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  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 419-426 
    ISSN: 1059-910X
    Keywords: Sublimation drying ; Peldri II ; Tert-butyl alcohol ; Cell morphology ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The increasing importance of in vitro models has presented new challenges in SEM processing techniques. The present study has evaluated the quality of preservation of cultured human lens epithelial cells processed by critical point, Peldri II, and tert-butyl alcohol drying. Specimens processed by critical point drying produced specimens with severe cracking of cell processes and microcracks across cell membrane surfaces. Peldri II and tert-butyl alcohol drying eliminated breakage of the filopodia and lamellipodia as well as eliminating the microcracks across the apical membrane surface. The morphology of lens epithelial cells grown on Cytodex 3 beads appeared rounded with convoluted membrane surfaces. These morphological features were present for cells processed by all three methods. Cytodex 3 beads were subsequently shown to shrink 52% in diameter during dehydration, which results in an 89% reduction in volume for the bead. Cells grown on Biosilon beads, which do not shrink, had a morphology similar to the cells grown on a flat substrate. These results indicate that Peldri II and tert-butyl alcohol drying offer an attractive alternative to critical point drying when preparing cultured cells for SEM. Interpretation of cultured cell morphology must consider shrinkage of the substrate material as a possible contributor to artifact. © 1995 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 101-101 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 67-81 
    ISSN: 1059-910X
    Keywords: Transport ; Proteins ; Ions ; Micropuncture ; Tight junctions ; Permeability ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: To understand the process of sperm maturation, an understanding of interactions between the spermatozoa with the luminal fluid microenvironment and with the epididymal epithelium is necessary. The composition of epididymal luminal fluid of several species is well documented but the manner by which the epididymis contributes to the formation of this specialized milieu is not so well understood. A major role played by the epididymis is to finely regulate the movement of molecules into and out of the lumen. This ensures that as spermatozoa progress along the duct they are exposed to a continually changing, but optimal environment necessary for their maturation and survival. This review focusses on our current understanding of the contributions of the epididymal epithelium to the formation of a specialized luminal fluid microenvironment. The role of the blood-epididymis barrier, the composition of the epididymal luminal fluid, the permeability properties of the epididymal epithelium, and recent studies on a number of luminal fluid proteins and expression of the genes which encode these proteins are discussed. © 1995 Wiley-Liss, Inc.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 82-100 
    ISSN: 1059-910X
    Keywords: Vasectomy ; Epididymis ; Vas deferens ; Hydrostatic pressure ; Antisperm antibodies ; Spermatic granulomas ; Inflammation ; Lysosmes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Common principles can be discerned in the response of the epididymis to vasectomy, despite species differences. Increases in the size and number of lysosomes are the most frequent changes in the epididymal epithelium. The presence or absence of additional alterations such as changes in the height of the epithelium may be related to variations in distensibility of the vas deferens and epididymis. Direct measurements by micropuncture of epididymal and seminiferous tubule hydrostatic pressure indicate that, contrary to dogma, increased pressure in the distal epididymis after vasectomy is not generally transmitted to the seminiferous tubules. The epididymal interstitium shows microscopic changes indicative of chronic inflammation, with infiltration of macrophages, lymphocytes, and plasma cells, and rats with these lesions have higher antisperm antibody levels than animals lacking epididymal changes. Macrophages and neutrophils may enter the duct through the epididymal epithelium, at sites of rupture of the duct, and in the efferent ductules. Cyst-like spermatic granulomas occur in virtually all species where the epididymis or vas deferens ruptures with escape of spermatozoa. The sites and timing of granuloma formation may depend on the mechanical properties of the tract in different species, and they are probably important in the immune response to vasectomy. Postvasectomy sera in Lewis rats recognize a consensus repertoire of dominant autoantigens that closely resembles the antigens bound by sera from rats immunized with isologous spermatozoa. There are multiple routes for disposal of the sperm that continue to be produced after vasectomy. © 1995 Wiley-Liss, Inc.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 123-137 
    ISSN: 1059-910X
    Keywords: Sr-Cu-O system ; High-Tc superconductors ; Infinite-layer structure ; Planar defects ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The crystal chemistry of the alkaline-earth cupric oxides stabilized at high pressure was studied by high-resolution electron microscopy. Two families of superconductors, (Ca1-ySry)1-xCuO2 and Srn+1Cun O2n+1+δ (n = 1, 2), and one series of semiconductors, Srn-1Cun+1 O2n, are reported. Their structural characteristics are systematically interpreted on the basis of the simple infinite-layer structure seen in SrCuO2. Microstructures and their relevance to high-Tc superconductivity are discussed, particularly in the alkaline-earth-deficient infinite-layer phase. © 1995 Wiley-Liss, Inc.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 138-154 
    ISSN: 1059-910X
    Keywords: Copper oxycarbonates ; Layered structures ; Tc superconductors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 102-122 
    ISSN: 1059-910X
    Keywords: High Tc ; Superconductors ; Deformation modulations ; Oxygen ordering ; Superstructures ; YBCO ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A short overview is given of the possibilities of electron microscopy in the determination of the local, atomic scale structure of high Tc superconducting materials. Examples include the detection of weak oxygen ordering, description and characterization of deformation modulations in layered superconductors, and analysis of very long period superstructures. The ordering principles for tetrahedral chains in Ga-, Co-, or Al-substituted YBCO are discussed and their complex defect structures are described. The incommensurate modulation in YBCO-based materials containing SO4-tetrahedra, centered on the Cu(1) sites of the CuO-chain plane is attributed to the ordering of b-oriented SO4-rich chains in the Cu(1)-S-O layer; the structure is described in terms of an SO4-concentration wave. As examples of the new mercury-based superconducting family we discuss Y0.6Ca0.4Ba2Hg1-xMxCu2O6+y, which crystallizes in the space group P4/mmm with a = 0.3870(1) nm, c = 1.2537(1) nm. This cuprate belongs to the 1212 series; susceptibility measurements show a Tc (onset) of 90K, with a diamagnetic volume fraction of 27% at 4.2K to be reached. A second example is related to the compound Tl2HgBa4Cu2O10+y, in which ordering between single Hg layers and double Tl layers is observed. © 1995 Wiley-Liss, Inc.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 181-192 
    ISSN: 1059-910X
    Keywords: Electron Ronchigrams ; Numerical reconstruction ; Lens aberrations ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In a dedicated STEM instrument equipped with a field emission gun, shadow images are easily obtained and have many uses. They are very sensitive to misalignment of the instrument and astigmatism, and therefore can be used for rapid and accurate alignment of the microscope. For crystalline materials, the shadow image contains both the bright-field and dark-field images. It is a summation of the transmitted and diffracted beams, and is basically a kind of Gabor's in-line hologram. Under small or medium defocus, shadow images of a thin, well-orientated crystalline specimen take the characteristic form of Ronchigrams, which offer a unique means to calibrate the microscope operation parameters, such as the spherical aberration coefficient Cs and defocus settings of the objective lens, with high accuracy. With the calibrated values of Cs and δ, a transfer function of the objective lens may be generated. In the stage of numerical reconstruction, by adapting this transfer function to the experimentally recorded hologram the lens aberration introduced in forming the hologram may be corrected and an improved resolution may be achieved for electron microscope images. © 1995 Wiley-Liss, Inc.
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  • 11
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 31 (1995), S. 128-140 
    ISSN: 1059-910X
    Keywords: Actin ; Phospholipids ; Tight junctions ; Pancreas ; Testis ; Immunocytochemistry ; Fracture-label ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The fracture-label technique was used in conjunction with a monoclonal antibody to actin and the phospholipase A2-colloidal gold (PLA2-CG) method to examine the spatial distribution of actin filaments in relation to the three-dimensional arrangement of tight junctional strands in rat testes and exocrine pancreatic acinar cells. The intimate association of actin filaments with tight junctional strands in the pancreas and testis was also illustrated by a doublelabeling experiment in which freeze-fractured pancreas or testis was labeled with monoclonal antibody-protein A-gold (30 nm gold size) followed by incubation with a PLA2-CG complex (11 nm gold size). Freeze-fracture-exposed tight junctional strands in both testicular and exocrine pancreatic cells labeled by PLA2-CG complex indicated the presence of phospholipids in these cylindrical membranous structures. Immunolabeling of freeze-fractured testes with a monoclonal antibody to actin revealed a narrow band of gold particles juxtaposed to the cytoplasmic aspect of the protoplasmic membrane halves decorated with parallel linear arrays of cylindrical tight junctional strands. Many of the gold particles representing actin antigenic sites were in direct contact with the cross-fractured tight junctional strands. Fracture-label preparations of exocrine pancreas labeled with the monoclonal anti-actin antibody also exhibited a similar labeling pattern at the apex of acinars cells where the tight junction complex is located. Double-labeling experiments revealed the simultaneous labeling of actin and phospholipids in the same fracture-label preparations. Digestion of testicular and pancreatic tissue samples in a free PLA2 solution prior to labeling with the monoclonal antibody or PLA2-CG complex removed not only the gold labeling previously seen over the tight junctional strands but also reduced drastically the immunolabeling for actin that was previously seen associated with the tight junction complex. Taken together, results of the present study showed that actin filaments are structural components of the tight junction strands and are connected to the cytoplasmic aspect of the latter structures. The interaction between this particular cytoskeletal element and the tight junction may be through the binding of a special domain of the actin filament to the phospholipids that partially make up the tight junctional complex. © 1995 Wiley-Liss, Inc.
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  • 12
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 31 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 13
    Electronic Resource
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 193-207 
    ISSN: 1059-910X
    Keywords: Superconductivity ; n-type superconductors ; Nonstoichiometry ; Superconducting oxides ; T, T′ ; T* structures ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: After reviewing microstructural studies on superconducting materials showing T, T′, and T* structural types, results are presented on the microstructure of some n-type superconductors and related materials prepared with accurate control of the oxygen stoichiometry. Electron microscopy is used to describe the ordering of interstitial oxygen defects in T-type La2NiO4+δ leading to the formation of the n = 2 term of a homologous series with the general formula La8nNi4nO16n+1. Structural transitions and superstructure formation in the Pr2-x-yCexSryCuO4-δ system are reported, where T, T′:;, and T* phases are isolated as a function of both Ce and Sr content. © 1995 Wiley-Liss, Inc.
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  • 14
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 30 (1995), S. 208-217 
    ISSN: 1059-910X
    Keywords: Superconductors ; Electron energy loss spectrometry ; Transmission electron microscope ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Electron energy loss spectrometry (EELS) with a cold field emission gun (cFEG) transmission electron microscope (TEM) is implemented to analyze the evolution of the electronic structure and dielectric function of oxide superconductors. The O-K core loss spectra of p-type doped oxide superconductors are analyzed in terms of holes formation on oxygen sites, while low loss spectra are analyzed for free carrier plasmas, other spectral excitations, and their crystallographic confinement.It is illustrated that the transmission EELS with a cFEG TEM very much complement soft X-ray absorption spectroscopy and optical spectroscopy, with the added advantages of high spatial resolution (∼1-100 nm), and is compatible with other analytical, diffraction, and imaging techniques, which are readily available in a cFEG TEM. © 1995 Wiley-Liss, Inc.
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  • 15
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    Microscopy Research and Technique 30 (1995), S. 246-251 
    ISSN: 1059-910X
    Keywords: Bodonidae ; Cell surface ; Chemical fixation ; Cryofixation ; Quick freeze ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Freeze-fracture technique was used to analyse the structure of conventionally fixed and quickly frozen Bodo sp., a free-living kinetoplastid. In the former method, chemically fixed and cryopreserved cells presented a corrugated membrane pattern in the flagella and cell body surfaces. In the latter, however, replicas from quickly frozen unfixed flagellates showed membranes with a smoother aspect, allowing the observation of intramembranous particles (IMPs) on the fracture faces, hardly detectable in previously fixed samples. The IMPs were randomly distributed throughout the cell surface, except in the sparsely seen short IMP rows. © 1995 Wiley-Liss, Inc.
