cell connectivity has been studied extensively in the cyprinid fish retina by intracellular staining with horseradish peroxidase (HRP) and subsequent electron microscopy. The available data suggest that horizontal cell dendrites around the ridge of the synaptic ribbon are postsynaptic, whilst finger-like extensions (“spinules”) of lateral dendrites function as inhibitory feedback terminals. An interesting feature of this inter-action is its plasticity: the feedback pathway is suppressed in the dark and becomes potentiated by light adaptation of the retina.Intracellular recordings and stainings of ganglion cells in both turtle and cat retinae have been possible. Prelabelling of ganglion cells by retrograde transport of rhodamine from the tectum allows ganglion cells to be stained under visual control, and their synaptic inputs determined by electron microscopy. Such studies have been extended to double labelling by using autoradiography or postembedding immunohistochemistry to identify the neurotransmitter content of the labelled cell and/or the neurotransmitter(s) converging upon it. It is envisaged that further applications of intracellular staining followed by double- or even triple-labelling will continue to enhance greatly our understanding of the functional architecture of the vertebrate retina. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 181-182 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 25 (1993), S. 185-186 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 25 (1993), S. 188-188 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 25 (1993), S. 189-200 
    ISSN: 1059-910X
    Keywords: Trophoblast ; Immunodeficient mice ; Extracellular matrix ; Cytokines ; Lymphokine-activated killer cells ; Abortion ; Murine pregnancy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The metrial gland develops in the uterus of many rodent species during normal pregnancy. It is a maternally-derived tissue that contains stromal and vascular elements plus a population of large cells, striking in their light microscopic appearance due to the presence of numerous cytoplasmic granules. These cells, which have become known in mice and rats as granulated metrial gland (GMG) cells, are derived from bone marrow precursors and recent work suggests they are a subset of lymphocytes belonging to the natural killer (NK) cell lineage. The functions of GMG cells during normal gestation have not been clearly defined. In vitro, GMG cells have been shown to produce cytokines and their cytokine profile is altered upon addition of medium containing the T cell growth factor interleukin-2 (IL-2). GMG cell granules contain the cytolytic protein perforin but GMG cells have a very limited capacity to kill in vitro unless they have been stimulated by IL-2 or interferon-gamma. Histological study of GMG cells has suggested they preferentially associate with fetal trophoblast. Since trophoblast appears resistant to immune lysis, except by IL-2-activated effector lymphocytes, and because resorbing murine embryos become infiltrated by lytic cells of the NK cell lineage, it is important to establish whether GMG cells are activated by pregnancy-associated events to play a major lytic role in vivo. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 85-102 
    ISSN: 1059-910X
    Keywords: Cerebellum ; Mesodiencephalon ; Spiny appendages ; Glomerulus ; Gap junction ; Rebound activation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper reports on the detailed morphology of inferior olivary neurons in the cat following electrophysiological examination, intracellular injection with horseradish peroxidase, and gamma aminobutyric acid (GABA) immunocytochemistry.The activity of olivary cells was recorded intracellularly in vivo and their response to mesodiencephalic stimulation was tested. In a number of cases their response to stimulation of the contralateral superior cerebellar peduncle was also tested. Mesodiencephalic stimulation resulted in monosynaptic, and superior peduncle stimulation in disynaptic activation of cells in the medial accessory and principal olivary subdivisions. Rebound olivary activity was usually only found after mesodiencephalic stimulation.Light microscopic investigation of osmicated and Araldite embedded Vibratome sections was facilitated considerably when performing the osmication in a glucose solution. Peroxidase labeled olivary cells, like that earlier described for Golgi-impregnated material, possess a complex globular dendritic geometry. Especially, and unlike Golgi material, the abundance of exceptionally long and complex spiny appendages could be appreciated. The axons usually stemmed from first order dendrites and did not give rise to recurrent axon collaterals.The ultrastructural analysis of this material, mainly from serial sections, was combined with postembedding GABA immunohistochemistry. In this way, GABAergic as well as non-GABAergic profiles were studied in conjunction with HRP labeled cellular elements.The GABAergic terminals usually contained pleomorphic vesicles and made symmetrical synapses whereas non-GABAergic terminals nearly always formed asymmetrical synapses and contained round or oval vesicles. Most, if not all, HRP labeled spiny appendages were incorporated in glomeruli. A particular spiny appendage may contribute more than one spine head to a glomerular core, which, on average, consisted of spiny elements of six different neurons. A glomerular core is surrounded by approximately the same amounts of GABAergic and non-GABAergic boutons. Also, all spiny appendages, and most of their individual spine heads, are contacted by GABAergic as well as non-GABAergic boutons. Spiny appendages on the axon hillock may be incorporated in dendritic glomeruli, however, most synapses with the hillock were made by GABAergic boutons.The combined physiological and morphological observations imply that (1) the cerebellar nucleican excert an excitatory influence on inferior olivary neurons through a mesodiencephalic relay, (2) the GABAergic nucleo-olivary input seems to be capable of diminishing the oscillatory tendencies of olivary neurons, and (3) the mesodiencephalic (non-GABAergic) and cerebellar (GABAergic) input may subserve a timing function since these inputs systematically impinge upon the same olivary spines. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 106-112 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 24 (1993), S. 113-130 
