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  • Articles  (56)
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  • Natural Sciences in General  (56)
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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 31 (1995), S. 234-247 
    ISSN: 1059-910X
    Keywords: Immunolocalisation ; Electron microscopy ; Ultrastructure ; hSP ; pS2 ; TFG α ; EGFR ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The trefoil peptides pS2 and human spasmolytic peptide are putative growth factors, particularly associated with mucus-producing cells of the gastrointestinal tract including those of the stomach. The receptor for transforming growth factor alpha (TGF α) takes its name from one of its alternative ligands, epidermal growth factor and is called the epidermal growth factor receptor. Although there is immunoreactive epidermal growth factor in the stomach, it is TGF α and the epidermal growth factor receptor that are abundant. Immunolabelling at electron microscope level allows for subcellular localisation of antigens; pS2 and human spasmolytic peptide co-localise to cytomembranes, including the Golgi apparatus, and thecae of surface/pit mucous cells. TGF α is abundant on the membranes of tubulovesicles of parietal cells and is also present in chief cells: in mucous producing cells it can be detected but not in association with mucous. The distribution of the epidermal growth factor receptor mimics that of TGF α but with preferential clustering on the basolateral membranes of gastric cells. The trefoil peptides are associated with healing and probably act, together with mucus, to protect the gastric mucosa and maintain a viable environment. TGF α, transduced via the epidermal growth factor receptor, inhibits gastric acid secretion, thus aids the trefoils in the maintenance of a gastric microenvironment conducive to healing after damage. TGF α, however, is also a potent mitogen; while this property plays a vital part in repairing mucosal defects, if this peptide or indeed its receptor are overexpressed, the result can be neoplasia. © 1995 Wiley-Liss, Inc.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 31 (1995), S. 215-225 
    ISSN: 1059-910X
    Keywords: Esophagus ; Epithelium ; Ciliated cells ; Esophagogastric junction ; Ultrastructure ; Human fetus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This article focusses on the structural development of human esophageal ciliated epithelium. A combination of transmission electron microscopic (TEM), scanning electron micro-scopic (SEM), radioautographic, and light microscopic (LM) analyses were carried out using intact fetal tissues between 8 and 20 weeks of gestation as well as cultured esophageal explants. Up to the age of 10 weeks, the stratified esophageal epithelium consisted of two longitudinal primary folds. The surface cells were undifferentiated and contained large glycogen aggregates. Between 11 and 16 weeks, the primary folds (now up to four) had developed secondary folds. The thickness of the epithelium drastically increased (123%) in concomittance with a differentiation of surface columnar ciliated cells. These highly specialized surface cells exhibited junctional complexes and well-developed organelles with numerous microvilli interspesed among the cilia. Transverse sections revealed the internal structure of the cilia with a consistent pattern of nine doublet microtubules surrounding a central pair of single microtubules. Freeze-fracture studies illustrated the presence of a ciliary necklace composed of 6 ring-like rows of intramembranous particles. They also revealed the structure of ciliary cell tight junctions consisting of up to nine anastomosing strands (P-face) or complementary grooves (E-face). Ultrastructural studies (LM, TEM, SEM) of the esophageal squamous epithelium obtained after 15 days of culture showed that the newly formed epithelium was similar to adult human epithelium. Finally LM and SEM observations established that the esophagogastric junction was not yet well delineated, consisting of a transitional area composed of a mixture of esophageal ciliated cells and gastric columnar mucous cells. © 1995 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 32 (1995), S. 129-147 
    ISSN: 1059-910X
    Keywords: Microscopy ; Ultrastructure ; Hyperplasia ; Hypertrophy ; Parathyroid hormones ; PTH synthesis ; Functional cycle ; Stereology ; Phosphate depletion ; Calcium depletion ; Uremia, Calcitriol [1,25(OH)2D3] ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ultrastructure of the rat parathyroid has been under study for more than 35 years, but controversies still exist, especially regarding structure-function relationships. The present review focuses on recent morphological parathyroid research on rats under normal conditions and in various states of disturbed calcium metabolism. To facilitate discussions on functional aspects, current biochemical data, particularly those dealing with the regulation of parathyroid hormone synthesis and release, are also considered. Our results from quantitative studies and from investigations employing serial sectioning form the basis for the discussions. A central issue is whether the parathyroid secretory cells undergo secretory cycles. Prompted by results obtained from improved fixation procedures and serial sectioning, we question the basis for the theory of secretory cycles. Since the rat parathyroid secretory cell is polar, a single section is not an appropriate sample for estimating functional activity and for comparing the structure and distribution of intracellular components of adjacent cells. The heterogeneity in ultrastructural appearance of intracellular vesicles calls for the use of specific markers in relating the structure of the vesicular compartment to intracellular processing of hormone. The importance of unbiased quantitative techniques is illustrated in discussions on cell number and size for estimating the response of the parathyroid gland to different functional states or disorders demanding changes in secretion of parathyroid hormone, e.g., hyper- and hypocalcemia. © 1995 Wiley-Liss, Inc.
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  • 4
    ISSN: 1059-910X
    Keywords: Sinus afferent pathway ; SP interneurons ; Double immunocytochemistry ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ultrastructure of substance P-containing nerve terminals synapsing on catecholamine neurons in the rat commissural subnucleus of the nucleus tractus solitarii (NTScom) was studied using a double immunocytochemical labeling technique. Although there were numerous tyrosine hydroxylase-immunoreactive (TH-I) somata present, substance P immunoreactive (SP-I) cell bodies were only occasionally found in the NTScom. At the light microscopic level, many SP-I terminals were seen closely associated with TH-I dendrites and somata. At the electron microscopic level, SP-I terminals synapsing on TH-I structures were also readily encountered. SP-I terminals contained small, clear, and predominantly spherical vesicles (32 ± 4 nm diameter), as well as large dense-cored vesicles approximately 100 nm in diameter. Postsynaptic TH-I dendritic profiles of various calibers and somata were encountered. These postsynaptic TH-I structures often showed postsynaptic densities. The morphological features of the SP-TH synapses in the present study, that is, the size of synaptic vesicles and the presence of postsynaptic densities, are quite different from those of central carotid sinus afferent synapses reported in our previous study [Chen et al. (1992), J. Neurocytol., 21:137-147]. Therefore, most of the SP terminals of the SP-TH synapses in the NTScom appear not to originate from the carotid sinus afferents. SP-I second-order neurons of the carotid sinus afferent pathway [Chen et al. (1991), J. Auton. Nerv. Syst., 33:97-98] may be one of the possible sources of such terminals. © 1994 Wiley-Liss, Inc.
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  • 5
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    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 169-176 
    ISSN: 1059-910X
    Keywords: Celiac ganglion ; Chromaffin cells ; Autonomic nervous system ; Ultrastructure ; Guinea pig ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Utilizing electron microscopic observation, several contacts between small, granule-containing cells (SGC) and postganglionic neurons (PGN) in the celiac ganglion of the guinea pig have been observed. A SGC in very close association with a PGN was seen to receive a distinct synaptic contact that contained many vesicles with dense cores. This contact was morphologically unlike cholinergic synapses previously reported on chromaffin cells. Because the SGC and PGN were clearly separated by a thin rim of satellite cell cytoplasm mutual to both cells, it is not known how or if the SGC would possibly exert a synaptic or paracrine effect on the PGN. Also, intraganglion SGC existed as large well-vascularized islands within the celiac ganglion. These intraganlion clusters sometimes contained more than 50 cells and perhaps could be considered to function as localized neuroendocrine components within the ganglion by secreting granule products into the nearby blood vessels for local or distant effects, although this certainly is not known. This work reports a unique synaptic ending upon a single-occurring SGC, which, in turn, closely approximates a ganglion neuron in a soma-somatic relationship. In addition, a very close association (but no actual contact) was observed between granule-containing processes, presumably emanating from the intraganglion clusters, and PGN. Whatever the function of ganglionic SGC may be, the exact relationship between SGC and PGN presumably would be of great interest and potential importance. © 1994 Wiley-Liss, Inc.