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  • 16
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    Microscopy Research and Technique 31 (1995), S. 63-78 
    ISSN: 1059-910X
    Keywords: Chitinase ; β-1,3-glucanase ; β-fructosidase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: During the past few years, cyto- and immunocytochemical techniques have been developed and widely used for locating and identifying various molecules in plant cell compartments. The last decade has witnessed tremendous improvements in molecular cytology, thus allowing an accurate in situ detection of various components thought to play important biological functions in the plant metabolism. The use of immunocytochemistry to investigate resistance mechanisms of plants upon pathogen attack has provided key information on the defense strategy that plants elaborate during a host-pathogen interaction. Of the various proteins induced in response to infection, chitinases and β-1,3-glucanases have been the focus of particular attention due to their believed antimicrobial activity through the hydrolysis of the main fungal wall components, chitin and β-1,3-glucans. Attention has also been paid to β-fructosidase, the enzyme that hydrolyzes sucrose into glucose and fructoside. The marked accumulation of this enzyme upon pathogen infection has led to the consideration that infection may greatly infleunce the metabolic activity of colonized tissues by creating alterations of source-sink relationships. Another facet of the plant's defense strategy that has been the focus of considerable interest is related to the accumulation of structural compounds, such as hydroxyproline-rich glycoproteins and callose, to reinforce the wall architecture, thus decreasing vulnerability to microbial enzymes. A number of alternatives designed to improve plant protection towards pathogen invasion have been suggested. Among these, the production of transgenic plants expressing constitutively a foreign resistance gene and the pretreatment of plants with elicitors of defense reactions have been the subject of intensive studies at the molecular, biochemical, and cytological levels. Results of such studies clearly demonstrate the important contribution that cyto- and immunocytochemical approaches can make to our knowledge of how plants defend themselves and how plant disease resistance can be directly enhanced. These approaches will undoubtedly be active areas for future research in the development of biological control alternatives in which the mode of action of the product used is of key importance. © 1995 Wiley-Liss, Inc.
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  • 17
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    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 31 (1995), S. 79-92 
    ISSN: 1059-910X
    Keywords: Immunogold technique ; Protein A-gold ; Peroxisome ; Subcompartment ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Peroxisomes, since their discovery as microbodies, have been studied mostly independently by electron microscopists and biochemists. The fine structure has been studied by electron microscopy, and the compositional enzymes and proteins by protein biochemistry. Electron microscopic histochemistry has been used to try to clarify the relationship between the fine structure and its constituents. The immunogold technique, a combination of electron microscopy and protein biochemistry, for the first time resolved this problem due to the high sensitivity and resolution power of the staining and the high reliability of the technique. The present paper reviews the way in which the immunogold techniques, especially the protein A-gold technique, revealed the localization of various enzymes or proteins in peroxisomes or peroxisomal subcompartments, and discusses why this technique should be employed in peroxisome research. © 1995 Wiley-Liss, Inc.
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  • 18
    ISSN: 1059-910X
    Keywords: Adenovirus ; Autoradiography ; Biotinylated probe ; Cytochemistry ; Electron microscopy ; Immunocytochemistry ; In situ hybridization ; Replication ; Transcription ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A significant amount of new information on structure-function relationships in nuclei of adenovirus-infected cells has accumulated during the last decade as a result of the combined use of several new cytochemical techniques. Localization of viral DNA on ultrathin sections of infected cells has been investigated at the ultrastructural level by using specific DNA staining and immunocytochemistry with monoclonal anti-DNA antibodies. Both techniques, however, concomitantly visualize cellular and viral DNA. The specific stain for DNA reveals the configuration of the DNA molecules in the different nuclear substructures, whateer their synthetic activities. The immunodetection of DNA reveals that specific antibodies strongly bind to DNA of condensed host chromatin and to both encapsidated and nonencapsidated inactive viral genomes. However, the observation of an abnormally low level of labeling over the substructures in which synthetic activities of viral genomes are known to be intense demonstrates a serious limitation of this technique for the detection of active DNA. Postembedding in situ hybridization is the most useful method for identifying with certainty the structures containing defined nucleic acid sequences. By using a biotinylated viral DNA probe, in situ hybridization provides specific identification of structures containing either viral DNA or viral RNA molecules. In addition, with appropriate pretreatment of the sections, it is possible to reveal either all the viral DNA-that is, both double- and single-stranded DNA molecules (dsDNA, ssDNA)-or more specific species such as only ssDNA or only dsDNA molecules. The replicative and transcriptional activities of viral genomes are determined by high-resolution autoradiography. Autoradiography after a short pulse incorporation of appropriate radioactive precursors by infected cells reveals the sites of cellular and viral DNA replication or trancription. A short pulse followed by chase periods of different durations reveals the progressive migration of the cellular and viral synthesized products. The in situ distribution of the viral 72 kDa DNA-binding protein, a highly phosphorylated protein which protects the viral ssDNA, is revealed either by immunocytochemistry with specific antibodies or by the bismuth staining method which stains all highly phosphorylated proteins, including both cellular and viral proteins. The combined results of all these cytochemical procedures reveal the composition and functions of some of the structures induced by adenovirus infection. They demonstrate that viral genomes engaged in replication lead to the formation of replicative foci in which two compartments rapidly develop, one of which results from the aggregation of single strands of viral DNA and their accompanying 72 kDa protein. Conversely, ssDNA and 72 kDa protein are rare in the other compartment which is the main site of replication and transcription of viral genomes. The procedural aspects and the contributions of electron microscope cytochemistry to an understanding of the biology of Ad5 viruses can serve as a basic framework for the study of other biological systems. © 1995 Wiley-Liss, Inc.
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  • 19
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    Microscopy Research and Technique 31 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 20
    ISSN: 1059-910X
    Keywords: Gastric mucosa ; Gap junction ; Tight junction ; Gastric cancer ; Gastric adenoma ; Gastric ulcer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Our aim was to determine whether the development of gap junctions in the human gastric mucosa has any relation to gastric ulcer and gastric carcinoma. Freeze-fracture replicas were prepared from the endoscopic biopsy specimens of 20 patients with gastric ulcer and 7 healthy volunteers. Large fractured areas of lateral cell membranes of surface mucous cells were examined randomly under an electron microscope. Small gap junctions were observed between gastric surface mucous cells in all healthy volunteers. Gap junctions in the patients with gastric ulcer were significantly fewer than in the healthy volunteers. In addition, gap junctions in patients with recurrent ulcer were significantly fewer than in those with first-onset ulcer. There was no obvious relationship between age and the development of gap junctions in patients with gastric ulcer or in healthy volunteers. In the areas of intestinal metaplasia, gap junctions were occasionally seen between absorptive cells of the villi, but not in the lateral membranes of goblet cells. Fresh frozen sections for indirect immunofluorescence were prepared from the endoscopic biopsy specimens of 19 patients with gastric ulcer and 5 patients with gastric cancer. Monoclonal antibody against liver gap junction protein (anti-connexin 32, 6-3G11) was used for the indirect immunofluorescence. On the border of gastric ulcer, fluorescent spots in the surface mucous cells were significantly fewer than in the surface mucous cells of the body and antrum which were distant from the ulcer area in the same patients. In gastric cancer tissue specimens, fluorescent spots were not observed at all. On the other hand, fluorescent spots in the noncancerous tissue of the patients with gastric cancer were present along the intercellular junctions between gastric surface mucous cells. These findings suggest that loss of intercellular communication via gap junctions is associated with gastric ulcer formation and gastric cancer formation. © 1995 Wiley-Liss, Inc.
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  • 21
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    Microscopy Research and Technique 31 (1995), S. 106-117 
    ISSN: 1059-910X
    Keywords: Acinar cells ; Duct cells ; Differentiation ; Immunogold ; Amylase ; Proline-rich proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Chronic treatment of mice and rats with isoproterenol (IPR) causes marked hypertrophy and hyperplasia of the salivary glands, and alters the expression of several secretory proteins. We used quantitative postembedding immunogold labeling to study the cellular responses in the rat parotid gland during daily (up to 10 days) injections of IPR and during recovery (up to 14 days) after cessation of IPR treatment. Labeling densities of acinar cell secretory granules with antibodies to amylase and protein SMG-B1 (cross-reactive with the rat homologue of Parotid Secretory Protein, PSP) fell to 10% of control levels after 8-10 IPR injections, then increased during recovery, paralleling previous biochemical determinations of changes in protein and mRNA levels. With antibodies to proline-rich proteins (PRP), labeling densities initially fell, then subsequently showed considerable variability, but never exceeded control levels. These results contrast with biochemical determinations showing a marked induction of PRP synthesis, and may have both immunological and structural explanations.Occasional intercalated duct cells located close to the acini underwent differentiation toward an acinar-like phenotype as a result of IPR treatment. After 1-2 IPR injections, the secretory granules of these cells labeled with antibodies to amylase and PRP. Subsequently, the granules appeared electron-lucent and were increased in size and number. These observations support earlier work, suggesting that intercalated duct cells may differentiate into other gland cell types.
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  • 22
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    Microscopy Research and Technique 31 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 23
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    Microscopy Research and Technique 31 (1995), S. 337-337 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
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  • 24
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    Microscopy Research and Technique 31 (1995), S. 338-346 
    ISSN: 1059-910X
    Keywords: Gap junctions ; Intercellular communication ; Cell-to-cell channels ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Gap junctions were discovered more than three decades ago, and since this time, enormous strides have been made in understanding their structure and function. This article summarises the part played by microscopy, within the context of multidisciplinary research, in the historical development of our knowledge of the gap junction. © 1995 Wiley-Liss, Inc.
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  • 25
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    Microscopy Research and Technique 30 (1995), S. 282-292 
    ISSN: 1059-910X
    Keywords: Steroid hormones ; Peptide hormones ; BPH ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Literature on the effect of steroid hormones (androgens, estrogens, and other steroids), of peptide hormones (e.g., prolactin), and growth factors (e.g., EGF, FGF, TGF-β), on the effect of castration and of experimental hormone application on the prostate is reviewed. Androgens have inductive, repressive, and interactive effects. They counterbalance an agonistic effect on proliferation and an antagonistic effect on cell death; they may influence DNA synthesis and induce the synthesis of substances with mitogenic effects on the prostate. Estrogens exert direct and indirect effects on the prostate. They suppress the secretion of gonatropins, thus repressing testicular androgen secretion. They stimulate the fibromuscular stroma and induce squamous metaplasia of the epithelium. Estrogens may also be involved in the onset of benign prostatic hyperplasia. Prolactin is preferentially bound in the diseased human prostate. An abundance of information has been gained on EGF, FGF, TGF-β, and other growth factors. They may be involved in the development of prostatic hyperplasia. Castration leads to a striking reduction in prostatic size in a short period of time due to autophagic and heterophagic processes. In castrated individuals, the prostate is enriched in androgen-independent cells. Experimental hormone application involves the substitution of androgens as well as anti-androgens, long-term application of different hormones, and application of combinations of drugs. The results of several studies are described. Further directions in the field of prostate research should concentrate on the role of growth factors in prostate development and pathology and on the effect of certain lectins on prostate diseases. We think that the investigation of interactions between steroid hormones and growth factors in normal and pathological neovascularization of the prostate is important. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 293-304 
    ISSN: 1059-910X
    Keywords: Testicular hormones ; BPH ; Cancer ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Testicular hormones regulate the growth and development of the prostate. The presence of androgen receptors in prostatic tissue and their importance in the normal development of the prostate has been established. Age-related changes in the hormonal milieu, and perhaps steroid hormone receptor profile, could set in motion pathological changes leading to the onset of benign prostatic hyperplasia (BPH) or prostate cancer which primarily affect older men. The accumulation of dihydrotestosterone with age, the reawakening of the inductive potential of the prostatic stroma, the altered rate of apoptosis with age, and the age-related changes in the ratio of testosterone: estrogen have all been implicated in the etiology of BPH. In addition to androgen receptors, several studies have documented the presence of estrogen and progesterone receptors in BPH and prostate cancer. So far, most studies have focussed on the correlation between the presence/absence of steroid hormone receptors and response to hormonal therapy. The molecular mechanisms by which these steroid hormone receptors regulate the onset or progression of BPH and prostate cancer are not yet clear. The chronological changes in the levels and distribution of steroid hormone receptors in normal prostatic tissue and the effect of such changes on the synthesis of growth factors, growth factor receptors, and oncogenes should be investigated. © 1995 Wiley-Liss, Inc.