    ISSN: 1059-910X
    Keywords: Astrocytes ; Schwann cells ; Ensheathing cells ; Nerve entry zone ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This article provides a detailed description of the glial cell types in the nerve fiber layer of the main olfactory bulb during embryonic development, in adult mammals, and at the nerve entry zone of the first cranial nerve. In adult mammals, the glial cell types of the olfactory nerve fiber layer include intrafascicular ensheathing cells, which have the exclusive role of ensheathing the olfactory axons in both the PNS and CNS, and interfascicular astrocytes, which occupy the spaces between adjacent olfactory fascicles. The ensheathing cells are particularly interesting because they possess a mixture of Schwann cell and astrocytic phenotypic features, are more likely to be of placodal than of CNS origin, and have the exclusive role of forming the glia limitans at the PNS-CNS transitional zone. It is proposed that one important function of ensheathing cells is to modulate the growth of olfactory axons within the CNS; this modulation is probably mediated by selective cell adhesion molecules, extracellular matrix molecules, and chemotropic agents. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 131-141 
    ISSN: 1059-910X
    Keywords: Cell-cell interaction ; Neural recognition ; Glomerulus ; Olfactory receptor axons ; Axon guidance ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: How are the axonal projections of olfactory and vomeronasal receptor neurons to the olfactory bulb formed during development? How are the primary olfactory axonal connections functionally organized? With progress in molecular biological techniques and histochemical methods, it became possible to study cellular strategies and molecular mechanisms which guide the primary olfactory axons of the main and accessory olfactory systems to the target glomeruli in the bulb. In addition, new methodologies have begun to elucidate various subsets of the primary olfactory axons with distinctive central connections. The aim of the present paper is to review (1) the characteristic organization of the projection of the primary olfactory axons, (2) projection patterns of histochemically defined subsets of primary olfactory axons, and (3) information on molecules expressed by the surface membrane of the primary olfactory axons. This knowledge gives insight into the functional organization of the primary olfactory axon projection, which is indispensable for understanding signal processing in the olfactory system. This knowledge also underscores the notion that the primary olfactory axon projection provides an excellent model system in which to study axonal guidance and the formation of specific synaptic connections. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 157-167 
    ISSN: 1059-910X
    Keywords: Information processing ; Odors ; 2-Deoxyglucose ; Voltage-sensitive dyes ; Mouse ; Salamander ; Odor response modules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A number of different recording methods have shown that odorants elicit patterns of neuronal activity widely distributed across cells of the olfactory receptor epithelium, olfactory bulb, and piriform cortex in the vertebrate olfactory system. These findings suggest that the physicochemical properties of odorant molecules are processed by distributed coding mechanisms activated in parallel in olfactory circuits in order to characterize a single, “monomolecular” odorant. These findings also suggest that the response patterns seen at higher levels are set up by differential responses in peripheral receptor cells of the olfactory epithelium. One requirement for understanding the details of this proposed encoding scheme is correlation of odor-generated patterns with the components of these circuits. In this paper, results from 2-deoxyglucose and voltage-sensitive dye studies suggest that certain components of these responses may relate to patterns established in reproducibly identifiable aggregates of bulbar cells. These findings are consistent with previous observations suggesting that columnar groups of periglomerular, mitral/tufted and granule cells, oriented perpendicular to the laminae of the bulb, are functionally related to one another. Such cell groups or modules, when activated in parallel, could serve as building block components of the complete ensemble response. According to this hypothesis, different sets of such modules would be activated with different odorant stimuli and modules could be shared to the degree to which the physicochemical properties of the different stimuli overlap. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 168-172 
    ISSN: 1059-910X
    Keywords: Charge dissipation ; Absorption ; Attenuation ; Electron beam ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: It has been determined that, in the normal range of aluminium coating thicknesses used to remove charge from non-conducting specimens in the electron microscope, no detectable influence on the elemental signals obtained in X-ray microanalysis is observed. This is in contrast to a previous report (Hopkins et al., J. Electron Microsc. Tech., 18:176-182, 1991) of a reduction in elemental signal with increasing aluminium coating thickness. An explanation of errors in the previous interpretation is provided. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 24 (1993), S. 142-156 