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  • 6
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 411-431 
    ISSN: 1059-910X
    Keywords: Corrosion casts ; LM ; SEM ; TEM ; Microvasculature ; Ultrastructure ; Absorption ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The aim of the present study was to provide a comprehensive morphological analysis of the porcine epididymis in view of the specific functions being performed in different regions of this organ. Blood supply and microvasculature of efferent ductules and epididymal duct were investigated by means of corrosion casts which were analysed macroscopically and by scanning electron microscopy. This revealed blood supply to the testis and epididymis to be closely related. The capillary pattern was typical for the efferent ductules, the caput, corpus, and distal cauda epididymidis, respectively. Corrosion casts were also used to visualize the course of the efferent ductules themselves. Tissue samples from different regions of the efferent ductules and epididymal duct were examined by light microscopy and both scanning and transmission electron microscopy, with special attention being payed to transitional areas. Morphological criteria allowed the distinction of three segments within the efferent ductules and of the initial segment, proximal caput, distal caput, corpus, proximal cauda, and distal cauda regions of the epididymal duct. Components of the endocytic apparatus of efferent ductule principal cells were identified by ferritin uptake. Ultrastructural evidence of absorption in the epididymal duct was particularly prominent in proximal and distal caput. Extensive cisternae of rough endoplasmic reticulum and a well-developed Golgi apparatus were indicative of active protein synthesis and secretion especially in the distal caput and corpus regions. However, assignment of various organelles in principal cells of the epididymal duct to either absorptive or secretory pathways still remains tentative. © 1994 Wiley-Liss, Inc.
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  • 7
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    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 145-164 
    ISSN: 1059-910X
    Keywords: Human ; Mouse ; Oocytes ; Maturation ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper reviews the process of peri-ovulatory oocyte maturation and the ultrastructural organization of the human egg and compares it with that of the mouse. The main thrust of the paper is on the human, since there are several reviews on the mouse. Both preovulatory and postovulatory events at fertilization, as well as some of the aberrant features of maturation are covered. Some changes induced by oocyte culture and cooling in the human are also included. The report attempts to focus on unique features of the human oocyte and shows a variety of ultrastructural differences between human and murine oocytes, which may well reflect differences in their physiology and biochemistry. Based on these differences and further observations on the process of fertilization of both species, particularly with respect to the inheritance of paternal centrioles, it is concluded that the mouse may not be a suitable modle for the development and refinement of current procedures in human assisted reproductive technology. © 1994 Wiley-Liss, Inc.
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  • 8
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    New York, NY [u.a.] : Wiley-Blackwell
    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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  • 9
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    New York, NY [u.a.] : Wiley-Blackwell
    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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  • 10
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 67-84 
    ISSN: 1059-910X
    Keywords: Intracellular labeling ; Horseradish peroxidase ; Lucifer Yellow ; Biocytin ; Neurobiotin ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have assessed the properties of three intracellular markers, horseradish peroxidase, biocytin/Neurobiotin, and Lucifer Yellow, and have compared their usefulness as neuronal markers for light and electron microscopic visualization. Neurons in the acute slice preparation of rat hippocampus were filled with one of these markers, and the marker was converted to an optical and electron-dense reaction product. Dimethylsulfoxide (DMSO) greatly facilitated penetration of recognition reagents while preserving membrane integrity. The markers were compared with respect to injection parameters, mobility and recognition, stability and visibility, and ultrastructural clarity. Horseradish peroxidase (HRP)-labeled neurons, recognized histochemically with diaminobenzedine (DAB), were easily visualized by the density of the DAB reaction product; however, the electron density was often so great as to obscure ultrastructural details. Biocytin (BC)-/Neurobiotin (NB)-labeled neurons were recognized by avidin-HRP, followed by histochemical localization of HRP with DAB. The optically dense reaction product gave complete visualization of the soma and processes at the light microscopic level. The electron density was homogeneously distributed throughout the cell, so that ultrastructural features were easily identified. Lucifer Yellow (LY), a fluorescent marker, was converted to an optical and electron-dense reaction product via immunocytochemical staining with a rabbit anti-LY antibody, followed by goat anti-rabbit IgG-HRP and DAB histochemical localization. Similar to BC/NB, the reaction product was evenly dispersed, providing good light microscopic and ultrastructural clarity. Under our experimental conditions, BC/NB and LY were superior markers that could be used routinely to label neurons, and give excellent visualization not only at the light but also at the electron microscopic level. © 1993 Wiley-Liss, Inc.
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  • 11
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    Microscopy Research and Technique 25 (1993), S. 148-168 
    ISSN: 1059-910X
    Keywords: Uterus ; Endometrium ; Decidua ; Collagen ; Ultrastructure ; Rodent ; Human ; Review ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Uterine aging is in part responsible for a decline in fecundity which begins in middle age in most mammals. Characteristics of uteri from a variety of animals in middle age and old age are reviewed and the factors which may be responsible for this decline discussed. These include age-related changes in the hypothalamus, pituitary, and ovaries; loss of number or function of steroid hormone receptors; morphological changes in the uterine epithelium; the accumulation of collagen fibrils in the uterine stroma; and loss or impairment of the decidual response. The ultra-structural morphology of uterine epithelial, stromal, and decidual tissue from 4 month old and 18 month old Fischer 344 rats is presented and compared. © 1993 Wiley-Liss, Inc.
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  • 12
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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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  • 13
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    Microscopy Research and Technique 20 (1992), S. 136-151 
    ISSN: 1059-910X
    Keywords: Pituitary ; Adenomas ; Tissue culture ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Morphologic studies of human adenohypophysial cells using immunocytochemistry and electron microscopy have characterized the hormone-producing cell types of the normal gland and pituitary adenomas. The classifications which have emerged allow more accurate clinicopathologic correlations than ever before, but have also raised new questions concerning cytogenesis, pathogenesis, and structure-function correlations. We report the results of studies which marry the conventional morphologic techniques of light microscopy, immunohistochemistry, electron microscopy, and ultrastructural immunocytology with functional analyses using tissue culture and radioimmunoassay of hormones released into culture media. The hormone secretory activity of nontumorous and adenomatous pituitary cells is correlated with their structural features; their secretory responses to several adenohypophysiotropic factors are compared with morphologic alterations which are characterized at the light and electron microscopic levels by morphometric analysis. These studies have shown that hypothalamic stimulating hormones increase hormone release by their target cells and alter the ultrastructural appearance of the affected cells by increasing organelles involved in hormone synthesis. Inhibitory drugs and adrenal and gonadal steroids are capable of suppressing hormone release by some tumors and also give rise to morphologic changes which correlate with the functional inhibition. Hormone release by clinically nonfunctioning adenomas has been characterized and the behavior of these tumor cells in vitro sheds some light on the reasons for lack of clinical symptomatology. The plurihormonal nature of several nontumorous and adenomatous pituitary cell types has been characterized in vitro. The results of these studies provide the basis for more accurate structure-function correlations which can be used to study the hormonal milieu in vivo, to predict the role of pathogenetic factors in pituitary tumorigenesis, and to assess the therapeutic value of stimulating or inhibiting hormones and drugs.
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  • 14
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    Microscopy Research and Technique 22 (1992), S. 103-125 
    ISSN: 1059-910X
    Keywords: Cell lineage analysis ; Mammalian embryo ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ultrastructural studies have contributed significantly to our understanding of cell lineage differentiation in the mammalian pre-implantation embryo. Such studies have documented, and continue to document, morphological, biochemical, and physiological characteristics of the cell lineages established during the pre-implantation period in eutherian embryos, principally that of the mouse. This review evaluates these contributions and identifies areas of study in which ultrastructural analysis is most likely to have an important role in the future. © 1992 Wiley-Liss, Inc.
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  • 15
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    Microscopy Research and Technique 23 (1992), S. 62-75 
    ISSN: 1059-910X
    Keywords: Olfactory receptor cell ; Supporting cell ; Ultrastructure ; Lipofuscin granules ; Golgi apparatus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The present study uses mainly scanning electron microscopy to demonstrate the three-dimensional internal cell structures of rat olfactory epithelial cells. The aldehyde-prefixed osmium-DMSO-osmium (AODO) method devised by Tanaka and Mitsushima (1984) was applied to the present study to disclose intracellular structures such as endoplasmic reticulum, mitochondria, Golgi apparatus, and lysosomes. The spatial distribution pattern of these structures in olfactory and supporting cells is discussed, paying special attention to the formation of lipofuscin-like granules present in aged rats. © 1992 Wiley-Liss, Inc.