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  • 27
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    Microscopy Research and Technique 32 (1995), S. 263-263 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 28
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    Microscopy Research and Technique 32 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 29
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    Microscopy Research and Technique 30 (1995), S. 319-332 
    ISSN: 1059-910X
    Keywords: Rat ; Prostate ; Epithelium ; Stroma ; Cytodifferentiation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Instructive influences of fetal mesenchyme were examined in heterotypic tissue recombinants consisting of urogenital sinus mesenchyme (UGM) from male and female rats and distal ductal tips from adult rat prostate. Tissues were grown under the renal capsule of male hosts for periods up to 28 days. Resultant growths exhibited typical prostate histology. Expression of lobe-specific proteins for the ventral (prostatic steroid binding protein [PSBP]) lateral (seminal vesicle secretion II [SVS II]), and dorsal prostate (secretory transglutaminase [TGase]) were examined by immunocytochemistry. Male or female UGM combined with terminal segments of the ventral or dorsal prostate and immunolabeled with antibodies to lobe-specific proteins demonstrated expression of all three secretory products. The pattern of staining was consistent with a compound inductive response from the UGM. Unique to this study was our ability to use a defined mesenchymal tissue (female ventral mesenchymal pad [VMP]). This tissue is specifically associated with ductal branching morphogenesis and cytodifferentiation of the ventral prostate. Distal ductal tips from the dorsal lobe of the adult male prostate when recombined with female VMP and grown in vivo exhibited transformation of secretory phenotype, and the epithelium expressed mRNAs for PSBP. Immunocytochemistry of serial sections did not demonstrate labeling for TGase in the new epithelial growth. Ultrastructural analysis of the heterotypic recombinants indicated that the epithelium had similar characteristics to those of normal ventral prostate. Early stages of the mesenchymal-epithelial interactions resulted in dedifferentiation of the adult epithelium to solid cords of stratified cells. These findings illustrate the potent instructive capacity of a defined fetal UGM to influence development and cytodifferentiation of adult prostate epithelium. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 342-350 
    ISSN: 1059-910X
    Keywords: Apoptosis ; BPH ; Growth regulatory factors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Homeostasis in the prostate is recognized to be maintained by a complex interplay between the opposing actions of cell proliferation and Cell death. Growth regulatory factors that promote or inhibit cell proliferation and promote cellular death have been identified in the prostate. The integration of these forces involves cellular cooperation between the prostatic stroma and epithelium. Hormone-regulated production of growth regulatory factors by one cell type may determine growth stimulation, inhibition, or cell death in a reciprocal cell partner. Imbalance between net cell proliferation and net cell death rates may result in abnormal growth leading to BPH. Additional study of the growth regulatory factors associated with distal vs. proximal epithelial cells and stroma and comparison of growth factor expression by the neonatal, postnatal growing, adult quiescent, and aging prostates will likely provide further insight into the regulation of prostate cell division and death. © 1995 Wiley-Liss, Inc.
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  • 31
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    Microscopy Research and Technique 31 (1995), S. 408-419 
    ISSN: 1059-910X
    Keywords: Neuronal mosaic ; Coupling ; Network ; Dopamine ; Nitric oxide ; Connexin ; Development ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The vertebrate retina is a highly laminated assemblage of specialized neuronal types, many of which are coupled by gap junctions. With one interesting exception, gap junctions are not directly responsible for the ‘vertical’ transmission of visual information from photoreceptors through bipolar and ganglion cells to the brain. Instead, they mediate ‘lateral’ connections, coupling neurons of a single type or subtype into an extended, regular array or mosaic in the plane of the retina. Such mosaics have been studied by several microscopic techniques, but new evidence for their coupled nature has recently been obtained by intracellular injection of biotinylated tracers, which can pass through gap junctional assemblies that do not pass Lucifer Yellow. This evidence adds momentum to an existing paradigm shift towards a population-based view of the retina, which can now be envisaged both as an array of semi-autonomous vertical processing modules, each extending right through the retina, and as a multi-layered stack of interacting planar mosaics, bearing some resemblance to a set of interleaved neural networks. Junctional conductance across mosaics of horizontal cells is known to be controlled dynamically with a circadian rhythm, and other dynamically-regulated conductance changes are also likely to make important contributions to signal processing. The retina is an excellent system in which to study such changes because many aspects of its structure and function are already well understood. In this review, we summarize the microscopic appearance, coupling properties and functions of gap junctions for each cell type of the neural retina, the regulatory properties that could be provided by selective expression of different connexin proteins, and the evidence for gap junctional coupling in retinal development. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 437-445 
    ISSN: 1059-910X
    Keywords: Intramembrane particle plaques ; Surface distribution ; Experimental manipulation ; Junction neogenesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Gap junctions provide the basis for the formation of elaborate networks of communication between cells in animal tissues. Electron microscopic examination of thin sections of plastic embedded gap junctions has provided valuable information on the anatomy and function of these remarkable structures. Freeze-fracture electron microscopy, however, has made available unique vistas of gap junction-bearing intramembrane surface-surface previously inaccessible to the researcher's eyes. Data on population density, distribution, size, geometry of intramembrane particle packing, and structural responses of gap junction components to experimental manipulation are simply and easily obtained with freeze fracture. Recent developments of sophisticated protocols of immunocytochemistry as applied to freeze-fracture replicas further serve to reinforce the notion that freeze-fracture is a powerful tool for study of gap junctions. Molecular techniques of gap junction gene transfection promise to add a truly unique dimension to investigations of the broad spectrum of functional roles of gap junctions. © 1995 Wiley-Liss, Inc.
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  • 33
    ISSN: 1059-910X
    Keywords: Gap junctions ; Connexin40 ; Connexin43 ; Conduction system ; Mammals ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Using immunohistochemical staining, the distribution of connexin40 (Cx40) and connexin43 (Cx43) was studied in rat, guinea pig, porcine, bovine and human hearts. These species display differences in the degree of morphological differentiation of the conduction system. This study was performed in the anticipation that comparison of the distributions of Cx40 and Cx43 in young and adult specimens may provide clues as to the physiological role of connexins in the heart. To a large extent, the distribution patterns of Cx40 and Cx43 are comparable between species. In neonates and adults, Cx43 was immunolocalized throughout the working myocardium, but in the conduction system Cx43 was detected only after birth. Cx40 was found to appear slightly earlier in development than Cx43 and to disappear when levels of Cx43 became more abundant. This time course was seen in working myocardium and in the ventricular conduction system. Together these data suggest that expression of Cx40 induces or facilitates expression of Cx43, while abundant expression of Cx43 in turn leads to suppression of Cx40 expression. The exceptions to this may represent blocks in this potential regulatory sequence. A second conclusion is that Cx40 and Cx43 containing gap junctions appear in the ventricular conduction system from distal to proximal and only after birth. This indicates that terminal differentiation of the conduction system occurs unexpectedly late in development. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 520-532 
    ISSN: 1059-910X
    Keywords: Perinuclear ; theca proteins ; Spermiogenesis ; Acrosome ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The perinuclear theca (PT) is a unique cytoskeletal element that encapsulates the mammalian sperm nucleus. It is divided into subacrosomal and postacrosomal regions. The objective of this study was to analyze and stage the intracellular distribution of several promiment bull PT proteins during spermatogenesis. For this purpose, polyclonal antibodies raised and affinitypurified against the 15.5-, 25-, 28-, 32-, 36-, and 60-kDa bull PT polypeptides were used as probes on sections of aldehyde-fixed testes. Immunoperoxidase staining revealed that the PT polypeptides first appeared early in spermiogenesis, concomitant with early steps of development of the acrosomic system. Immunogold labeling further showed that these polypeptides were peripherally associated with the entire acrosomal membrane, before and during the attachment of the acrosomic vesicle onto the spermatid's nucleus. Once the acrosome had capped the nucleus the labeling resided mainly in the subacrosomal region of the spermatid, between the inner acrosomal membrane and nuclear envelope. Later, during the elongation of the spermatid's nucleus, the labeling with all antibodies except the anti-15.5-kDa antibody extended caudally over the assembling postacrosomal sheath. This study suggests that the perinuclear theca proteins play an instrumental role in the attachment, spreading, and binding of the acrosome onto the nucleus of spermatids. © 1995 Wiley-Liss, Inc.
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  • 35
    ISSN: 1059-910X
    Keywords: Golgi apparatus ; Kinesin ; Sertoli cells ; Microtubules ; Cell polarity ; Tight junction ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Sertoli cells are polarized epithelial cells of the seminiferous epithelium which provide structural and physiological support for differentiating germ cells. They establish different basal and adluminal environments for the selective nurturing of pre- and post-meiotic germ cells within the seminiferous epithelium, segregated by the Sertoli-Sertoli cell tight junctional complex, the blood-testis barrier. Tight junction formation between epithelial cells in vitro is a critical polarizing event associated with changes in polarized targeting of membrane-specific proteins and reorganization of microtubules, centrioles, and the Golgi apparatus. To investigate whether tight junction formation is associated with organelle reorganization in Sertoli cells in vivo, we have characterized distribution patterns of Sertoli cell microtubules, the mechanoenzymes kinesin and cytoplasmic dynein, and the Golgi apparatus during tight junction formation in developing rat testis. Immunocytochemistry on samples taken at 5, 10, 15, 20, and 25 days of age was used to examine the distribution of these proteins during the extensive cellular reorganization that culminates in the formation of the blood-testis barrier at 19 days of age. Our data show that the distribution patterns reflect the extensive intercellular repositioning of tubule cells in developing seminiferous tubules, but that changes in intracellular organization are not temporally associated with formation of the blood-testis barrier. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 408-418 
    ISSN: 1059-910X
    Keywords: Mitosis ; Chromosomes ; Lung cells ; HeLa S3 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: There is general agreement that at the time of mitosis chromosomes occupy precise positions and that these positions likely affect subsequent nuclear function in interphase. However, before such ideas can be investigated in human cells, it is necessary to determine first the precise position of each chromosome with regard to its neighbors. It has occurred to us that stereo images, produced by scanning electron microscopy, of isolated metaphase plates could form the basis whereby these positions could be ascertained. In this paper we describe a computer graphic technique that permits us to keep track of individual chromosomes in a metaphase plate and to compare chromosome positions in different metaphas plates. Moreover, the computer graphics provide permanent, easily manipulated, rapid recall of stored chromosome profiles. These advantages are demonstrated by a comparison of the relative position of group A - specific and groups D - and G - specific chromosomes to the full complement of chromosomes in metaphase plates isolated from a nearly triploid human-derived cell (HeLa S3) to a hypo-diploid human fetal lung cell. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 467-468 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Topics: Natural Sciences in General
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    Microscopy Research and Technique 31 (1995) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 39
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    Microscopy Research and Technique 31 (1995), S. 446-451 
    ISSN: 1059-910X
    Keywords: Freeze-fracture ; Confocal microscopy ; Gap junction channels ; Mathematical analytical model ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In the heart, gap junctions electrically couple myocytes together. Electron- and light-microscope-based analyses have revealed that cardiac gap junctions show a variety of organizational patterns. At the level of gap-junctional channel aggregates, freeze fracture has demonstrated diverse channel packing arrangements in the membranes of different myocardial tissues. Ultrastructural and immunohistochemical studies have shown variation and specialization in the 3-dimensional spatial distribution of gap junctional contacts between different types of myocardial cells. Here, we estimate the access resistance of various configurations of gap junctions using physical principles and explore how certain of these specializations in gap-junctional organization may influence access resistance, a potentially important determinant of electrical conductance between coupled myocardial cells. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 452-466 
    ISSN: 1059-910X
    Keywords: Connexins ; Intercellular communication ; Crystallization ; Electron microscopy ; Image analysis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Cardiac gap junctions play an important functional role in the myocardium by electrically coupling adjacent cells, thereby providing a low resistance pathway for cell-to-cell propagation of the action potential. Two-dimensional crystallization of biochemically isolated rat ventricular gap junctions has been accomplished by an in situ method in which membrane suspensions are sequentially dialyzed against low concentrations of deoxycholate and dodecyl-β-D-maltoside. Lipids are partially extracted without solubilizing the protein, and the increased protein concentration facilitates two-dimensional crystallization in the native membrane environment. The two-dimensional crystals have a nominal resolution of 16 Å and display plane group symmetry p6 with a = b = 85 Å and γ = 120°. Projection density maps show that the connexons in cardiac gap junctions are formed by a hexameric cluste rof α1 connexin subunits. Protease cleavage of α1 connexin from 43 to 30 kDa releases ∼13kDa from the carboxy-tail, and the projection density maps are not significantly altered. Uranyl acetate stain penetrates the ion channel, whereas phospho-tungstic acid is preferentially deposited over the lipid regions. This differential staining can be used to selectively probe the central channel of the connexon and the interface between the connexon and the lipid. The hexameric design of α1 connexons appears to be a recurring quaternary motif for the multigene family of gap junction proteins. © 1995 Wiley-Liss, Inc.