    ISSN: 1059-910X
    Keywords: Sensory processing ; Olfactory coding ; Olfaction ; Odor stimulation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Complete understanding of the role of the mammalian main olfactory bulb in sensory processing has remained elusive despite many detailed studies on its anatomy and physiology. Several lines of recent evidence viewed in the context of earlier knowledge have provided new insights into the bulbar mechanisms of olfactory coding. The output cells of the olfactory bulb receive a localized olfactory nerve input and interneuronal input via dendrodendritic synapses on distinct sets of dendrites. The spatial arrangement of granule cell contacts on output cell basal dendrites suggests that lateral inhibitory interactions may occur between neighboring output cells. The input from olfactory receptor cell axons to the bulb also has spatial order, but does not represent a precise map of the receptor surface. Recent studies with antibodies and lectins suggest that different groups of axons from chemically similar receptor cells collect into certain glomeruli, even if the axons originate from cells that are not contiguous in the mucosa. Electrophysiological studies have begun to explore the participation of spatially organized circuits in olfactory processing. The degree to which neighboring output cells respond similarly to odor stimulation, for example, depends on the distance between the cells, with those further apart showing complementary responses. Also, a single output cell can show 2 or more different temporal response patterns when different odors are presented. Intracellular recordings indicate that these responses are shaped by IPSPs. Electrical stimulation during such recordings shows that some mitral cells are excited by nerve inputs close to their glomerular tufts, while they are inhibited by nerve inputs to other parts of the bulb. Finally, recordings from granule and periglomerular cells indicate their potential in mediating components of output cell odor responses. These considerations suggest that the olfactory bulb performs a spatially based analysis on the information coming from the receptor cells. While the spatial organization of the olfactory bulb is probably not faithfully represented in the projections to the olfactory cortex, bulbocortical projections are not random. The fact that spatial factors exist at each of these levels in the olfactory system must be considered in developing models of central olfactory processing. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 26 (1993), S. 301-328 
    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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    Microscopy Research and Technique 26 (1993) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 357-365 
    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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    Microscopy Research and Technique 26 (1993), S. 374-380 
    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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  • 66
    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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    Microscopy Research and Technique 25 (1993), S. 465-473 
    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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    Microscopy Research and Technique 25 (1993), S. 474-486 
    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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    Microscopy Research and Technique 27 (1994) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 27 (1994), S. 46-60 
    ISSN: 1059-910X
    Keywords: Salivary gland ; Tumor ; Differentiation ; Classification ; Experimental studies ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Electron microscopy has a limited role in the diagnosis of primary salivary gland tumors, although it can be helpful in metastatic lesions of possible salivary gland origin. The diversity of subtypes in salivary gland tumors, as well as the range of histomorphology within any one subtype, is unparalleled in any other human tumor. This and their relative infrequency causes diagnostic problems for pathologists. Ultrastructural techniques have been of major importance in determining the inter-relationship of these tumors for classification purposes, revealing the subtle variations in common cellular differentiation pathways, determining the organization of tumor cells, and displaying the importance of extracellular matrix materials in establishing diagnostic criteria for each of the many subtypes. Electron microscopy has also been valuable in non-neoplastic salivary gland disease and has an increasing role in experimental studies involving tissue from human and animal salivary parenchyma. © 1994 Wiley-Liss, Inc.
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    Microscopy Research and Technique 25 (1993), S. 503-508 
    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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    Microscopy Research and Technique 27 (1994), S. 125-133 
    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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    Microscopy Research and Technique 26 (1993), S. 92-92 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: No Abstruct.
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    Microscopy Research and Technique 26 (1993) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 93-93 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 489-495 
    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.