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  • 16
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    Microscopy Research and Technique 23 (1992), S. 22-27 
    ISSN: 1059-910X
    Keywords: Plasticity ; Retrograde degeneration ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We used scanning (SEM) and transmission (TEM) electron microscopy to examine ultrastructural changes in the olfactory epithelium (OE) of rainbow trout following unilateral olfactory nerve section. Both ciliated receptor cells (CRC) and microvillar receptor cells (MRC) degenerated and subsequently differentiated from unidentified precursor cells. The following changes took place in fish that were held at 10°C at the stated period following olfactory nerve section: on day 7, MRC and CRC contained intracellular vacuoles; on day 12, the olfactory knobs appeared disrupted; by day 26, olfactory receptor cells were absent from the OE; on day 42, there were receptor cell bodies and a few CRC with short cilia at the apical surface; and opn day 55, a small number of both CRC and MRC had differentiated. By day 76, both CRC and MRC repopulated the OE. Degenerative changes in the cytoplasm of the sustentacular cells (SC) and ciliated nonsensory cells (CNC) were observed in the first 26 days following olfactory nerve section, but these cells remained intact throughout the experiment. The degeneration and subsequent differentiation of CRC and MRC supports and extends previous observations that both cells types are olfactory receptor neurons with axons that extend along the olfactory nerve to the olfactory bulb. © 1992 Wiley-Liss, Inc.
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  • 17
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    Microscopy Research and Technique 23 (1992), S. 28-48 
    ISSN: 1059-910X
    Keywords: Nose ; Olfaction ; Ultrastructure ; Toxicology ; Smell ; Sensory ; Fish ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes four investigations of the olfactory mucosa of the brown trout: 1) the ultrastructure of the olfactory mucosa as revealed by scanning (SEM), conventional transmission (TEM), and high voltage (HVEM) electron microscopy; 2) light and electron-microscopic investigations of retrograde transport of the tracer macromolecule horseradish peroxidase (HRP) when applied to the cut olfactory nerve; 3) SEM and TEM investigations of the effects of olfactory nerve transection on cell populations within the olfactory epithelium; and 4) ultrastructural investigations of reversible degeneration of olfactery receptors caused by elevated copper concentrations. The trout lofactory epithelium contains five cell types: ciliated epithelial cells, ciliated olfactory receptor cells, microvillar olfactory receptor cells, supporting cells, and basal cells. The ciliated and microvillar olfactory receptor cells and a small number of basal cells are backfilled by HRP when the tracer is applied to the cut olfactory nerve. When the olfactory nerve is cut, both ciliated and microvillar olfactory receptor cells degenerate within 2 days and are morphologically intact again within 8 days. When wild trout are taken from their native stream and placed in tanks with elevated copper concentrations, ciliated and microvillar cells degenerate. Replacement of these trout into their stream of origin is followed by morphologic restoration of both types of olfactory receptor cells. Ciliated and microvillar receptor cells are primary sensory bipolar neurons whose dendrites make contact with the environment; their axons travel directly to the brain. Consequently, substances can be transported directly from the environment into the brain via these “naked neurons.” Since fish cannot escape from the water in which they swim, and since that water may occasionally contain brain-toxic substances, the ability to close off - and later reopen - this anatomic gateway to the brain would confer a tremendous selective advantage upon animals that evolved the “brain-sparing” capacity to do so. Consequently, the unique regenerative powers of vertebrate olfactory receptor neurons may have their evolutionary origin in fishes. © 1992 Wiley-Liss, Inc.
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  • 18
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    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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  • 19
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    Microscopy Research and Technique 23 (1992), S. 1-21 
    ISSN: 1059-910X
    Keywords: Cilia ; Microvilli ; Ultrastructure ; Electron microscopy ; Evolution ; Cladistics ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this paper, the evolutionary origin of the vomeronasal system as a discrete sensory system separate from olfaction is examined. The presence of a discrete vomeronasal system appears to be a derived character in tetrapods, and its presence in larval amphibians indicates that the system did not arise as a terrestrial adaptation. The vomeronasal system has been lost independently in several taxa, including crocodilians, some bats, cetaceans, and some primates. The presence of microvillar receptor cells in the vomeronasal epithelium appears to be the ancestral condition for tetrapods, and alternative hypotheses concerning the ancestral condition for receptor cell types in the vertebrate olfactory epithelium are discussed. Finally, the possibility that the vomeronasal system is present in some fishes in a form that has not been recognized is discussed in relation to the phylogenetic distribution of receptor cell types in vertebrates. © 1992 Wiley-Liss, Inc.
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  • 20
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    Microscopy Research and Technique 22 (1992), S. 307-324 
    ISSN: 1059-910X
    Keywords: Sensory ; Chemosensory ; Ultrastructure ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Among gastropod molluscs the chemical senses are most important for location of distant objects. They are used in food finding, locating mates, avoiding predators, trail following, and homing. Chemoreceptors are commonly associated with the oral area, the tentacles, and the osphradium, which lies in the mantle cavity.Most chemosensory neurons are primary sensory neurons, although secondary sensory cells have been reported in the osphradium of some prosobranch gastropods. Most chemosensory organs contain sensory cells with ciliated sensory endings that are in contact with the external environment. Some sensory endings have only microvilli or have no surface elaborations. Cilia on sensory endings are commonly of the conventional type, but some species have modified cilia; some lack rootlets, some have an abnormal microtubular content, and some have paddle-shaped endings. The perikarya of sensory neurons may be within the sensory epithelium, below it, or in ganglia near the sensory surface. In some groups of gastropods there are peripheral ganglia in the olfactory pathway; in others chemosensory axons appear to pass directly to the CNS.Olfactory epithelia of terrestrial pulmonates have modified brush borders with long branching plasmatic processes and a spongy layer of cytoplasmic tubules which extend from the epithelial cells. Sensory endings of the olfactory receptors are entirely within this spongy layer. Aquatic pulmonates may have a similar spongy layer in their olfactory epithelia, but the cilia of sensory endings, as well as motile cilia of epithelial cells, extend well beyond the spongy layer. © 1992 Wiley-Liss, Inc.
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  • 21
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    Microscopy Research and Technique 21 (1992), S. 53-58 
    ISSN: 1059-910X
    Keywords: Naphthalene ; Dislocations ; Replicas ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: It is interesting to apply the method of etch figures to the study of organic molecular crystal defects, by observing the etch pits as soon as they are produced. We have set up a method to determine the geometrical forms of such small etch pits, observed on pre-shadowed replicas of naphthalene crystal surfaces. The described experimental procedure was designed to avoid artefacts due to vacuum sublimation and moisture traces on the replicated surface. Stereoscopic observation makes interpretation possible. The 3-D morphology and size of etch figures smaller than 1 μm can be determined.
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  • 22
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    Microscopy Research and Technique 21 (1992), S. 136-157 
    ISSN: 1059-910X
    Keywords: Pinealocytes ; Ultrastructure ; Secretion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The primary aim of this review is to present the current state of knowledge of the ultrastructure of the mammalian pineal gland, with emphasis on its functional aspects. Basic ultrastructural features of the mammalian pinealocytes are presented with special attention paid to ultrastructural aspects of pineal secretion.
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  • 23
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    Microscopy Research and Technique 23 (1992), S. 128-141 
    ISSN: 1059-910X
    Keywords: Olfactory epithelium ; Vomeronasal epithelium ; Endocytosis ; Ultrastructure ; Olfactory pigment ; Mouse ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Mammalian olfactory neurons possess a well-developed system of endocytic vesicles, endosomes, and lysosomes in their dendrites and perikarya. Vomeronasal neurons are similar and also contain much perikaryal agranular endoplasmic reticulum (AER). Olfactory supporting cells contain endocytic vesicles and endosomes associated closely with abundant fenestrated AER, and vesicles and numerous large dense vacuoles are present basally. Vomeronasal supporting cells have little AER, and few dense vacuoles occur in their bases. In olfactory neurons, ultrastructural tracers (0.08% horseradish peroxidase, thorium dioxide, ferritin) are endocytosed by olfactory receptor endings and transported to the cell body, where their movement is halted in lysosomes. Higher concentrations (1%) of horseradish peroxidase penetrate olfactory receptor plasma membranes and intercellular junctions. In olfactory supporting cells, endocytosed tracers pass through endosomes to accumulate in dense basal vacuoles. These observations indicate that olfactory sensory membranes are rapidly cycled and that endocytosed materials are trapped within the epithelium. It is proposed that in the olfactory epithelium, endocytosis presents redundant odorants to the enzymes of the supporting cell AER to prevent their accumulation, whereas in the vomeronasal epithelium the receptor cells carry out this activity. © 1992 Wiley-Liss, Inc.