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  • 41
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    Microscopy Research and Technique 31 (1995), S. 469-469 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 42
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    Microscopy Research and Technique 32 (1995), S. 437-448 
    ISSN: 1059-910X
    Keywords: Sexual differentiation ; Organ culture ; Sertoli cell ; Leydig cell ; Mesonephros ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The influence of mesonephric tissues and the extracellular matrix on mouse gonadal differentiation was examined in vitro. Gonadal ridges, with or without the adjacent mesonephric region, were removed from mouse embryos on day 12 post coitum (p.c.), and cultured in the presence or absence of reconstituted basement membrane (matrigel) for 5 days. Culturing control undifferentiated testes with mesonephric tissues induced normal testicular differentiation. When testes without mesonephric tissues were cultured in the absence of matrigel, testicular cord formation was not observed in the explants. Sertoli cells were irregularly arranged in the testicular parenchyma, and no continuous basal lamina was formed around the Sertoli cells. However, when testes without mesonephric tissues were embedded in matrigel and cultured for 5 days, the Sertoli cells were organized into testicular cord-like structures. The Sertoli cells positioned at the base of the cord-like structures were closely connected to the matrigel at their basal surface, and showed a polarized distribution of vimentin filaments in their basal cytoplasm. Leydig cells, on the other hand, were differentiated in all testicular explants. In all ovarian explants, germ cells normally entered meiotic prophase. Therefore, these findings indicate that the extracellular matrix permits testicular differentiation in the absence of the mesonephros, and that removal of mesonephric tissues leads to developmental failure of cord formation because the components of the extracellular matrix around pre-Sertoli cells are incomplete. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 457-458 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A miniature vise built into a 5 mm diameter copper capsule is described that holds small pieces of prefrozen, hydrated specimens at low temperatures within the lens of the Hitachi S900 high-resolution scanning electron microscope.
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    Microscopy Research and Technique 32 (1995), S. 498-503 
    ISSN: 1059-910X
    Keywords: Colocalization ; In situ hybridization ; Immunocytochemistry ; Testis ; Digoxigenin ; Transition proiein-1 ; Sulfated glycoprotein-1 ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Using testes fixed by perfusion with Bouin's fluid and embedded in paraffin wax, this study has established methods for combining in situ hybridization and immunocytochemistry on the same section to colocalize mRNA and protein for transition protein-1 (TP-1) and sulfated glycoprotein-1 (SGP-1), respectively. It was found that SGP-1 could be detected in tissue sections subsequent to the detection of TP-1 mRNA in situ. The finding that (1) the tissue pretreatments required to permeabilize the section and to allow access to the probe, and (2) the hybridization conditions themselves, had no adverse effect on the detection of antigen, eases the performance of this technique. On this basis, important information could be obtained on the transcriptional and translational activity of spermatogenic cells, if related probes and antibodies are utilized. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 459-497 
    ISSN: 1059-910X
    Keywords: Anamniotes ; Spermatocysts ; Amniotes ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Spermatogenesis appears to be a fairly conserved process throughout the vertebrate series. Thus, spermatogonia develop into spermatocytes that undergo meiosis to produce spermatids which enter spermiogenesis where they undergo a morphological transformation into spermatozoa. There is, however, variation amongst the vertebrates in how germ cell development and maturation is accomplished. This difference can be broadly divided into two distinct patterns, one present in anamniotes (fish, amphibia) and the other in amniotes (reptiles, birds, mammals). For anamniotes, spermatogenesis occurs in spermatocysts (cysts) which for most species develop within seminiferous lobules. Cysts are produced when a Sertoli cell becomes associated with a primary spermatogonium. Mitotic divisions of the primary spermatogonium produce a cohort of secondary spermatogonia that are enclosed by the Sertoli cell which forms the wall of the cyst. With spermatogenic progression a clone of isogeneic spermatozoa is produced which are released, by rupture of the cyst, into the lumen of the seminiferous lobule. Following spermiation, the Sertoli cell degenerates. For anamniotes, therefore, there is no permanent germinal epithelium since spermatocysts have to be replaced during successive breeding seasons. By contrast, spermatogenesis in amniotes does not occur in cysts but in seminiferous tubules that possess a permanent population of Sertoli cells and spermatogonia which act as a germ cell reservoir for succeeding bouts of spermatogenic activity. There is, in general, a greater variation in the organization of the testis and pattern of spermatogenesis in the anamniotes compared to amniotes. This is primarily due to the fact there is more reproductive diversity in anamniotes ranging from a relatively unspecialized condition where gametes are simply released into the aqueous environment to highly specialized strategies involving internal fertilization. These differences are obviously reflected in the mode of spermatogenesis and this is particularly true of the stage of spermiogenesis where the morphology of the species-specific spermatozoon is determined. Moreover, unlike amniotes, many anamniotes display a spermatogenic wave manifest, depending upon the species, either at the level of the cyst or seminiferous lobule.This variation in the organization of the testis makes certain anamniotes perfect models for investigating germ cell development and maturation. For instance, the presence of a spermatogenic wave provides an opportunity to manually isolate discrete germ cell stages for analysis of specific Sertoli/germ cell interactions. Furthermore, for many anamniotes, germ cells mature in association with a morphologically poorly developed Sertoli cell. This seeming independence of Sertoli cell regulation allows the in vitro culture of isolated germ cells of some species of anamniotes through several developmental stages. Thus, due either to the anatomical organization of the testis, or structural simplicity of the germinal units, nonmammalian vertebrates can provide excellent experimental animal models for investigating many basic problems of male reproduction. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 104-111 
    ISSN: 1059-910X
    Keywords: Microvascularization ; Stannius corpuscles ; Corrosion casting ; Scanning electron ; microscopy ; Blennius ; Gasterosteus ; Zosterisessor ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Blood supply and microvascular patterns of Stannius corpuscles were studied by scanning electron microscopy of vascular corrosion casts in the teleost fishes Blennius pavo, Zosterisessor ophiocephalus, and Gasterosteus aculeatus. Microvascular casts demonstrated that Stannius corpuscles - depending on their location - have an arterial supply derived either directly from the dorsal aorta, from the trunk of the first ventral segmental artery of the tail, or from a renal artery. Supplying arteries form a capsular capillary bed and a parenchymal capillary bed; both are composed of fine, freely anastomosing vessels with a homogeneous isotropic distribution. Central venules arise deep in the corpuscles. In the capsule, they form a single vein which drains into a segmental vein or directly into the caudal vein. Stanniocalcin, the hormone of the Stannius corpuscle, enters the renal circulation and reaches its main target organs, the gills, via posterior cardinal veins - heart - ventral aorta. Occasionally, some capsular venules empty into the trunk kidney peritubular venules. Capillaries are fenestrated and are embraced by pericytes with long, slender processes. The perivascular space contains collagen fibrils. Nerve fibers are found close to endothelial cells and pericytes. Vascular patterns of Stannius corpuscles are compared with those of the rat parathyroid glands and are discussed in respect to physiological implications. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 469-479 
    ISSN: 1059-910X
    Keywords: mpc ; DMD ; mdx ; Dystrophin ; Transgenic ; Gene transfer ; Retrovirus ; Adenovirus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Myoblast transfer therapy and gene therapy have both been proposed as potential treatments for inherited myopathies, such as Duchenne muscular dystrophy (DMD). The success of myoblast implantation in mouse models, where problems such as immune rejection are easily overcome, have led to similar experiments being attempted on Duchenne patients with limited, if any, success. Gene therapy, either by viral vectors or direct injection of the plasmid, has also had some success in animal models. Although both techniques, either separately or in combination, show some promise for the treatment of DMD, there are still many issues to be investigated in animal models, including the following: What is the best source of muscle precursor cells (mpc), and how may sufficient cells be obtained? What is the best vehicle for gene therapy? How far from the injection site can an implanted cell or gene have an effect? How can immune rejection of the injected cells or introduced protein be overcome? Does the introduced dystrophin lead to improved muscle function? Can cardiac muscle can be successfully treated by gene therapy? Can skeletal muscle which has undergone a great deal of damage be improved by either cell or gene therapy? © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 491-495 
    ISSN: 1059-910X
    Keywords: Heart ; “Section directed” cryosectioning ; Scanning electron microscopy ; Immunohistochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A new method to study the developing heart was developed. Using this “section directed” cryosectioning method, appropriate fixed embryos can be trimmed optimally to obtain sectional planes that, if necessary, can be matched with histologically treated sections. As a result, the morphological information obtained with the scanning electron microscope can be compared in detail with the information on the molecular phenotypes of the subpopulations of cells as deducted from staining patterns of the sections. This method allows combination of the specific advantages of sophisticated histological techniques, such as immunohistochemistry and in situ hybridisation, with those of the scanning electron microscope. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 480-490 
    ISSN: 1059-910X
    Keywords: Multiparametric image analysis ; Heart ; Myocardium ; Fetus ; Neonatal ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The study of the topological organisation of myocardial cells is a basic requirement for the understanding of the mechanical design of the normal and pathological heart. We developed a technique based on multiparametric image analysis of transmitted polarized light to generate maps of the azimuth and the elevation angles of the myocardial cells. The properties of birefringence of the myocardium embedded in methylmetacrylate were measured in papillary muscles with monitored 3D orientation. This birefringence is positive uniaxial with a 0° extinction angle when the axis of the fiber is parallel to the axis of the polarizer or the analyzer. Thick sections were studied between crossed polars, and four images of each section were digitized for an angle of the polarizer with the section varying from 0-67.5° in steps of 22.5°. The amounts of transmitted light for each setup of the polarizer were combined in order to extract the values of the azimuth angle (modulo 90°) and the elevation angle of the myocardial cells, according to the Johannsen equation. The respective maps of these angles were calculated and then assessed with confocal scanning laser microscopy. This method provides an efficient and accurate tool for the study of the histological architecture of the fetal and neonatal heart. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 496-512 
    ISSN: 1059-910X
    Keywords: 3D reconstruction ; Computer-aided reconstruction ; Database ; Serial sectioning ; Contour model ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper describes a structured approach for a standard setup of a computer program for 3D reconstruction from serial sections. Three-dimensional reconstruction as a technique increases in importance as, along with modern immunohistochemical techniques, it is a tool in the understanding of three-dimensional development patterns. In order to apply 3D reconstruction technique in a standard laboratory setup, an attempt was made to streamline the input and the manipulation of the data such that results are obtained easily. One will find a combination of two approaches in this paper: the first is a strict ordering of the complex data, and the second is an ordering of the processes that one wishes to apply on the data (together, these two approaches constitute an information analysis); because it was observed that developmental biologists tend to work from simple lines to describe their observations, the contour model was chosen as the vehicle to build a reconstruction model from. Consequently, the data is ordered in a database that has to be manipulated to get the data out in the desired format. The most important output format is a display of the reconstructed contour stack on a graphical computer screen. Together with the other data manipulation processes, such as the input, the inspection, the revision (correction), and the reconstruction, all processes are described using the reconstruction of an 11 embryonic days (ED) rat embryo as an example. Finally, the merits of the program are illustrated with an example from the development of the human embryonic heart. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 30 (1995), S. 513-520 
    ISSN: 1059-910X
    Keywords: Polyacrylamide embedding ; Confocal microscopy ; Specimen preparation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The preparation of optically clear, thick sections of fragile embryonic tissues greatly aids the power of confocal scanning laser microscopy in imaging three-dimensional structures. We report here conditions for embedding, sectioning, and staining embryos in polyacrylamide gels for a variety of confocal imaging techniques. Infiltration of tissues in standard mixtures of 10-15% acrylamide monomer yields, upon polymerization, blocks that cut easily by vibratome between 50 and 1,000 μm. These conditions worked well for tissues previously stained or for staining gel sections with low molecular weight water-soluble fluorochromes (MW 〈 5 kD [e.g., propidium iodide, phalloidin]). For immunostaining of tissue after embedding and sectioning, the acrylamide concentration was reduced to 2-3% acrylamide to allow access of immunoglobulins to antigenic sites; such gels were supplemented with 1% agarose to facilitate sectioning and handling. Either method yielded abundant, optically clear, and easily handled sections for mounting and examination in water-miscible media. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 521-530 