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    Microscopy Research and Technique 26 (1993), S. 496-512 
    ISSN: 1059-910X
    Keywords: TEM ; Sectioning techniques ; Specimen preparation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Epoxy resins are the principal embedding media for the preservation of tissues to be sectioned and examined by transmission electron microscopy. Their primary advantages are good ultrastructural preservation, little or no shrinkage, ease of sectioning, and reasonable stability in the electron beam. However, epoxy resins also have disadvantages; namely, some are toxic, they may mask antigenic sites to a greater extent than do some other embedding resins, and they do not penetrate tissues as well as less viscous embedding formulations. Some unusual characteristics may also be revealed, for example, as shrinkage of organelles, as problems in poststaining sections, and as movement of tissue elements within the block and section. Some of the properties of epoxy resins are discussed in this report. © 1993 Wiley-Liss, Inc.
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  • 78
    ISSN: 1059-910X
    Keywords: Mercox ; Vascular wall ; Resin casting ; Polymer casting ; Vascular corrosion casting ; Vascular replica/SEM method ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We used intravital microscopy of small intestine and pancreas in order to show dynamic interactions between vascular wall and undiluted Mercox, because previous studies of ours have shown that Mercox diluted with monomeric methylmethacrylate penetrates cells in the vascular wall. Scanning and transmission electron microscopy were used to show three-dimensional pathways and correlating tissue structures, which cannot be identified in vivo. The microvascular diameters were not altered when the vasculature was flushed with saline/dextran solution using perfusion pressures between 70 and 140 mm Hg, but, in circumscribed areas, contraction of vascular wall was observed immediately after Mercox injection. This phenomenon was carried out by endothelial cells; pericytes were never present at the site of constrictions. Extravasation, i.e., leakage of the resin into the surrounding tissue, occurred in circumscribed areas regardless of the applied perfusion pressure. The resin also filled routes, which were not perfused with blood before casting. Scanning microscopy of corresponding specimens showed flattened cast channels, with impressions of valves and endothelial cell nuclear imprints characteristic of lymphatics. These results show that undiluted Mercox is a stimulus for vascular cellular components and that it changes the vascular wall permeability, resulting in extravasation and filling of lymphatics. Transmission electron microscopy showed that large vessels were homogeneously filled with resin and that cellular structures were not infiltrated with Mercox. Cut sections of the gold-coated surface of casts showed grooves up to 20 nm wide, suggestive of minimal deformation, while the abluminal surface of the metal film was almost smooth. Another proof of minimal deformation of undiluted Mercox casts is that the diameter of vessels was not altered during and after polymerization. Obtained casts are not fragile, as are casts of diluted Mercox, and phase separation does not occur, which would result in penetration of the cells in the vascular wall. For these reasons, the use of undiluted Mercox is recommended. Mixing 10 ml Mercox with 1 g catalyst resulted in complete polymerization within 5.5-7 min. This mixture can be used for casting biological specimens. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 26 (1993), S. 524-524 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 27 (1994) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 27 (1994), S. 198-219 
    ISSN: 1059-910X
    Keywords: Mitochondria ; Organelles ; Microtubules ; Fluorochromes ; Fission ; Fusion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Mitochondria are semi-autonomous organelles which are endowed with the ability to change their shape (e.g., by elongation, shortening, branching, buckling, swelling) and their location inside a living cell. In addition they may fuse or divide. These dynamics are discussed. Dislocation of mitochondria may result from their interaction with elements of the cytoskeleton, with microtubules in particular, and from processes intrinsic to the mitochondria themselves. Morphological criteria and differences in the fate of some mitochondria argue for the presence of more than one mitochondrial population in some animal cells. Whether these reflect genetic differences remains obscure. Emphasis is laid on the methods for visualizing mitochondria in cells and following their behaviour. Fluorescence methods provide unique possibilities because of their high resolving power and because some of the mitochondria-specific fluorochromes can be used to reveal the membrane potential. Fusion and fission often occur in short time intervals within the same group of mitochondria. At sites of fusion of two mitochondria material of the inner membrane, the matrix compartment seems to accumulate. The original arrangement of the fusion partners is maintained for some minutes. Fission is a dynamic event which, like fusion, in most cases observed in vertebrate cell cultures is not a straightforward process but rather requires several “trials” until the division finally occurs. Regarding fusion and fission hitherto unpublished phase contrast micrographs, and electron micrographs have been included. © 1994 Wiley-Liss, Inc.