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  • 24
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    Microscopy Research and Technique 23 (1992), S. 142-156 
    ISSN: 1059-910X
    Keywords: Cytoskeleton ; Ultrastructure ; Microtubules ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The olfactory neuron is specialized along its length into highly determined morphological regions. These regions include the dendritic cilia, dendritic vesicle, dendritic shaft proper, perikaryon, axon, zone of transition where the axon widens as it approaches its termination, and the axon terminal. Except for the zone of transition and the terminal, characteristic populations of microtubules occur in these compartments. In the olfactory vesicle, three discrete microtubule organizing centers (MTOCs) nucleate microtubules: the basal body, the lateral foot associated with the body, and dense masses of nearby material. Little is known about MTOCs elsewhere in the neuron, although the polarity of the axonal microtubules indicate that they originate at or near the perikaryon. An attempt is made to summarize what is known of the origin, structure, distribution, and function of microtubules in vertebrate olfactory neurons, which are useful model systems in which to study microtubules. Information about olfactory neuron microtubules may be applicable to neurons in general (e.g., the discovery that axons contain microtubules of uniform polarity was first made in the olfactory neuron) or to microtubules in other eukaryotic cells. © 1992 Wiley-Liss, Inc.
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  • 25
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    Microscopy Research and Technique 23 (1992), S. 225-229 
    ISSN: 1059-910X
    Keywords: Scanning electron microscopy ; Ultrastructure ; Zona pellucida ; Mucus ; 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 zona pellucida (ZP) covering the oocyte and of the mucus covering the surface of the intestinal villi was investigated by using a new method employing ruthenium red (RR), saponin, and osmium-thiocarbohydrazide impregnation.The glycoproteic matrices both appeared constituted by thin filaments (ranging from 22 to 50 nm in thickness) anastomosed to form a very fine network.RR prevented the dissolution and/or alteration of glycoproteins and polyanionic carbohydrates induced by acqueous fixatives. Saponin was a detergent of the soluble proteins. Osmium-thiocarbohydrazide preserved the glycoproteic matrix filaments from the mechanical stress induced by dehydration and critical point drying and reduced filaments packing and shrinkage. The technical improvement was demonstrated by the following results: 1) a regular arrangement of the filaments network; 2) a thickness of mucus filaments smaller than that obtained with other methods of preparation; 3) a homogeneous thickness of ZP filaments.This method allowed a very detailed study of the fine structural organization of the ZP and intestinal mucus. Therefore, this technique can be useful for a better evaluation of the morphodynamic of these and other glycoproteic matrices. © 1992 Wiley-Liss, Inc.
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  • 26
    ISSN: 1059-910X
    Keywords: Ultrastructure ; Immunocytochemistry ; Chromatin structure ; Nuclear proteins ; Testis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have followed the fine structural distribution of two nucleosomal core histones, H2B and H3, and of protamines in the course of mouse spermiogenesis by means of specific antibodies and ultrastructural immunocytochemistry.Our results demonstrate that the nuclear labeling density of histone H2B decreases during steps 6-8 and then increases again in step 9-10 spermatids, while the labeling for histone H3 is constant throughout this period. In step 12 spermatids, the anti-H2B antibody labels mainly the central area of the nucleus. The first signs of protamine labeling are present in step 12 spermatids, where the gold grains can be found over the periphery of the nucleus. Later on, protamine labeling constantly increases and, by the end of spermiogenesis, the whole nucleus is labeled.We suggest that the morphological and structural differences between the central area and the periphery of mouse spermatids are, at least partly, due to a difference in the protein moiety associated with DNA. The central area, which is peculiar to the mouse and has been previously considered as a focus of chromatin condensation, represents, however, the last nuclear region containing histones and consequently the last area where the substitution of histones by protamines takes place.
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  • 27
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    Journal of Electron Microscopy Technique 18 (1991), S. 74-81 
    ISSN: 0741-0581
    Keywords: Confocal scanning laser microscopy ; Connective tissue ; Elastic tissue ; Eye ; Microvasculature ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The connections between elastic tissue and microvessels (arterioles, capillaries, and venules) in the rabbit eye were examined by light and electron microscopy. In particular, confocal scanning laser microscopy of tissue stained with orcein and examined by fluorescence using a rhodamine filter was correlated with electron microscopic observations. The goal was an analysis of the way in which elastic tissue of the uvea (i.e., choroid, ciliary body, and iris) and the optic nerve of the eye connect to the microvessels in these structures. Confocal microscopy revealed these connections advantageously and showed how they link the elastic tissue meshwork in the perivascular tissue spaces with the wall of the blood vessels. Electron microscopy showed that the connections consist of bundles of 10-12 nm diameter microfilaments that insert into vascular basement membranes. These connections may contribute to the vascular response to changes in blood pressure or intraocular pressure in the eye.
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  • 28
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    Journal of Electron Microscopy Technique 19 (1991), S. 172-188 
    ISSN: 0741-0581
    Keywords: Testis ; Morphogenesis ; Cytodifferentiation ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The fetal testis is not merely a precursor of the adult organ: it is indeed an endocrine gland whose function is the masculinization of the fetus. It differs physiologically and morphologically from the adult testis. In this paper, the first stages of testicular differentiation in the rat are described, with special emphasis on the ultrastructural aspects. At the stage of 13.5 days after fertilization, the first Sertoli cells differentiate; they are characterized by a voluminous and little electron dense cytoplasm, a well-developed RER formed by vesicles and short cisternae filled with a flocculent material. Progressively, they polarize and adhere to one another by adherens-like junctions and cytoplasmic interdigitations to form the differentiating seminiferous cords. In the basal part of the Sertoli cells, a mat of microfilaments differentiates under the plasmalemma, while cytoplasmic blebs protruding in the extracellular space tend to disappear. A continuous basal lamina delineating the seminiferous cords begins to appear on day 14.5 and becomes widespread on day 15.5. These observations, when compared with other data from the literature, emphasize the fact that the differentiation of the Sertoli cells is the first morphological event during testicular differentiation. A possible role of the Sertoli cells in the subsequent organogenesis of the testis is suggested.
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  • 29
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    Journal of Electron Microscopy Technique 17 (1991), S. 412-436 
    ISSN: 0741-0581
    Keywords: Human sperm ; Ultrastructure ; Pathology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In this article, the major contributions of electron microscopy to the present understanding of the physiology and pathophysiology of the human spermatozoon are reviewed. The ultrastructural organization of sperm organelles playing a significant role for cell function and, therefore, for the reproductive process is described. Also, the major abnormalities and defects of the various organellar systems and how they impair the reproductive function and/or the viability of the cell are reviewed.
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  • 30
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    Journal of Electron Microscopy Technique 19 (1991), S. 276-290 
    ISSN: 0741-0581
    Keywords: Cryofixation ; Endothelium ; Blood Vessels ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cryofixation refers to the immobilization of tissue components by the rapid removal of heat from the specimen, so that the structure is interred and stabilized in a natural embedding medium, namely, frozen (amorphous or microcrystalline) tissue water. Cryofixation is now often used as a complement to the more traditional fixation methods, especially when the cell structure is delicate or dynamic and may be inaccurately preserved by the slow selective action of chemical fixatives. Vascular endothelial cells are specialized for transcellular transport and for the regulation of blood flow and composition. The dynamic and labile subcellular organization of these cells, presumably reflecting these functional specializations, makes them ideal candidates for cryofixation.Several different types of endothelial cells were directly frozen at temperatures below 20 degrees Kelvin by pressing them against a liquid-helium-cooled block. These samples were subsequently processed for structural analysis by freeze-substitution. Detailed rationales, designs, and protocols are described for both freezing and freeze-substitution.Electron micrographs of cryofixed arterial and venous capillaries (rete mirabile of the American eel), iliac vein (rabbit), and cultured endothelium from the iliac vein (human) reveal that the organization of the characteristic intracellular membrane system of endothelial vesicles is qualitatively similar to that seen in chemically fixed endothelium, especially with regard to the interconnection of clusters of individual vesicles to form elaborate networks. The luminal and abluminal networks are not in communication, at least not in static images. Quantitatively, however, most directly frozen endothelial cells have far fewer vesicular profiles than comparable glutaraldehydefixed cells. The differences can be explained by presuming that the rapid action of cryofixation (approximately 1 msec) gives a more accurate picture of the vesicular network because it captures the transient structure of labile or dynamic membranes.