    ISSN: 1059-910X
    Keywords: Confocal light microscopy ; SEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Confocal light microscopy has found its place among the standard analytical tools in cell and molecular biology. When combined with techniques such as immunofluorescence or fluorescent in situ hybridization, the spatial distribution of individual biological components can be traced within cells and tissues and, under certain circumstances, even with living samples. In this article, advanced 3D visualization techniques have been applied to analyze the distribution of myofibrillar proteins in cultured adult rat cardiomyocytes. By combining confocal immunofluorescence microscopy with specially designed three-dimensional visualization, we have obtained images which are similar to those obtained with the scanning electron microscope. The subcellular distribution of proteins expressed after transfection of cDNA is monitored in the cultured heart cells. The expressed proteins are distinguished from their endogenous counterparts by the use of an epitope tagging technique. The described methods are suitable to specifically monitor the behavior of several closely related isoprotein mutants in cell or tissue preparations. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 1-3 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 31 (1995), S. 4-21 
    ISSN: 1059-910X
    Keywords: DNA ; RNA ; Cell nucleus ; Immunogold techniques ; Nuclear organization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In the present review, we report on recent results obtained by in situ transferaseimmunogold techniques as to the ultrastructural distribution of DNA and RNA within the cell nucleus. Special emphasis is placed on the various nucleolar components and the various enigmatic structures of the extranucleolar region: interchromatin granules, coiled bodies, and simple nuclear bodies. These data are discussed in the light of our current understanding of the functional organization of the cell nucleus. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 44-62 
    ISSN: 1059-910X
    Keywords: Mineralization ; Bone ; Cartilage ; Cementum ; Dentin ; Enamel ; Osteopontin ; Osteocalcin ; Bone sialoprotein ; Amelogenin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Immunocytochemistry is a powerful tool for investigating protein secretion, extracellular matrix assembly, and cell-matrix and matrix-matrix/mineral relationships. When applied to the tissues of bones (bone and calcified cartilage) and teeth (dentin, cementum, and enamel), where calcium phosphate-containing extracellular matrices are the predominant structural component related to their weight-bearing and masticatory roles, respectively, data from immunocytochemical studies have been prominent in advancing our understanding of mineralized tissue modeling and remodeling. The present review on the application of postembedding, colloidal-gold immunocytochemistry to mineralized tissues focuses on the advantages of this approach and relates them to conceptual, theoretical, and experimental data currently available discussing matrix-mineral interactions and extracellular matrix formation and turnover in these tissues. More specifically, data are summarized regarding the distribution and role of noncollagenous proteins in different mineralized tissues, particularly in the context of how they interface with mineral, and how this relationship might be affected by the various tissue-processing steps and immunocytochemical strategies commonly implemented to examine the distribution and function of tissue proteins. Furthermore, a technical discussion is presented that outlines several different possibilities for epitope exposure in mineralized tissues during preparation of thin sections for transmission electron microscopy. Cell biological concepts of protein secretion by cells of the mineralized tissues, and subsequent extracellular matrix assembly and organization, are illustrated by examples of high-resolution, colloidal-gold immunolabeling for osteopontin, bone sialoprotein, and osteocalcin in the collagen-based mineralized tissues and for enamel protein (amelogenin) in enamel. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 218-245 
    ISSN: 1059-910X
    Keywords: (Bi,Pb)2Sr2Can-1CunO4+2n+δ ; HR-TEM ; Crystal structure ; Microstructure ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: This paper summarizes results obtained by high-resolution transmission electron microscopy on the crystal structure and microstructure of the (Bi,Pb)2Sr2Can-1CunO4+2n+δ high-Tc superconducting oxides. The experimental basis for the work presented here are high-resolution structure images obtained at ultra-thin (3 nm) areas of carefully prepared transmission electron microscope (TEM) samples. The analysis was carried out on a 400 kV TEM equipped with a pole piece yielding 0.17 nm point-to-point resolution. From the images obtained the projected crystal potential of the cations can be extracted directly, as confirmed by detailed image simulation. Structural analysis of the oxygen sublattice remains an unsolved problem by high-resolution TEM (HRTEM), mainly because of the small scattering factors, and thus the contribution of the oxygen sublattice to the image contrast is small. The (Bi,Pb)2Sr2Can-1CunO4+2n+δ phases are modulated structures that can be understood as an average structure plus a superimposed displacement field. The crystal structure consists of BiO double layers and perovskite-type cuboids (containing Sr, Ca, Cu, and O), which are sandwiched between the BiO double layers. The displacement field can be directly analyzed by HRTEM, and the largest displacement amplitudes of 70 pm were determined for the Bi atoms in the n = 1 compound. The chemical composition of the n = 2 and n = 3 compounds was determined by EDX in the TEM for the cation sublattice. A significant (Ca + Sr) deficiency (approximately 10%) with respect to Cu was found. The (Sr + Ca)/Cu mole fraction ratio was 1.31 for the Bi-2212 phase and 1.14 for the Bi(Pb)-2223 phase. The oxygen content cannot be determined by EDX in the TEM with the accuracy necessary for a correlation with electrical and superconducting properties. The defect structure present in these materials, that is, intergrown lamellae with different crystal structures and equal or different chemical compositions, stacking faults, and grain boundaries, is summarized. The importance of grain boundaries for understanding and improving superconducting properties is emphasized. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 265-267 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 30 (1995), S. 252-257 
    ISSN: 1059-910X
    Keywords: Permeability tracer ; Endocytosis ; Transcytosis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Bovine aortic endothelial cells (BAEC) were exposed to glucosylated albumin-gold complexes (GgA), and the distribution of the tracers was compared after cryofixation and after glutaraldehyde fixation. Morphometric analysis revealed differences in the GgA distribution depending upon the method of fixation used. In BAEC monolayers cryofixed after 3 min of incubation with GgA, tracer was observed in predominately apically located vesicular elements. After 16 min of incubation, all vesicular elements were labelled, and multivesicular bodies were the prominent labelled structure. In contrast, chemically fixed monolayers exhibited a heterogeneous distribution of GgA within vesicular profiles after 3 min and 16 min of GgA incubation. The differences in tracer distribution depending upon the fixation method must be resolved before the mechanism of vesiclemediated endothelial cell transport function is defined and universally accepted. © 1995 Wiley-Liss, Inc.
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 31 (1995), S. 317-325 
    ISSN: 1059-910X
    Keywords: Cytochemistry ; Quantification ; EFTEM ; Teleost ; Tectum opticum ; Hair cell ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Four different methods for calcium precipitation are compared in the optic tectum and the inner ear of the cichlid fish, Oreochromis mossambicus. Several parameters are investigated concerning their influences on the reaction product. Three procedures (bichromate, fluoride, and oxalate-pyroantimonate) produce fine-grained deposits, often flocculent in the latter method. The fourth method (potassium-pyroantimonate) generates predominantly coarse-grained reaction product. The calcium content of the deposits is always proven with energy-filtering transmission electron microscopy (EFTEM). In both tissues fine-grained reaction product is found in endoplasmic reticulum and synaptic vesicles, and in addition in some mitochondria and at the cytoskeleton. The coarse-grained deposits of the potassium-pyroantimonate method have a more unspecific distribution. This is the only method which produces extracellular deposits in the inner ear, whereas in the optic tectum extracellular precipitates are always present except with the oxalate-pyroantimonate procedure. Two factors have an influence on the reaction product: the duration of fixation and the type of resin. The prolongation of the fixation time up to 24 hours leads to an increase of the reaction product, which also becomes coarse-grained. These observations are corroborated by quantification with image analysis. Furthermore the use of an epoxy resin compared to acrylic resins decreases the amount of reaction product produced. We show that the application of several methods is meaningful in order to understand the calcium properties of the investigated tissue, but it is necessary to optimize a certain method for a given tissue. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 326-333 
    ISSN: 1059-910X
    Keywords: Image analysis ; Automated focusing ; Extended focus ; Image focus ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Techniques to extract focus properties of microscopy images are many and varied. Many of them, however, rely on the summation of local statistical properties. Presented here is an examination of conventional focus determination algorithms, and a new method based on planar histograms of these local statistical properties. It is shown that this new method provides finer discernment of the focus properties of an image, and provides a means to extend the focus of an optical microscopy system. © 1995 Wiley-Liss, Inc.
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    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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    Microscopy Research and Technique 31 (1995), S. 118-127 
    ISSN: 1059-910X
    Keywords: In vivo ; Tubular epithelium ; Kidney ; Endocytosis ; Cationic albumin ; Immunocytochemistry ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The internalization and degradation of glomerular filtered serum proteins by the proximal tubular epithelium has been extensively studied by microperfusion methods. By using a cationic probe that easily traverses the glomerular wall into the urinary space, we have performed a morpho-cytochemical and quantitative study of the in vivo endocytotic activity of the proximal tubular epithelial cell. Bovine serum albumin (BSA) was tagged with dinitrophenol (DNP) and cationized to pI over 8. It was introduced into the circulation of normal mice for 5, 10, and 30 minutes and the distribution of the labeling was determined by protein A-gold immunocytochemistry, using specific antiDNP antibodies on tissue sections of routinely aldehyde-fixed, osmiumpostfixed, and Epon-embedded kidneys. Cationic BSA-DNP was detected at the endothelial and epithelial sides of the glomerular basement membrane, and over capillary and tubular basement membranes. In the proximal tubular epithelial cell, labeling was present over microvilli as well as over endosomal and lysosomal compartments, with labeling intensities varying from one compartment to the other. Morphometric evaluations of the labeling demonstrated a progressive incorporation of the probe from microvilli and endocytic compartments at 5 minutes to endocytic and lysosomal compartments at 10 and then 30 minutes. When considering labeling densities, no significant differences were found on microvilli and basolateral membranes between times of circulation; however, the labeling density over endosomal and lysosomal compartments was very intense at 10 minutes compared with 5 minutes, decreasing at 30 minutes. Results from this study validate the cationic albumin tagged with DNP as a tool in the study of the quantitative aspects of protein endocytosis at the ultrastructural level, in the kidney tubular epithelium. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 120-128 
    ISSN: 1059-910X
    Keywords: Parathyroid cell variants ; Fixation ; Membrane disintegration ; Secretory activity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Fixation with aldehydes is achieved either by immersion or perfusion. The parenchyma of parathyroid glands fixed by immersion consists of dark cells containing a lot of membranes of those organelles which are concerned with hormone secretion, light cells which are poor in these organelles, intermediate forms between the two, and multinuclear syncytial cells. They have been attributed to represent different functional stages of secretory activity, the dark cell being in an active form, the light cell in a resting form. Studies of the parathyroids of mice, rats, rabbits, cats, dogs, pigs, cattle, sheep, goats, and horses employing various fixation protocols clearly demonstrate that light cell variants and multinuclear syncytial cells are formed during improper immersion fixation as a result of membrane disintegration. Parathyroids fixed by perfusion or by immersion in an appropriate fixation medium comprise only one cell type which correspond to the dark chief cell. Parathyroid cells are polar cells bearing some of the rough endoplasmic reticulum in the basal pole, the rest of it, the Golgi complex, and secretory granules in the apical pole. The secretory product is released by exocytosis at the apicolateral domain of the plasma membrane into the intercellular space. Secretory activity can be altered experimentally, leading to drastic changes in the amount of cell membrane related to hormone synthesis, intracellular transport, exocytic release, and secretion coupled membrane retrieval. The sensitive reaction of parathyroid cells to both the mode of fixation and to fixation media demands careful evaluation of the fixation protocol. This and the polarity of parathyroid ceils have to be borne in mind for estimating secretory activity on the basis of morphological criteria. © 1995 Wiley-Liss, Inc.