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    Microscopy Research and Technique 27 (1994), S. 262-267 
    ISSN: 1059-910X
    Keywords: Thick films ; Low-angle ion milling ; Differential sputtering rate ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A new preparation method permits the production of large-area, electron-transparent, transmission electron microscopy (TEM) specimens in cross section of free-standing, thick, multilayered structures. Such production often has been difficult in the past because of large chemical differences between the component layers in the multilayer. This difference usually results in a large difference in thinning rates between the layers. A unique combination of electroplating, lapping, dimpling, and low-angle ion milling is a successful and reproducible technique for producing high-quality TEM specimens of these complex materials. Procedures and results presented here are for a 304 stainless-steel/copper multilayer having a repeat period of 20 nm and a total thickness of 20 μm. © 1994 Wiley-Liss, Inc.This article is a US Government work and, as such, is in the public domain in the United States of America.
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  • 84
    ISSN: 1059-910X
    Keywords: Brain mitochondria ; Microtubules ; Neurofilaments ; Life and Medical Sciences ; Cell & Developmental Biology
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    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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    Microscopy Research and Technique 27 (1994), S. 220-232 
    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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    Microscopy Research and Technique 26 (1993), S. 171-176 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 182-183 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 180-181 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 184-185 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 26 (1993), S. 177-179 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Natural Sciences in General
    Notes: This device consists of a siphon system and 5 to 10 grid disks, modified from the previous mode (Chen, 1973), for large numbers of grids with ultrathin sections. This method improves the ease of assembling grids onto the grid disk and also requires much less stain solution. This system only takes 5 min for one single stain washing, at a maximum of 100 grids, and also avoids stain contamination. The grid disk can also be used for immunocytochemistry work and for critical point drying of grids with biological specimens.
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    Microscopy Research and Technique 26 (1993), S. 196-208 
    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.
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    Microscopy Research and Technique 26 (1993), S. 187-195 
    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.
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    Microscopy Research and Technique 26 (1993), S. 209-224 
    ISSN: 1059-910X
    Keywords: Cell types ; Ultrastructure ; Mammalian ; Ninth nerve ; Dense-cored vesicles ; Papilla ; Rabbit ; Trophic ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The fine structure of the taste buds of circumvallate papillae of two strains of mice was studied by electron microscopy. Mice anesthetized with ketamine were perfused through the heart with a double aldehyde mixture in cacodylate buffer and the tissues embedded in Epon. Semi-serial sections were employed. The morphology and relationships of cell types are consistent with the majority of descriptions of mammalian taste buds served by the ninth cranial nerve. Cells of type II are particularly well documented, as the stages in their origin, maturation and degeneration could be followed. Significant differences, however, relate to cell type I. These cells contain large dense-cored granules, contrasted with the more irregular and somewhat larger dark granules of the type I cells in the rabbit. These granules do not produce a dense homogenous product for the pore, as seen in the rabbit. Rather the pore substance consists of small, empty vesicles in a diffuse dark matrix. These granules are only moderately larger than the dense-cored vesicles of the type III cells. All features of the type III cell were demonstrated, although no instance of a complete cell was seen in any section. No significant differences were noted between the two strains of mice. Intimate proximity of a nerve to a cell nucleolus, suggestive of a trophic pathway, is illustrated. © 1993 Wiley-Liss, Inc.
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    Microscopy Research and Technique 26 (1993), S. 225-230 
    ISSN: 1059-910X
    Keywords: Gustation ; Neurotransmitters ; Ultrastructure ; Amphibia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The presence of glutamate immunoreactivity (glu-IR) in the nerve fibers of the mudpuppy taste bud was investigated by electron microscopy. Pre-embedding staining with avidinbiotin-peroxidase complex (ABC) and post-embedding staining with 5 mm colloid gold conjugates were used separately to identify immuno-stained structures. We have found the following: 1) the majority of the nerve fibers innervating the mudpuppy taste bud are unmyelinated; 2) about 85% of nerve fibers located at the base of the taste bud and about 60% of the nerve fibers located between the taste cells show glu-IR by pre-embedding staining; 3) there is a preferential staining of the glu-IR in the nerve fibers of the mudpuppy taste bud; and 4) the distribution of the colloidal gold particles in the nerve fibers is 1.5 to 2 times denser than that of the staining in the connective tissue background or cellular profiles of taste cells. From the distribution and pattern of the nerve fibers obtained in the thick and thin sections, we conclude that the mudpuppy taste bud is innervated by glutamate-containing unmyelinated nerve fibers. © 1993 Wiley-Liss, Inc.