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  • 31
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    Journal of Electron Microscopy Technique 19 (1991), S. 107-117 
    ISSN: 0741-0581
    Keywords: Plant ; Leaf ; Root ; Tissue ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cryofixation and freeze substitution methods were developed for ultrastructural studies of cells in complex plant tissues. Leaf tissues and root tips of tobacco (Nicotiana tabacum L. var. Maryland Mammoth) were frozen with a RMC MF7200 propane jet freezer and freeze substituted sequentially with tannic acid and osmium tetroxide/uranyl acetate in acetone. High quality preservation was consistently obtained for epidermal and phloem cells of the leaf, and epidermal, cortical, meristematic, and cap cells of the root tip. Leaf mesophyll cells were also often well frozen. Organelles, including nuclei, endoplasmic reticulum, mitochondria, Golgi bodies, and plastids, showed excellent structural integrity and contrast. Most notable is the superior preservation of the cytoskeleton. Our results demonstrate that the propane jet freezer can be used routinely for high quality cryofixation of higher plant cells in certain complex tissues. This could have important implications for the use of cryofixation approach in a wide range of research in plant biology.
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  • 32
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    Journal of Electron Microscopy Technique 14 (1990), S. 106-125 
    ISSN: 0741-0581
    Keywords: Aging ; Ultrastructure ; Liver volume ; Lysosomes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Aging is accompanied by a myriad of changes in cell structure, function, and composition. The fact that much of the information concerning age-related alterations in cellular morphology is qualitative precludes meaningful correlations with biochemical changes in order to enhance data interpretation. The mammalian liver has been subjected to both qualitative and quantitative evaluations of hepatocyte structure as a function of aging, i.e., development, maturation, and senescence. Although these data are characterized by considerable variability and, in some instances, blatant contradictions, there exists sufficient agreement in several parameters to permit a consensus in the inbred rat model. Certainly the volume of individual hepatocytes increases with age, and many of the organelle compartments reflect this change. While old rats exhibit a high incidence of polyploidy, there is no definitive evidence to demonstrate a concomitant increase in the binuclear hepatocyte index. Several specific hepatocellular organelles undergo changes in their relative volume or surface area that appear to correlate with functional alterations. The volume density of the lysosomal compartment enlarges significantly during senescence and is accompanied by increased activities of several constituent hydrolases. The hepatic concentration of smooth-surfaced endoplasmic reticulum declines markedly with aging, as does the yield of liver microsomes and the activities of several microsomal enzymes, e.g., mono-oxygenases and glucose-6-phosphatase. However, the responses of the majority of hepatocyte organelles to aging is varied and inconsistent based on the limited data currently available.
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  • 33
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    Journal of Electron Microscopy Technique 14 (1990), S. 179-207 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Liver disease ; Hepatitis ; Alcoholic injury ; Storage diseases ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Hepatocytes respond to injury by a few basic pathological reactions that are reflected in cell death, different types of degeneration, regeneration, or tumorous transformation. At the ultrastructural level, alterations of cell organelles can be observed in different combinations as a result of the injury, depending on the etiological agent(s) or pathological conditions developed. Nuclear bodies, dilation and fragmentation of rough endoplasmic reticulum (rer), swelling of mitochondria, and an increased number of lysosomes occur during acute viral hepatitis. The core and surface components of the hepatitis B virus can be localized in the liver cells in chronic hepatitis and in carriers. Close contact of hepatocytic and lymphocytic cell membranes were observed in chronic active hepatitis. Hepatocytes surrounded by an increased amount of collagen fibers are characteristic of cirrhosis. Loosely arranged, fine fibrils or condensed forms of Mallory bodies are pathognomic for alcoholic injury. A wide spectrum of alterations are noted after drug treatment: the proliferation of smooth endoplasmic reticulum (ser) as an adaptive phenomenon, focal or complete necrosis of the cell, inflammation, and the like. The fine structural analysis of hepatocytic inclusions in storage diseases has a differential diagnostic value. The storage of copper and other elements can be measured by x-ray microanalysis. The study of the hepatocytic differentiation in liver tumors is highly important in establishing the diagnosis and in proving the hepatocytic origin of the tumor.
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  • 34
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    Journal of Electron Microscopy Technique 16 (1990), S. 93-114 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Biochemistry ; Maturation ; Comparative ; Oocytes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This review of the anatomical, histological, biochemical, and molecular biological literature on echinoderm oogenesis includes the entire developmental history of oocytes; from their inception to the time they become ova. This is done from a comparative perspective, with reference to members of the five extant echinoderm classes; crinoids, holothurians, asteroids, ophiuroids, and echinoids. I describe the anatomy and fine structure of the echinoderm ovary, with emphasis on both the cellular relationships of the germ line cells to the somatic cells of the inner epithelium, and on the neuromuscular systems. I review the literature on the growth of oogonia into fully formed oocytes, including the process of vitellogenesis, presenting an ultrastructural analysis of the organelles and extracellular structures found in fully formed echinoderm oocytes. Echinoderm oocyte maturation is reviewed and a description of the ultrastructural, biochemical and molecular biological changes thought to occur during this process is presented. Finally, I discuss oocyte ovulation, the severing of cellular connections between the oocyte and its surrounding somatic epithelial cells.
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  • 35
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    Journal of Electron Microscopy Technique 16 (1990), S. 202-234 
    ISSN: 0741-0581
    Keywords: Xenopus ; Meiosis ; Ultrastructure ; Intracellular signals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Amphibian oocytes, arrested in prophase I, are stimulated to progress to metaphase II by progesterone. This process is referred to as meiotic maturation and transforms the oocyte, which cannot support the early events of embryogenesis, into the egg, which can. Meiotic maturation entails global reorganization of cell ultrastructure: In the cell cortex, the plasma membrane flattens and the cortical granules undergo redistribution. In the cell periphery, the annulate lamellae disassemble and the mitochondria become dispersed. In the cell interior, the germinal vesicle becomes disassembled and the meiotic spindles form. Marked changes in the cytoskeleton and mRNA distribution also occur throughout the cell. All of these events are temporally correlated with intracellular signalling events: Fluctuations in cAMP levels, changes in pH, phosphorylation and dephosphorylation, and ion flux changes. Evidence suggests that specific intracellular signals are responsible for specific reorganizations of ultrastructure and mRNA distribution.
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  • 36
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    Journal of Electron Microscopy Technique 15 (1990), S. 20-33 
    ISSN: 0741-0581
    Keywords: Catecholamines ; Ultrastructure ; Synaptic terminals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The monoamines dopamine, noradrenaline, adrenaline, and serotonin as well as the diamine histamine have a widespread distribution in the central nervous system within synaptic terminals and nonsynaptic varicosities. In certain regions of the central nervous system the monoamines are contained in varicosities that have no synaptic specialization associated with them, suggesting a possible neuromodulatory role for some of the monoamines. The majority of monoamine labelled structures are synaptic terminals which are characterized by the presence of small, clear vesicles (40-60 nm) and large, granular vesicles (70-120 nm) within the terminal. A third population of vesicles - small, granular vesicles - which are visible only after histochemical staining, are probably the equivalent of the small, clear vesicles present after either autoradiographic or immunohistochemical labelling. Most monoamine containing terminals contact dendrites and dendritic spines and, less frequently, neuronal somata and other axons. Both asymmetrical and symmetrical membrane specializations are associated with monoaminergic terminals; however, asymmetrical contacts are the most frequent type found. These ultrastructural results indicate that monoamine containing terminals and varicosities in general share many common morphological features, but still have diverse functions.