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    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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    Microscopy Research and Technique 32 (1995), S. 267-285 
    ISSN: 1059-910X
    Keywords: MHC ; Class I ; Class II ; Brain ; Spinal cord ; CNS ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: MHC-restricted T cells are thought to contribute to clinical demyelination in MS and other circumstances. The step-by-step mechanisms involved and ways of controlling them are still being defined. Identification of the MHC+ cells in the CNS in situ has been controversial. This chapter reviews MHC expression in neural tissue, including normal, pathological, experimental, and developing tissue in situ and isolated cells in vitro. A basic pattern is defined, in which MHC expression is limited to nonneural cells and strongest class I and II expression are on different cell types. Variations from the basic pattern are reviewed. Ways of reconciling divergent findings are discussed, including the use of “mock tissue” to help choose between technical and biological bases for divergent findings, the potential contribution of internal antigen to the in situ staining patterns, and the possibility that class I upregulation is actively suppressed in situ. Functional implications of the observed patterns of MHC expression and ways of confirming the function of each MHC+ cell type in situ are described. It is suggested that modulating MHC expression in different cell types at different times or in different directions might be desirable. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 396-407 
    ISSN: 1059-910X
    Keywords: Gap junction ; Connexin ; Blood ; Haemopoiesis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: More than ten research groups have now reported the presence of gap junctions in blood-forming tissue or cultured cells. It is time to accept that these cell-coupling structures are present in this tissue. To find out what they are doing here we need to develop appropriate experimental techniques. This review covers the particular problems of investigating direct cell-cell communication by gap or other junction in undisturbed haemopoietic tissue. It then describes and assesses the published reports of haemopoietic gap junctions. Recently, in the author's laboratory, three means of increasing the number of gap junctions 50- to 100-fold in mouse marrow have been described, as well as techniques for doing so in culture. There is a complete report of this work here. At present it is quite unclear what function gap junctions serve in blood-formation, perhaps it is some consolation that 30 years after their ultramicroscopic discovery it is also true for all other unexcitable tissues. Possibly the ability to up-regulate their expression in haemopoietic tissue will help us find out what their role is in blood formation. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 295-301 
    ISSN: 1059-910X
    Keywords: Astrocyte ; Transplantation ; Remyelination ; Glia limitans ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Lesions in CNS white matter involving loss of glial cells with concurrent destruction of the glia limitans lead to widespread remyelination of CNS axons by Schwann cells. Previous studies have demonstrated that this situation can be changed by transplanting cultured CNS glial cells into lesions early on in the repair process. In this study we have transplanted cultured astrocytes into the sub-arachnoid space above such a lesion in order to (1) influence the normal repair process by transplant-assisted reconstruction of the glia limitans, and (2) explore the potential of a minimally invasive route for introducing cells to white matter lesions. In some cases, it proved possible to influence normal repair by transplanting cells via the sub-arachnoid route, although the results were inconsistent. However, the experiment permitted observations to be made on the migration of transplanted astrocytes across the surface of and within the spinal cord. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 302-311 
    ISSN: 1059-910X
    Keywords: JC virus ; In situ hybridization ; Progressive multifocal leukoencephalopathy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Progressive multifocal leukoencephalopathy is an important viral opportunistic infection of oligodendrocytes leading to direct demyelination. Virus is likely disseminated to the brain via the blood. However, the timing of that dissemination with relationship to clinical disease is unknown. Important clues about viral pathogenesis have been learned by applying molecular in situ techniques to diseased brain. The oligodendrocyte is the primary target for JC virus infection, and its death is the primary reason for demyelination. Bizarre astrocytes show limited viral DNA replication but are abortively infected. Although lymphoid organs can be infected by JC virus, there is no definitive evidence that lymphoid cells carry virus into the brain at the time of immunosuppression. JC virus may be reactivated from a latent state in both the brain and in non-central nervous system (CNS) organs at the time of immunosuppression, leading to clinical disease. Future sensitive in situ studies will likely resolve controversies about pathogenesis. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 204-214 
    ISSN: 1059-910X
    Keywords: Central and peripheral nervous systems ; Demyelination ; Remyelination ; Oligodendrocytes ; Schwann cells ; Globoid cell leukodystrophy ; Krabbe disease ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Twitcher mouse is an authentic murine model of human genetic demyelinating disease, globoid cell leukodystrophy (GLD), or Krabbe disease. Since its discovery at the Jackson Laboratory (Bar Harbor, ME) this model has been used extensively for the morphological, biochemical-enzymatic studies to clarify pathogenesis and also for therapeutic manipulation of genetic demyelinating disease in humans. As a result of these studies, now we know that (1) GLD is caused by a deficiency of lysosomal enzyme galactosylceramidase, and a toxic metabolite, psychosine, accumulates in the tissue, including the nervous system, damaging myelin forming cells and resulting in secondary demyelination; (2) morphological features of demyelination and associated cellular reactions in demyelination in this mutant are similar to those seen in autoimmune or toxic demyelination; and (3) with enzyme supplementation provided by bone marrow transplantation, remyelination occurs to some extent in demyelinated fibers in both central and peripheral nervous systems of twitcher mouse. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 30 (1995), S. 381-389 
    ISSN: 1059-910X
    Keywords: Electrophoresis ; Fibre type ; Isomyosin ; Myofibrillar actomyosin ATPase ; Myosin heavy chain ; Myosin light chain ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The present article attempts to combine existing information on the distribution of fast and slow myosin isoforms in histochemically distinct muscle fibres. Four myosin heavy chain (MHC) isoforms, MHCI, MHCIIa, MHCIIb, and MHCIId(x), have been identified in small mammals and have been assigned to the histochemically defined fibre types I, IIA, IIB, and IID(x), respectively. These fibres express only one MHC isoform and are called pure fibre types. Hybrid fibres expressing two MHC isoforms are regarded as transitory between respective pure fibre types. The existence of pure and hybrid fibres even in normal muscles under steady state conditions creates a spectrum of fibre types. The multiplicity of fibre types is even greater when myosin light chains are taken into account. A large number of isomyosins results from the combinatorial patterns of various myosin light and heavy chains isoforms, further increasing the diversity of muscle fibres. As shown by comparative studies, the distribution of different fibre types varies in a muscle-specific, as well as a species-specific manner. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 337-356 
    ISSN: 1059-910X
    Keywords: Olfactory signal transduction ; G proteins ; Adenylyl cyclase ; Rapid freezing ; Lowicryl ; Cilia ; Microvilli ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Freeze-fracturing and deep-etching are a well-suited set of methods to study membrane and cytoplasmic features. Various approaches are available. Possible variables include tissue preparation, fracturing only or fracturing followed by etching, modes and materials of replication, and various ways of combining freeze-fracturing and/or deep-etching with (immuno)cytochemistry. Freeze-substitution, in particular combined with embedding in methacrylate resins such as the Lowicryls, is becoming rather widely accepted for purposes of ultrastructural (immuno)cytochemistry. Most investigators active in this field agree that this combination yields superior results compared to (immuno)cytochemistry combined with more conventional means of thin section transmission electron microscopy. Yet relatively little information is available on the variations that can occur with different approaches of freeze-substitution immunocytochemistry. This review deals with some of the variations in freeze-fracturing, freeze-etching, and freeze-substitution as applied to olfactory epithelial structures and with the effectiveness of observations obtained by application of the above sets of methods in relating the special morphology of olfactory epithelial cellular structures with those obtained by other approaches. Indeed, the data obtained continue to provide an integral image in which that morphology can be related to the special biochemistry, cell and molecular biology, and electrophysiology of olfactory epithelial structures. © 1995 Wiley-Liss, Inc.
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  • 74
    ISSN: 1059-910X
    Keywords: Lactose repressor ; Mitochondrial ATPase ; F1 ; Freeze-etching method ; 45° unidirectional Pt-C replication of globular proteins ; 45° and vertical replication of gold ball standards on indirect carbon films ; Transmission electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The real size of platinum-carbon (Pt-C) replicated particles is not directly equivalent to either its metal-coated diameter or its shadow width. This paper describes two indirect methods, shadow widths and coated particle diameters, for determining a particle's actual size beneath a Pt-C replication film. Both produce equivalent measurements using the same standardized conditions: 2.3 nm Pt-C films deposited at a 45° angle on an ≈ -100°C surface in a 10 -6 torr vacuum. For the first method, gold balls nucleated in a partial pressure of helium and deposited on flat indirect carbon films (root mean square roughness of 0.8 nm) on 400 mesh grids were used as test particles for calibrating shadow widths as a function of particle size. The gold ball test specimens were replicated, and a distribution of Pt-C shadow widths orthogonal to the Pt-C deposition direction was measured and averaged for gold balls 1.5 ± 0.25 nm, 2.0 ± 0.25 nm, etc. The diameter of each gold ball was measured within the Pt-C film along with its shadow width because the Pt-C did not obscure or adhere well to the gold. The shadow width distributions for each gold size do not differ significantly from log normal. Two proteins, the lactose repressor and the mitochondrial ATPase, F1, were also used as replication test objects. Negative staining of both proteins was conducted to measure their average diameters. In the second method, a distribution of Pt-C-coated lac repressor diameters perpendicular to the shadow direction was measured. The Pt-C film thickness measured on the quartz crystal monitor was subtracted from the average metal-coated protein diameter to obtain the lac repressor's diameter. The Pt-C-coated particle diameter distributions also did not differ significantly from log normal. While doing this work it was discovered that outgassing the Pt-C electron gun greatly affected Pt-C film granularity: 19 sec produced a high contrast, granular Pt-C film, whereas 120 sec yielded a low contrast, less granular Pt-C film. Both gold balls and protein particles were subjected in separate experiments to either 19 or 120 sec of outgassing of the Pt-C gun prior to Pt-C replication. Outgassing had a profound effect on the average size of the Pt-C shadow widths on both gold and protein particles. The Pt-C gun outgassing procedure also determined the smallest replicated particle that could be resolved. The frequency of some smaller gold ball sizes detected after replication was reduced disproportionately with 19 sec vs. 120 sec outgassing. However, Pt-C gun outgassing did not affect the average measured diameter of the Pt-C-coated protein particles. The “geometric assumption” that each metal-coated particle creates a shadow width the same size as the metal-coated particle diameter was tested using a globular protein. Pt-C replication of protein particles at a 45° and 20° angle could not confirm the geometric assumption because an average shadow width was always significantly larger than its average Pt-C-coated particle diameter. A model for how the large shadow widths are formed is presented. Gold balls were also replicated at a 45° angle with current high resolution conditions at a substrate temperature of -185°C, and the results of these replicas were compared to the results reported here at ˜-100°C. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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  • 76
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    Microscopy Research and Technique 32 (1995), S. 363-363 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 32 (1995), S. 364-384 
    ISSN: 1059-910X
    Keywords: Spermatogenesis ; Spermatogenic cycle ; Seminiferous epithelium ; Germ cells ; Sertoli cells ; Testosterone ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Synchronous maturation of the germ cells in the seminiferous epithelium has long been recognized by microscopy, and is believed to be a consequence of a complex interaction between the germ cells and the Sertoli cells, largely driven by testosterone and its synergistic action with follicle-stimulating hormone. Overall coordination of the cycle of the seminiferous epithelium is reviewed with regard to the known and possible actions of testosterone upon the Sertoli cells and the germ cells. With gradual refinements of optical instrumentation and development of a wide range of histological, morphometric, biochemical, and molecular techniques, coupled with selective alterations of hormonal stimulation and the cellular composition of the testis, new approaches to the question of how sperm production is regulated are becoming available. Germ cell and Sertoli cell functions are intimately related to each other via local, intratesticular, or paracrine signals which are suppressed or triggered at certain defined steps in the spermatogenic process. The coordination of germ cell proliferation and maturation is discussed in terms of the contributions made by microscopical techniques. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 385-422 