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  • 96
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    Microscopy Research and Technique 26 (1993), S. 231-244 
    ISSN: 1059-910X
    Keywords: Chorda tympani ; Glossopharyngeal nerve ; Spinal trigeminal nucleus ; Taste ; Salivatory nuclei ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The distribution of acetylcholinesterase (AChE), NADH dehydrogenase (NADHd), and cytochrome oxidase (CO) was determined in the nucleus of the solitary tract (NST) in the golden hamster. Histochemical staining was compared to cytoarchitectonic subdivisions of the NST (Whitehead: J. Comp Neurol. 276:547-572, 1988) and to terminal fields of primary afferents of the nerves that innervate the tongue. These three histochemical methods resulted in differential staining patterns within the NST that were related to certain subdivisions. Transganglionic transport of horseradish peroxidase (HRP) was used to determine the central projections of the chorda tympani (CT), the lingual branch of the trigeminal (L-V), and the lingual-tonsilar branch of the glossopharyngeal nerves (L-IX). Alternate or the same brain sections were processed to reveal transported HRP, and NADHd or AChE levels. Increased staining of the neuropil with NADHd and AChE was coincident with the dense part of the afferent terminal fields of all three nerves in the NST and the laterally adjacent dorsomedial part of the spinal trigeminal nucleus. CO showed this pattern only for the most rostral part of the CT field. The densest AChE staining coincided with gustatory afferent terminal fields. The histochemical staining facilitated the interpretation of the organization of the NST. For example, at caudal levels of the gustatory NST, it is suggested that taste processing is localized predominately in the medial part of the rostral central, and somatosensory processing in the rostral lateral subdivision. AChE or NADHd staining should facilitate studies of connections, topography, and neuroplastic changes of the gustatory NST. © 1993 Wiley-Liss, Inc.
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  • 97
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 28 (1994), S. 125-140 
    ISSN: 1059-910X
    Keywords: Glycosaminoglycans ; Heparan sulfate ; Dermatan sulfate ; Cuprolinic blue ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The application of cationic probes for the ultrastructural detection of proteoglycans in basement membranes is reviewed. Proteoglycans are highly negatively charged macromolecules due to their glycosaminoglycan side chains. The interaction of cationic probes with proteoglycans is of an electrostatic nature. Methods are discussed to increase the specificity of probes for proteoglycans. The use of phthalocyanin-like dyes such as Cuprolinic blue, according to the critical electrolyte concentration method, results in a selective staining of proteoglycans. Enzymatic or chemical digestions, however, should be done to validate the proteoglycan nature of the dye-positive granules/filaments, and to establish the class of proteoglycan. The value of cationic probes in basement membrane research on development and pathology is discussed. The potential for deducting molecular information from the ultrastructural appearance of stained proteoglycans is indicated. © 1994 Wiley-Liss, Inc.
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  • 98
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 28 (1994) 
    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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  • 99
    ISSN: 1059-910X
    Keywords: Time-resolved fluorescence ; Fluorescence microscopy ; Europium chelates ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In the present study europium chelates were introduced as alternative fluorescent labels for microscopy and their effect on enhanced autofluorescence caused by the glutaraldehyde fixative was investigated. Glutaraldehyde fixation was used to stabilize the cells for a permanent mount after the immunocytochemical reaction. The europium signal in time-resolved fluorescence microscopy was shown to be free of autofluorescence when strong cross-linking fixation with glutaraldehyde was used and the signal-to-background ratio obtained was 2,400 or better. It was also shown that the europium signal was stable in daylight and at room temperature. Fluorescent europium chelate used in this experiment provides excellent contrast and long-term stability for the samples with glutaraldehyde fixation and permanent mounting. © 1994 Wiley-Liss, Inc.
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  • 100
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 459-467 
    ISSN: 1059-910X
    Keywords: Image analysis ; Self-assembly ; Persistence ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Deoxy-sickle hemoglobin (HbS) polymerizes in vivo into long helical fibers which fill the red cell and make it rigid. This impedes red cell passage through the capillaries and is responsible for the clinical manifestations of sickle cell disease. Images of individual and laterally associated HbS fibers were obtained by electron microscopy of frozen hydrated specimens. Each fiber possesses variable pitch, having from 6° to 12° rotation per unit cell. Laterally associated HbS fibers display systematic inter-fiber contacts in spite of their pitch variations, and exhibit better order than isolated fibers. This suggest that inter-fiber contacts can act to couple fibers mechanically and might therefore be a factor in rigidifying red cells in vivo. Fiber variability was attributed to local torsional variations with a standard deviation of 2.5°, but which are weakly coupled over a length of 2.25 unit cells. Variable pitch produces structural changes of as large as 5 Å azimuthally and 6 Å axially in HbS fiber unit cells. © 1994 Wiley-Liss, Inc.
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