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  • 37
    ISSN: 0741-0581
    Keywords: Lectins ; Ultrastructure ; Visual system ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Phaseolus vulgaris leucoagglutinin (PHA-L) is a plant lectin that is anterogradely transported by neurons in the central nervous system. PHA-L is selectively taken up by cells at iontophoretic injection sites and, when immunohistochemically demonstrated, labels individual neurons completely, including their dendrites, axons, and terminal boutons. PHA-L is generally not taken up by fibers passing through the injection site and, because it produces a Golgi-like staining of even very fine axons over long distances, it is sometimes possible to light microscopically reconstruct individual neurons and their entire axon terminal arbors. When prepared for electron microscopy, the PHA-L-labeled terminals are densely and completely stained, allowing their synaptic relationships to be defined. These properties make PHA-L advantageous for studying the patterns of projection and the modes of termination of select groups of neurons in their target nuclei.We used PHA-L to study the extraretinal innervation of the cat's dorsal lateral geniculate nucleus, a thalamic visual center. Although much is known about the retinal contribution to geniculate synaptic circuitry, relatively little is known about other sources of innervation, even though these provide the majority of synaptic terminals in the nucleus (Guillery: Z. Zellforsch., 96:1-38, 39-48, 1969; Wilson et al.: Proc. R. Soc. Lond. [Biol.], 221:441-436, 1984). We used both light and electron microscopy to describe synaptic circuitry from three extraretinal sources of projections to the lateral geniculate nucleus: the visual cortex, the perigeniculate nucleus, and the parabrachial region of the brainstem. Cortical terminals labeled with PHA-L were small and formed asymmetrical synaptic contacts onto small-caliber dendrites of geniculate neurons. Peri-geniculate terminals formed symmetrical synaptic contacts primarily onto small-caliber dendrites, but some synapses were also formed onto the proximal, retinorecipient portions of geniculate dendrites. Parabrachial terminals synaptically contacted the retinorecipient portions of dendritic appendages and shafts, small-caliber dendrites, and the specialized dendritic (F2) terminals of geniculate interneurons. The symmetry of the parabrachial synaptic contacts was variable and was related to the postsynaptic target. Contacts onto dendritic appendages were asymmetrical while those onto dendritic shafts and F2 terminals were symmetrical. Our data suggest that in unlabeled material these brainstem terminals would be difficult to distinguish from cortical or perigeniculate profiles. The positioning of the parabrachial input onto the retinorecipient portions of geniculate dendrites indicates that this projection is well situated to control primary retinal transmission through the nucleus, while the location of most cortical and perigeniculate innervations implicates them in secondary feedback interactions or other aspects of geniculate function.
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    Journal of Electron Microscopy Technique 16 (1990), S. 2-14 
    ISSN: 0741-0581
    Keywords: Intestine ; Epithelium ; Mucous cells ; Nucleolus ; Cell kinetics ; Ultrastructure ; Protein synthesis ; RNA synthesis ; Stem cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This article is a summary of our work of several years on the renewal of the intestinal epithelium. A combination of ultrastructural, radioautographic, and light microscopic analyses was carried out using normal tissue and tissue affected by inhibitors of RNA and protein synthesis. Measuring protein synthesis by 3H-leucine radioautography showed that the life span of the columnar (absorptive) cells in the rat small intestine was divisible into two main phases: differentiation (from stem to functional cell) and maturation (from functional to extruding cell), each phase and its subdivisions being well defined morphologically. Differentiation involved a linear rise in the rate of protein synthesis per cell and showed at the same time heterochromatinization and silencing of RNA transcription. Data from various experiments indicated that the cells functioned from stored information (RNA), part of which came from the nucleolus, which underwent marked and characteristic ultrastructural changes. Although transcription from rDNA ceased, the nucleolus released its ribosomal material, which added to the existing protein synthesis, presumably by recruiting excess stored mRNAs. Maturation involved a nearly linear decrease of the rate of protein synthesis per cell to a characteristic low value at which extrusion took place. A gradual exhaustion of the stored RNA was indicated to be the key factor in this decrease. Ultrastructurally, maturation was associated with a gradually increasing vesiculation of rER and Golgi. The results thus imply a regulatory role of cellular protein synthesis level in renewal. This would be an epigenetic response after the genes are silenced. The nucleolus seems to play a central role in this process, and this in turn is reflected in its characteristic ultrastructural changes. The work also included new observations on the epithelium of the rat ascending colon describing a hitherto unrecognized deep crypt mucus-secretory (“DCS”) cell which is a nongoblet mature cell type apparently arising from midcrypt mitoses. In between the DCS cells, occasional slender columnar cells were seen which displayed the ultrastructural features of stem cells. These were probably reserve stem cells. We also observed nongoblet deep crypt mucous cells in the human right colon although fewer in number than in the rat. Nucleolar regulation and the presence of reserve stem cells represent new dimensions in our understanding of renewal. Electron microscopy is an essential tool in this investigation.
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  • 39
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    Journal of Electron Microscopy Technique 15 (1990), S. 2-19 
    ISSN: 0741-0581
    Keywords: Acetylcholine ; Cholinergic fibers ; Electron microscopy ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cholinergic synapses can be identified in immunocytochemical preparations by the use of monoclonal antibodies and specific antisera to choline acetyltransferase (ChAT), the synthesizing enzyme for acetylcholine (ACh) and a specific marker for cholinergic neurons. Electron microscopic studies demonstrate that the fibers and varicosities observed in light microscopic preparations of many brain regions are small-diameter unmyelinated axons and vesicle-containing boutons. The labeled boutons generally contain clear vesicles and one or more mitochondrial profiles. Many of these boutons form synaptic contacts, and the synapses are frequently of the symmetric type, displaying thin postsynaptic densities and relatively short contact zones. However, ChAT-labeled synapses with asymmetric junctions are also observed, and their frequency varies among different brain regions. Unlabeled dendritic shafts are the most common postsynaptic elements in virtually all regions examined although other neuronal elements, including dendritic spines and neuronal somata, also receive some cholinergic innervation. ChAT-labeled boutons form synaptic contacts with several different types of unlabeled neurons within the same brain region. Such findings are consistent with a generally diffuse pattern of cholinergic innervation in many parts of the central nervous system. Despite many similarities in the characteristics of ChAT-labeled synapses, there appears to be some heterogeneity in the cholinergic innervation within as well as among brain regions. Differences are observed in the sizes of ChAT-immunoreactive boutons, the types of synaptic contacts, and the predominant postsynaptic elements. Thus, the cholinergic system presents interesting challenges for future studies of the morphological organization and related function of cholinergic synapses.
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  • 40
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    Journal of Electron Microscopy Technique 15 (1990), S. 144-154 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Spiral ganglion ; Ontogenesis ; Myelination ; TII neurons ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The development of the spiral ganglion in the cat, the rat, and the mouse was studied by electron microscopy, from fetal stages in the cat and from birth in the rodent. In the earliest stages, a single population of ganglion cells is present. Immature spiral ganglion neurons possess small perisomatic processes that seem to disappear with development, before the myelination ganglion cells are surrounded by one or two layers of Schwann cell processes. With maturation, the Schwann process increases in number around the perikaryon and its processes, which leads to the onset of myelination. The onset of myelination of the cell body processes is asynchronous. The perikaryon may be delayed in myelination by several days. Moreover, ganglion neurons from a given region of the cochlea do not myelinate simultaneously. The differentiation of two types of fibers in the intraganglionic spiral bundle and the first appearance of TII neurons occurs around birth in the cat and a few days after birth for the rat and the mouse. The distinction of TII cells is possible due to characteristic accumulation of neurofilamentous structures in the cytoplasm.
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  • 41
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    Journal of Electron Microscopy Technique 16 (1990), S. 257-280 
    ISSN: 0741-0581
    Keywords: Oocyte ; Maturation ; Ultrastructure ; Gap junction ; Cortex ; Mammal ; Granulosa ; Actin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Immature mammalian oocytes reside in ovarian follicles with junctionally coupled granulosa cells. When released from a currently undefined meiotic arresting influence, these oocytes resume meiosis to progress from late diplotene (germinal vesicle stage) through the first meiotic division to metaphase II. Oocytes remain at metaphase II until fertilization activates them to complete meiosis. This review summarizes ultrastructural events that occur during meiotic maturation in mammals. Developmental correlates that promise a clearer understanding of regulatory mechanisms operating to control maturation are emphasized. By use of TEM of thin sections, freeze-fracture analysis, and replicated oocyte cortical patches, we demonstrate stage-specific changes in the oocyte nucleus, reorganization of cytoplasmic organelles, correlations between oocyte maturational commitment and the junctional integrity of associated granulosa cells, and definition of the components comprising the oocyte cortical cytoplasm.
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  • 42
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    Journal of Electron Microscopy Technique 12 (1989), S. 24-28 
    ISSN: 0741-0581
    Keywords: Tissue culture ; Heart muscle cells ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A technique for performing light, scanning, and transverse transmission electron microscopy on cultured cells grown within a single tissue culture flask is described. Permanent light microscopy slides are obtained by removing selected portions of the plastic tissue culture vessel and mounting them on glass slides with an aqueous mounting solution. The images obtained from these slides are superior to viewing through the bottom of the flask with an inverted stage microscope. For scanning electron microscopy, selected areas are also cut from the remainder of the vessel and prepared for viewing. The final portion of the culture container is transferred and attached to a new tissue culture vessel and prepared for transmission electron microscopy using alcohol instead of acetone and propylene oxide during dehydration, infiltration, and embedding.