    ISSN: 1059-910X
    Keywords: Spermatogenesis ; Germ cell degenerations ; Spermatogenic cycle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Spermatogenesis is a process of division and differentiation by which spermatozoa are produced in seminiferous tubules. A measure of efficiency of spermatogenesis is the estimated number of spermatozoa produced per day per gram of testicular parenchyma. This measure is not influenced by species differences in testicular size; however, it is influenced by species differences in the numerical density of germ cells and in the life spans of these cells. Seminiferous tubules are composed of somatic cells (myoid cells and Sertoli cells), and germ cells (spermatogonia, spermatocytes, and spermatids). Activity of these three germ cells divide spermatogenesis into spermatocytogenesis, meiosis, and spermiogenesis, respectively. Spermatocytogenesis involves mitotic cell division to increase the yield of spermatogenesis and to produce stem cells and primary spermatocytes. Meiosis involves duplication and exchange of genetic material and two cell divisions that reduce the chromosome number and yield four spermatids. Spermiogenesis is the differentiation of spherical spermatids into mature spermatids which are released at the luminal free surface as spermatozoa. The spermatogenic Cycle is superimposed on the three major divisions of spermatogenesis. Spermatogenesis and germ cell degeneration can be quantified from numbers of germ cells in various steps of development throughout spermatogenesis, and quantitative measures are related to number of spermatozoa in the ejaculate. Germ cell degeneration occurs throughout spermatogenesis; however, the greatest impact occurs during spermatocytogenesis and meiosis. Efficiency of spermatogenesis is related to the amount of germ cell degeneration, pubertal development, season of the year, and aging of humans and animals. Number of Sertoli cells and amount of smooth endoplasmic reticulum of Leydig cells (but not Leydig cell number) are related to efficiency of spermatogenesis. In humans, efficiency of spermatogenesis is reflected in number of spermatogenic stages per cross-section and number of missing generations within each stage; however, the arrangement of stages along the tubular length does not reflect differences in the efficiency of spermatogenesis. In short, spermatogenesis involves both mitotic and meiotic cell divisions and an unsurpassed example of cell differentiation in the production of the spermatozoon, and daily sperm production per g parenchyma is a measure of its efficiency. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 423-436 
    ISSN: 1059-910X
    Keywords: Stereology ; Number estimation ; Nuclear volume estimation ; Design-based methods ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The historical background to contemporary approaches to the estimation of cell/ nuclear number and volume in the testes is reviewed. The limitations of older geometric model-based approaches to the estimation of cell/nuclear number are discussed, and the need for absolute estimates of cell number rather than ratio estimates is examined. The physical and optical disector approaches to the direct estimation of numerical density and, hence, absolute cell number are presented together with data illustrating their operational efficiency in the testis. New approaches to the direct estimation of nuclear/cell volume, using the point-sampled intercept family of methods, are presented and illustrated, using the example of the Sertoli cell nucleus. The use of both classical transverse and the newer vertical section approaches is explored. Estimation of Sertoli cell/nuclear volume in the volume (point-sampled intercept procedure) and number (nucleator and rotator methods) distributions on both conventional transverse and vertical sections is discussed. The use of transverse sections of the testis is shown to produce a consistent bias in the estimation of Sertoli cell nuclear volume in 120-day-old animals, with all the estimators. Comparison of the Sertoli cell nuclear volume (measured on vertical sections) in the volume and number-weighted distribution suggests a coefficient of variation of volume in the number distribution of 0.4-0.5, suggesting either a random or stage-dependent variation in Sertoli cell nuclear size which requires further exploration. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 29-49 
    ISSN: 1059-910X
    Keywords: Mucus ; Mucous surfaces ; Glycocalyx ; Glycoproteins ; Estradiol ; Fallopian tube ; Oviduct ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The mucous surfaces of the oviducts of mammals, especially humans, are dependant on estradiol. The mucus glycoproteins and glycocalyceal glycoproteins have, however, barely been studied. Biochemical analyses have focussed on the relatively low molecular weight glycoproteins likely to be found in more serous-type granules of the ampulla and not on the very high MW glycoproteins typical of mucus and represented in the isthmus by morphological evidence of mucus secretion. Quantitatively, secretion from the ampulla is likely to predominate, because of its huge surface area compared with the isthmus. But functional closure of the isthmus under the influence of estradiol in the absence of progesterone means that it is the isthmus where luminal secretions accumulate - and where mucus glycoproteins will exert their most important effects on spermatozoa ascending the reproductive tract, and then on fertilized ova en route to the uterus. Further study of the extracellular, intraluminal, carbohydrate-rich environment of the oviductal isthmus, especially in humans, is likely to prove rewarding. Sampling of these secretions is now feasible using transvaginal, transuterine fallopian tube catheters that are in clinical use. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 13-28 
    ISSN: 1059-910X
    Keywords: Ciliated and secretory cells as related to menstrual cycle ; LM ; SEM ; TEM ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Oviductal epithelium of the baboon, Papio cynocephalus, was studied utilizing light, scanning, and transmission electron microscopy. Results of counts made of nonciliated, ciliated, and ciliogenic cells were analyzed statistically. The percentages of nonciliated cells of the fimbria and ampulla during the early proliferative and late secretory stages of the menstrual cycle were significantly greater than those during the mid-proliferative and late proliferative-early secretory stages, due to deciliation. This paper emphasizes previously unreported apical surface morphology as viewed by scanning and transmission electron microscopy. The microvillar pattern of the fimbrial secretory cells differs from that of the ampullar and isthmic cells in that the microvilli originate from thick apical protrusions and vary greatly in length and number as related to the cycle. A ridge demarcating the apical intercellular junction is composed of rows of microvilli during the early proliferative and late secretory stages. During the early proliferative and late secretory stages an increased degree of invagination of the basal and lateral plasma membranes occurs as the height and width of the cells decreases. The general numbers and distribution of the organelles of the various types of oviductal cells agree with that described for the ampulla and isthmus by Verhage et al. [(1990) Am. J. Anat., 187:81-90]; however, fimbrial epithelium was not included in that study. Other cyclic ultrastructural changes not examined previously include variation in the number of lipid droplets and their location, and in the number and relationships of glycogen particles to other structures. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995) 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 32 (1995), S. 75-76 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 32 (1995), S. 70-74 
    ISSN: 1059-910X
    Keywords: Electrolytic polishing ; Thin foils ; Specimen preparation ; TEM ; Dual-phase materials ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A two-stage jet polishing technique is described which, utilising the effects of the characteristic current-voltage behaviour of electropolishing solutions, can produce excellent TEM foils of relatively coarse two-phase materials. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 57-69 
    ISSN: 1059-910X
    Keywords: Oviduct ; Glycoprotein ; Antibody ; cDNA ; Human ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The major objective of this study was to examine the hormonal regulation of a human oviduct-specific glycoprotein (huOGP) throughout the menstrual cycle and in all regions of the human oviduct. Regulation of synthesis and secretion was examined at both the protein (Western immunoblots and immunocytochemistry) and mRNA (Northern and slot blots) levels and correlated with changes in the morphological features of the oviductal epithelial cells throughout the cycle. Immunoblot analysis of oviductal fluid and explant culture media from all regions of the oviduct demonstrated that huOGP is primarily found during the follicular stage of the cycle and is not present in serum, follicular fluid, or uterine endometrium. Moreover, two-dimensional (2-D) immunoblots showed that all major isoelectric variants of huOGP observed on 2-D fluorographs are immunologically related. Light microscopic immunocytochemistry localized huOGP to oviductal secretory cells in both ampulla and isthmic regions, with the most intense immunoperoxidase staining seen in midcycle samples. Using an indirect immunogold technique at the electron microscopic level, huOGP was specifically localized to secretory granules of the ampullary and isthmic nonciliated epithelial cells. The ultrastructural characteristics of these secretory cells during the mid to late follicular phase of the cycle suggested elevated protein synthetic activity. In addition, mRNA expression for huOGP was elevated in all regions of the oviduct in midcycle specimens. Collectively, these data indicate that huOGP is a major tissue-specific, stage-specific secretory product of the human oviduct during the periovulatory stage of the cycle and support the hypothesis that huOGP synthesis and secretion may be regulated by fluctuations in the levels of estrogen and progesterone. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 32 (1995), S. 77-78 
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    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
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    Microscopy Research and Technique 32 (1995), S. 79-90 
    ISSN: 1059-910X
    Keywords: Calcium ; Phosphate ; Urodela ; Anura ; Parathyroidectomy ; Parathyroid hormone ; Prolactin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Amphibians living partially or totally in a terrestrial environment are the first tetrapods to possess parathyroid glands. Purely aquatic amphibians and amphibian larvae lack these endocrine glands. The parathyroids develop at the time of metamorphosis. The parathyroid glands in caecilians consist of a single cell type, that of urodeles may be composed of basal (supporting) cells and suprabasal (chief) cells, and that of anurans of small and large chief cells. Parathyroid glands of caecilians and anurans lack connective tissue, blood vessels, and nerves. The parathyroid cells become activated in response to decreased blood calcium concentration and undergo changes indicating increased parathyroid hormone secretion. Increased blood calcium concentration suppresses secretory activity. Usually, parathyroidectomy elicits hypocalcemia in most amphibians. Such operations have no effect in lower urodeles. Parathyroid hormone administration provokes hypercalcemia in most amphibians. The parathyroids of caecilians have not been studied in detail. The urodeles and anurans exhibit seasonal changes in the parathyroid glands. These changes may be initiated by environmental stimuli such as light, temperature, or alterations in blood calcium levels caused by natural hibernation. © 1995 Wiley-Liss, Inc.
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  • 88
    ISSN: 1059-910X
    Keywords: Chelonia, Crocodilia ; Rhynchocephalia ; Squamata ; Parathyroid hormone ; Development ; Seasonal variation ; Parathyroidectomy ; Topography ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Adult reptiles possess one or two pairs of parathyroid glands that have been shown in many species to derive from the third and fourth pharyngeal pouches, respectively. Up to five pairs may develop during early embryonic life. Excess glands may involute during late embryogenesis. The location of the parathyroid glands differs in the various species. As a general rule, they lie just anterior to the heart, the anterior pair (parathyroid III) being associated with the carotid artery, the posterior pair (parathyroid IV) with the aortic arch. In snakes, however, the anterior pair (parathyroid III) is associated with the carotid artery near the angle of the jaw. As shown by light microscopy and, to a lesser extent, by electron microscopy, the parathyroid parenchyma comprises secretory cells which may form dark and light variants, occasional oxyphil cells, and stellate cells. They are arrangend in cords separated by connective tissue containing a capillary network. Parathyroid secretory cells often form follicles which might be the result of degeneration. Degeneration may occur as a form of involution during winter in species undergoing seasonal changes. The product of parathyroid cells, the parathyroid hormone, is responsible for the maintenance of blood calcium concentration. The sites of action - bones, kidneys, intestine, endolymphatics, and dermal skeleton - are not well understood or not investigated. In some turtles, parathyroid hormone is not the (main) factor for the regulation of calcium homeostasis. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 275-284 
    ISSN: 1059-910X
    Keywords: Ultramicrotomy ; Cross-sections ; Epilayers ; Semiconductors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ultramicrotomy has found novel application in materials research using transmission electron microscopy, for both analytical and high resolution work. Specimens 20-50 nm in thickness with lateral dimensions of up to 100 microns may routinely be prepared, with the additional advantage of precise area location for cross-sectional samples from real devices (e.g., VLSI structures). Much of this work is possible through the availability of diamond knives with various included angles and lengths at reasonable cost. This paper reports on various applications of ultramicrotomy, in particular, lattice imaging of surfaces and interface regions from epilayers of II-VI compound semiconductors and related materials. Ultramicrotomed cross-sections have enabled modern electron beam imaging, diffraction and analytical techniques to be brought to bear near surfaces and across interfaces of multilayer structures, yielding high spatial resolution information on crystallography, defect structure and composition. © 1995 Wiley-Liss, Inc.