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  • 43
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    Journal of Electron Microscopy Technique 12 (1989), S. 323-330 
    ISSN: 0741-0581
    Keywords: Chromaffin cells ; Paraganglia ; Paraaortic organs ; Mitosis ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Mitotic activity often has been reported in embryonic and fetal sympathetic neuroblasts, principal sympathoblasts, and primitive sympathetic cells in various species at different stages of development. Postnatal adrenal medullary cells also are known to undergo mitosis, but such dividing capabilities rarely have been observed in the true postnatal extraadrenal chromaffin system. Although few in number, this work nevertheless has clearly identified such cells in varying stages of the mitotic cycle in the young dog, Syrian hamster, mouse, rabbit, and rat. The dividing cells were noted in paraaortic chromaffin organs, paraganglia, and within the inferior mesenteric ganglion as well. They displayed the morphological character usually associated with their adrenal medullary catecholaminergic counterparts, including numerous dense-cored vesicles known to be the harbingers of catecholamines and various peptides. Nerve endings were not noticed upon the mitotic cells. The phenomenon of dividing extraadrenal chromaffin cells augments existing data and perhaps suggests that these cells are more endocrine than neural in type and subservient to the adrenal medulla in its classic endocrine function.
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  • 44
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    Journal of Electron Microscopy Technique 9 (1988), S. 283-291 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Kidney ; Artifacts ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The aim of this presentation is to draw attention to the problems inherent in evaluating the ultrastructure of percutaneous renal biopsies and to discuss some of the special techniques which are useful in this area. It is important to realize that the ultrastructure as it appears in this kind of material does not necessarily reflect conditions in vivo. Comparison with suitable reference material may, however, permit reliable conclusions in terms of pathological diagnosis and pathogenesis. It is advocated that purely qualitative methods, which until now have predominated in ultrastructure work with renal biopsies, be replaced by morphometry and semiquantitative methods when it is possible and practical to do so in any research situation.
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  • 45
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    Journal of Electron Microscopy Technique 10 (1988), S. 247-263 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Corpus striatum ; Pallidum ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The synaptic organization of the globus pallidus is reviewed with respect to present knowledge about neurons, fibers, axon terminals, and their intrinsic synaptic relationships. Information derived from studies employing Nissl stains, Golgi impregnations, lesion degeneration techniques, immunohistochemistry, and anterograde axonal labeling in various species are presented along with ultrastructural data. Studies indicate that the globus pallidus contains a principal efferent neuron with smooth or spiny dendrites and simple or complex terminal dendritic arborizations. This cell type receives convergent inputs from intrinsic and extrinsic sources and uses γ-aminobutyric acid as a transmitter. A smaller and separate population of pallidal projection neurons contains acetylcholine. Two other less frequent neuronal types, of small and medium size, have also been recognized. Three to six types of axonal boutons forming synaptic contacts with pallidal neurons have been recognized in various studies. Among these, three types (types I, II, and III) are the most prevalent. Studies indicate that the most frequent category (type I) originates from neostriatal neurons via radial fiber projections and contains immunoreactive GABA and enkephalins. The synaptic architecture of the globus pallidus is dominated by a mosaic-like arrangement of long dendrites that are ensheathed by longitudinally oriented axons making synapses en passant. Triadic synapses involving dendrites that are pre- and postsynaptic are encountered infrequently. Because both striatopallidal and pallidothalamic connections are inhibitory, pallidal target neurons in the thalamus may be “disinhibited” when the neostriatum is activated.
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  • 46
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    Journal of Electron Microscopy Technique 8 (1988), S. 137-158 
    ISSN: 0741-0581
    Keywords: Kaposi's sarcoma ; Lymphoma ; Histology ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Malignancies frequently arise in patients infected with human immunodeficiency virus (HIV), including those patients classified as having the acquired immunodeficiency syndrome (AIDS). Currently, Kaposi's sarcoma and certain types of lymphoma are considered to develop as a result of HIV infection, and other cancers have also been reported in these patients. For the most part, ultrastructural study of HIV-associated malignancies has been limited to Kaposi's sarcoma; the ultrastructural features of the epidemic form of this disease are generally the same as those of the classical form. The occurrence of these cancers in HIV-infected individuals appears to be related to the immunodeficiency caused by this virus, but the basic etiologic mechanisms remain unknown. In general, only palliative treatments are presently available for HIV-associated malignancies.
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  • 47
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    Journal of Electron Microscopy Technique 10 (1988), S. 187-204 
    ISSN: 0741-0581
    Keywords: Sympathetic nervous system ; Parasympathetic nervous system ; Enteric nervous system ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ultrastructure of synapses in the autonomic nervous system is reviewed. The synaptic organization of the parasympathetic ganglia is relatively simple. Preganglionic axons form synapses either on the soma or on short perikaryal processes of the ganglionic neurons. The presynaptic terminals have a cholinergic morphology and contain mainly small clear vesicles with a few large dense cored vesicles. A few neuropeptides have been localized to the large dense cored vesicles of these terminals. The postganglionic parasympathetic axons ramify within their target tissues where they form close associations, but not true synaptic contacts. Sites of release of transmitter are recognized morphologically as varicosities along the length of the axon that contain clusters of small clear vesicles with a few large dense cored vesicles. The organization of the sympathetic nervous system is somewhat more complex. In addition to acetylcholine, enkephalin also exists in these terminals, probably in the large dense cored vesicles. There are at least three types of ganglion cell neurons in the paravertebral portion of the sympathetic nervous system: those that contain norepinephrine alone, those that contain norepinephrine along with neuropeptide Y, and those that contain acetylcholine and vasoactive intestinal polypeptide. The first type provides innervation to the parenchyma of the target tissues, while the second mainly innervates blood vessels. The third type innervates the sweat glands. In the prevertebral ganglia, a fourth type of neuron exists that contains norepinephrine and somatostatin. This neuron probably innervates the gut. Preganglionic terminals of the cholinergic type form synaptic connections mainly with the dendrites of the sympathetic ganglion neurons. In addition to the types of synapses described for the paravertebral ganglia, neurons in the prevertebral ganglia receive synaptic connections from dorsal root ganglia and from the enteric nervous system. The sympathetic ganglia also contain interneurons that receive preganglionic synapses and form efferent synapses with some of the principal ganglion cells. The interneurons have been shown to contain a variety of transmitters, including norepinephrine, epinephrine, dopamine, serotonin, and a number of neuropeptides. The postganglionic sympathetic axons have a similar morphology to the parasympathetic axons. They form networks in their targets, and the axons display varicosities with concentrations of both small and large vesicles. After appropriate fixation, these vesicles are seen to possess dense cores. The morphology of the enteric nervous system has the greatest complexity. There are sensory neurons, interneurons and motor neurons of various types. In addition, it receives extrinsic connections from both the sympathetic and parasympathetic nervous system. The synaptic ultrastructure is correspondingly complex, with a large number of different, morphologically disticnt types of synaptic terminals. The function and transmitter content of these terminals is beginning to be elucidated.
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  • 48
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    Journal of Electron Microscopy Technique 7 (1987), S. 91-95 
    ISSN: 0741-0581
    Keywords: Lead poison ; Ultrastructure ; X-ray microanalysis ; Intranuclear inclusion ; Concretion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Intranuclear inclusions have been found in epithelial cells of renal proximal convoluted tubules (RPCT) in lead poisoned animals. It has been suggested previously that the inclusions contained lead. The present report gives evidence to the contrary. Samples of renal cortex were taken from adult rats fed laboratory chow containing 2% lead acetate for 1 month. Under the electron microscope (EM), intranuclear inclusions were observed in epithelial cells of RPCT. They were round or ovoid, and not delineated by a membrane. They consisted of two parts: Peripheral and central. Lysosomes increased in number, and many myelin figures appeared in the cytoplasm. In the lumen of the tubules, high electron-dense concretions were found. By x-ray microranalysis, neither the central part nor the peripheral part of the intranuclear inclusions produced a lead peak in the spectrum. Lysosomes and the cytoplasmic matrix also showed no lead peak. However, when the electron beam was focused on the concretion in the lumen of the RPCT, the Lα peak of lead (10.551 keV) and Kα peak of calcium (3.691 keV) were evident. From these findings it is suggested that intranuclear inclusions in the cells of RPCT are not lead deposits. Rather, it is the concretions found in the lumen of tubule that contain lead.
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  • 49
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    Journal of Electron Microscopy Technique 7 (1987), S. 17-27 
    ISSN: 0741-0581
    Keywords: Exoplasmic cell surface ; Protoplasmic cell surface ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Using rotary replication with platinum and carbon to embedment-free sections of polyethylene glycol (PEG)-embedded tissue from which PEG had been removed, the membrane specializations on the outer (exoplasmic) and inner (protoplasmic) cell surface as well as the organization of the cytoskeleton was demonstrated. The high quality and excellent preservation were comparable to that obtained by the rapid-freezing, deep-etched replica method. The present results indicate that the PEG method is essentially acceptable as a reliable morphological technique. Because either sectioned or replica images from the same tissue cells can be observed simultaneously, the PEG method with rotary replication should provide valuable information on cell ultrastructure.
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  • 50
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    Journal of Electron Microscopy Technique 7 (1987), S. 119-125 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Cytochemistry ; Megakaryoblastic leukemia ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Transmission electron microscopy, scanning electron microscopy, and ultrastructural cytochemistry were utilized to study megakaryoblastic cells from four patients suffering from megakaryoblastic leukemia. The results show that megakaryoblastic leukemic cells have a unique ultrastructural appearance, surface architecture, and cytochemical activity. The cells are positive for platelet peroxidase cytochemical reaction, which is localized in the perinuclear space and endoplastic reticulum, but not in the Golgi apparatus and cytoplasmic granules. They have a rather smooth surface and display blebs or tuberculi which are different from those in other types of leukemic cells as seen under the scanning electron microscope. The megakaryoblastic leukemic cells also show a special appearance under the transmission electron microscope, such as a cytoplasm which contains numerous small mitochondria, mostly concentrated in one pole of the cell. These ultrastructural and cytochemical characteristics of the megakaryoblastic leukemic cells revealed by the combined techniques are very useful in the diagnosis of megakaryoblastic leukemia.
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  • 51
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    Journal of Electron Microscopy Technique 6 (1987), S. 143-153 
    ISSN: 0741-0581
    Keywords: Mauthner cell ; Mixed synapses ; Gap junctions ; Inhibitory synapses ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Large myelinated club ending and small-vesicle bouton synapses on the distal part of the lateral dendrite of the goldfish Mauthner cell were investigated with thin section, freeze-fracture, and immunocytochemical electron microscopic methods. Large myelinated club endings form mixed synapses, having both gap junctions and chemical synaptic junctions. The correlation of the number of gap junction particles (connexons) and the data from electrophysiological studies of single large myelinated club ending synapses suggest that only a small fraction of gap junction channels are open at any given time during electrical synaptic transmission. The chemical synaptic junctions at the large myelinated club ending synapse have large, round synaptic vesicles, indicating that they are excitatory. This result is in agreement with electrophysiological data demonstrating the excitatory nature of this chemical synapse. Freeze-fracture of these excitatory chemical synaptic junctions reveals the presence of the intramembrane particle aggregates in the postsynaptic E face.Small-vesicle boutons form chemical synaptic junctions with small, flat or oval synaptic vesicles. These structural data, in combination with previous electrophysiological studies, suggest that the small-vesicle bouton synapses are inhibitory. In support of this theory, the cytoplasmic side of the postsynaptic membrane of some of these synapses show positive immunocytochemical reaction to monoclonal antibodies against the rat glycine receptor. Freeze-fracture data reveal intramembrane particle aggregates in the postsynaptic P face of some small-vesicle bouton synapses which could possibly represent glycine receptor aggregates.
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  • 52
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    Journal of Electron Microscopy Technique 6 (1987), S. 207-217 
    ISSN: 0741-0581
    Keywords: 3-D reconstructions ; Computer ; Graphics ; Tastebud ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have prepared software for producing three-dimensional reconstructions from serial micrographs using an IBM PC or compatible. The software can be configured for a variety of graphics board and digitizing tablet combinations.Data is entered into the program by digitizing contours directly from micrographs. The program can handle up to 2,000 contours per data file, of up to 255 object types. Morphometric information such as line length, perimeter, and area are generated for each contour. The reconstruction program aligns the plane information from each section and displays the final reconstruction on a high resolution (640 × 400 pixels) color monitor. Object types can be differentiated by line width, line color, and fill color. Hidden line processing and conditional fill routines make it possible to produce reconstructions with either a solid or semi-transparent appearance. Reconstructions can be generated quite rapidly from any viewing angle in the X, Y, and Z axes. The program has proven valuable for the elucidation of the three-dimensional nature of biological structures.
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  • 53
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    Journal of Electron Microscopy Technique 4 (1986), S. 147-156 
    ISSN: 0741-0581
    Keywords: Identified neuromuscular junctions ; Freeze fracture ; Active zones ; Synaptic efficacy ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We developed a technique for freeze-fracturing single physiologically identified neuromuscular junctions. This technique permits direct comparison of quantal content with morphological variables such as active zone length per unit terminal length for the same cell. The technique was developed to elucidate the structural basis for variability in transmitter release at the neuromuscular junction. The procedures were as follows: (1) record quantal content by conventional intracellular recordings; (2) mark cells for identification by fluorescent dye injection; (3) fix and stain endplate cholinesterase; (4) glycerinate and remove single fibers from the muscle; (5) draw endplate morphology; (6) freeze-fracture single muscle fibers; (7) examine in a transmission electron microscope; and (8) photograph and measure nerve terminal membrane ultrastructure. We found that approximately 15% of freeze-fractured single muscle fibers exhibited nerve terminal active zones. To demonstrate the usefulness of this technique, physiological and morphological information from an identified junction is presented. Freeze-fracture of identified cells has several advantages over thin sections, which cannot accurately show such things as active zone length, spacing, or intramembrane particles. This technique also has applications to the study of active zone ultrastructure in situations where neurotransmitter release is known to differ from normal levels. In addition, direct correlations between membrane structure and function can be studied in other preparations by this method.
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  • 54
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    Journal of Electron Microscopy Technique 2 (1985), S. 11-28 
    ISSN: 0741-0581
    Keywords: Ultrastructure ; Semithick sections ; Three-dimensional ; Serial sections ; Stereomicroscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Many transmission electron microscopes are available which can be used to examine biological material in 0.25-0.50-μm-thick sections. When compared to the traditional thin section, these “semithick” sections possess a number of inherent advantages: They can be screened for content with the phase contrast light microscope, they facilitate many types of studies requiring an analysis of serial sections, and they are frequently the optimum thickness for stereomicroscopy. Structures such as microtubule-associated components, as well as structural relationships between cellular constituents, may also be clearly visible in semithick sections which are not visible, or go unnoticed, in thin sections. Together these advantages enable an investigator to obtain a more complete three-dimensional picture of a cell or cell component in a significantly (i.e., up to 90%) shorter period of time than would be required if thin sections were used. Semithick sections may, therefore, make a study feasible which is not approachable, or which is approachable only with great difficulty, by conventional thin sectioning techniques.
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    Journal of Electron Microscopy Technique 2 (1985), S. 305-351 
    ISSN: 0741-0581
    Keywords: Tumor ; Diagnosis ; Electron microscopy ; Ultrastructure ; Pathology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The ultrastructural diagnosis of tumors requires a careful analysis that should be done in an orderly fashion. It requires precise planning from the time of specimen collection to the selection of the area to be examined. Pictures must be taken systematically and every micrograph should allow to answer whether the number of cells photographed is adequate; whether mitoses are present, what is the pattern of the tumor; what is the appearance of the cell membrane; whether the cells are joined by junctional complexes; whether free surfaces possess microvilli or cilia; what organelles are present and how they are distributed; whether there are secretory granules, melanosomes, or other cytoplasmic elements. Nuclear and nucleolar size and shape have to be taken into consideration. The composition of the interstitial extracellular matrix is important in certain types of tumors. Although these questions are not the only ones to be addressed, their use in a logical fashion is helpful when it concerns the ultrastructural diagnosis.
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  • 56
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    Journal of Electron Microscopy Technique 1 (1984), S. 199-201 
    ISSN: 0741-0581
    Keywords: Critical point drying ; Electron microscopy ; Ultrastructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The principles and methods for constructing an improved chamber for dehydration and critical point drying of multiple biological samples are described. The specimen chamber design is based on vertical positioning of the electron microscope grids or coverslips and permits minimal perturbation of laminar solvent flow past the specimens. This condition is requisite for optimal exposure of samples to solvents, which is necessary for complete dehydration and drying. Fragile samples, including chromosomes, critical point dried in the multisample chamber demonstrate crisp, well-preserved, three-dimensional morphology.
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