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    Microscopy Research and Technique 31 (1995), S. 293-299 
    ISSN: 1059-910X
    Keywords: Sample preparation for ATEM ; PEELS imaging ; Zn-Fe intermetallics characterization ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The formability of galvanneal steel sheets used in the automotive industry is influenced by the presence and distribution of brittle and difficult to distinguish Zn-Fe intermetallics in the coating. Characterization of these intermetallics requires a high spatial resolution technique such as analytical transmission electron microscopy (ATEM). Sample preparation by ion milling is impossible due to iron redeposition, and traditional ultramicrotomy using water affects the coating chemistry. A technique based on dry ultramicrotomy has therefore been developed.To optimize the technique, different parameters (knife angle, cutting medium, thickness setting on the ultramicrotome, cutting speed) have been investigated for the preparation of galvanneal coatings and pure Al sections. Results show that dry cutting does not affect the coating chemistry but shortens the life of the knife. Knife quality (cleanliness, sharpness and absence of defects) is a major factor to obtain good dry sections. The best results for the more ductile pure Al are obtained with a 35° knife whilst for the harder galvanneal coating it is recommended to use a 55° knife. These results suggest that the sectioning mechanism for the harder material involves more a cleavage-fracture mechanism whilst a greater amount of shear is involved when sectioning relatively ductile Al. The optimum parameters for sectioning galvanneal coatings are established and results obtained by parallel electron energy loss spectrum imaging and energy dispersive X-ray spectrometry in the TEM are given. This study shows that with a good control of all the sectioning parameters it is possible to obtain good sections repeatedly and rapidly. © 1995 Wiley-Liss, Inc.
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  • 91
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    Microscopy Research and Technique 31 (1995), S. 285-292 
    ISSN: 1059-910X
    Keywords: Oxide film ; Barrier film ; Ultramicrotomy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A recent advance in metallurgical technology has been the application of rapid solidification techniques to Al alloy production. FVS0812 is the designation given to a microcrystalline Al-based alloy consisting of 8 wt% Fe, 1 wt% V and 2 wt% Si. It is a two-phase alloy, consisting of ca. 27 vol percent of approximately spherical Fe-V-Si-rich dispersoids in an essentially pure Al matrix. The high strength, low density properties of this advanced material, and other related alloys, have not yet been realized, however, due, in part, to the inability of the alloy to form a thick, adherent, abrasion-resistant outer surface oxide film, a feature readily achieved at conventional Al alloys by normal anodizing methods. The present research has involved an electro-chemical study of oxide film growth at the 812 alloy, with the specific goals being to seek an understanding of the origin of the oxide film growth problem and ultimately to propose alternative approaches to the formation of a thick, stable oxide film at this material. The techniques used in this research have included electrochemical methodologies such as cyclic voltammetry and electrochemical impedance spectroscopy. Crucial information has been obtained through transmission electron microscopy (TEM) of ultramicrotomed specimens. Experiments were carried out initially in neutral borate solutions to characterize the compact barrier oxide film formed in this environment and expected to be present beneath the porous oxide film formed in the normal sulfuric acid anodizing medium. In borate solutions, the electrochemical results implied oxide film thicknesses which were less than seen subsequently by TEM work, suggesting either that the barrier film at the 812 alloy can be penetrated by solution in very fine pores (not resolvable by conventional TEM) at its outer surface or that dispersoids trapped in the oxide film cause differential oxide film thicknesses to develop across the alloy surface. In sulfuric acid solutions, dissolution of Fe and V occurs from the 812 alloy during anodization. Both impedance and TEM studies reveal the absence of a barrier film at the 812 alloy surface. Also, the thick oxide overlayer has a tortuous and more open pore structure than formed at Al and the oxide film is also substantially thinner than it should be. It is suggested that the absence of a barrier oxide film indicates that the sulfuric acid anodizing medium is too aggressive for oxide film formation at the 812 alloy, resulting in excessive dissolution and poor oxide film qualities. © 1995 Wiley-Liss, Inc.
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  • 92
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    Microscopy Research and Technique 31 (1995), S. 308-310 
    ISSN: 1059-910X
    Keywords: Thin films ; TEM specimen preparation ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ultramicrotomy has been used to prepare TEM cross-sections of typical hard dielectric, semiconductor, and metal coatings, providing a critical capability in the study of structure-property relationships of thin films. Ultramicrotomy of thin film coatings requires meticulous attention to technique and handling. The sample to be microtomed must be very small, well bonded to the epoxy embedding medium, and precisely oriented. In this article we report the ability to microtome TEM cross-sections of diamond and cubic boron-nitride (cBN) coatings. © 1995 Wiley-Liss, Inc.
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  • 93
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    Microscopy Research and Technique 31 (1995), S. 300-307 
    ISSN: 1059-910X
    Keywords: TEM ; Ion beam ; Chemical jet ; Diamond knife ; SAED ; Alumina ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Industrial materials, such as alumina, often pose difficulties in their preparation for TEM examination. Composite and particulate materials are particularly difficult to prepare using conventional thinning techniques, i.e., ion beam and chemical jet thinning. Ultramicrotomy (UM) can be used to produce TEM specimens with a uniform thickness and an unaltered composition.Some crystalline materials, i.e., alumina hydrate, were difficult to section due to conflict between the cutting direction and cleavage planes. Sectioning was successful when these two directions were mutually parallel or perpendicular. At other orientations shattering occurred. Microcrystalline particulate materials, i.e., calcined alumina, were sectioned successfully in particles 〈 70 μm in diameter.The phases found in industrial alumina particles were gamma, delta, theta, and alpha alumina. Gamma alumina consisted of fine-grained, equiaxed crystallites. The selected area electron diffraction (SAED) patterns indicated poor crystallinity with a distinct hexagonal texture. Delta and theta alumina appeared as an undifferentiated intermediary microstructure of elongated grains. The SAED patterns indicated poor crystallinity, but without a distinct texture. Alpha alumina was found to be a coarse-grained crystalline phase with high diffraction contrast. SAED patterns consisted of fine, randomly oriented spots.Considerable variation was observed in the distribution of phases. In some specimens, discrete particles of gamma and alpha predominated. In others, particles were a mixture of phases repre-sentative of the bulk composition. To characterise these samples, TEM of numerous whole particles was required. Ultramicrotomy was the only preparation technique capable of producing such samples. © 1995 Wiley-Liss, Inc.
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  • 94
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    Microscopy Research and Technique 31 (1995), S. 193-214 
    ISSN: 1059-910X
    Keywords: Mouse stomach ; Cell migration ; Parietal cell ; Stem cell ; Zymogenic cell ; Surface mucous cell ; Mucous neck cell ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The secretions of the mammalian stomach are produced by cells present in invaginations of the epithelium, which in the mouse are straight tubules referred to as “zymogenic units.” These units comprise four regions, namely pit, isthmus, neck, and base, in which there are several cell lineages with different phenotypes and migratory pathways. In the isthmus, stem cells designated “undifferentiated granule-free cells” undergo division so as to maintain their own number and produce several differently oriented progenitors: (1) “Pre-pit cell precursors” are characterized by prosecretory Golgi vesicles with a uniform, fine particulate content. They give rise to “pre-pit cells” defined by the presence of few dense mucous granules. These cells migrate outward from the isthmus to the pit, where they become the dense granule-rich “pit cells” which populate the pit region and migrate to the gastric surface where they are lost. (2) “Pre-neck cell precursors” are identified by prosecretory Golgi vesicles containing an irregular dense center and a light rim. They give rise to “pre-neck cells” defined by a few mucous secretory granules with a clear-cut core. These cells migrate inward from the isthmus to the neck where they become “neck cells,” which contain many such granules. Even though neck cells are mature mucus-producers, they are not end cells. As they enter the base region, they become “prezymogenic cells” whose phenotype gradually changes from mucous to serous. These cells eventually lose the ability to produce mucus and thus become the typical zymogenic cells that populate the base region. (3) “Pre-parietal cells” are classified into three variants, which probably come from three different sources, that is, pre-pit cell precursors, pre-neck precursors, and the undifferentiated granule-free cells themselves. The preparietal cells mature into parietal cells which migrate either outward to the pit or inward to the neck and base. As a result, parietal cells are scattered in the four regions of the unit. (4) Precursors of “entero-endocrine” and “caveolated” cells give rise in the isthmus to these cells, which may also migrate outward or inward. © 1995 Wiley-Liss, Inc.
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  • 95
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    Microscopy Research and Technique 31 (1995), S. 93-94 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 96
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    Microscopy Research and Technique 31 (1995), S. 95-105 
    ISSN: 1059-910X
    Keywords: Striated muscle ; Smooth muscle ; Antibodies ; Localization ; Colloidal gold ; Proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The muscle cell cytoskeleton is defined for this review as any structure or protein primarily involved in linking or connecting protein filaments to each other or to anchoring sites. In striated muscle, the M line connects thick filaments at their centers to adjacent thick filaments. Titin forms elastic filaments that extend from the M line to the Z line and may contribute to the resting tension properties of striated muscle. Nebulin forms inextensible filaments in skeletal muscle that are closely associated with thin filaments and that may provide a length template for thin filaments. Z lines anchor thin filaments from adjacent sarcomeres via the actin-binding function of α-actinin. Other proteins located at the Z line include Cap Z, Z-nin, Z protein, and zeugmatin. Intermediate filaments connect myofibrils to each other at the level of the Z line and to the sarcolemma at the Z- and possibly the M-line levels. Immunolocalization has identified the adhesion plaque proteins spectrin, vinculin, dystrophin, ankyrin, and talin at subsarcolemmal sites where they may be involved with filament attachment. Smooth muscle cell cytoskeletons are believed to include membrane associated dense bodies (MADBs), intermediate filaments, cytoplasmic dense bodies (CDBs), and perhaps a subset of actin filaments. MADBs contain a menu of attachment plaque proteins and anchor both thin filaments and intermediate filaments to the sarcolemma. CDBs are intracellular analogs of striated muscle Z lines and anchor thin filaments and intermediate filaments. © 1995 Wiley-Liss, Inc.
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  • 97
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    Microscopy Research and Technique 31 (1995) 
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 98
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 99
    ISSN: 1059-910X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
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  • 100
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    Microscopy Research and Technique 31 (1995), S. 257-264 
    ISSN: 1059-910X
    Keywords: Cell count ; Cell division ; Computer-assisted image processing ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The development of computer-assisted image analysis has provided the technology to rapidly determine the population size of cultured cell monolayers in situ. We have adapted this technology to determine the population growth rate of cultured fibroblasts for use in a highreplicate format. Human lung fibroblasts were seeded into 1/2 A 96-well plates that had one-half the culture area of standard 96-well plates. The cells were cultured in medium supplemented with different concentrations of FBS and on days 0, 1, 2, 3, 5, and 7, and their nuclei were stained with propidium iodide. A microscopic field representing one-quarter of a well of fluorescent nuclear images was captured onto a Macintosh computer, and the number of nuclei were counted using an image analysis software program. There were no significant differences between the number of nuclei counted manually and the number counted using computer-assisted software, until day 7 where the cells were multilayered (P 〈 0.05). This image analysis method was compared to other assays typically used to estimate cell proliferation or population size, namely hemocytometer counting, a rapid colorimetric staining assay using naphthol blue-black, and [3H]-thymidine incorporation. The growth rates derived using image analysis were in close agreement with results derived from hemocytometer counts and [3H]-thymidine incorporation. However, the growth rates of cells grown in high concentrations of FBS as determined using naphthol blue-black were substantially lower than results from image analysis. We conclude that this adaptation of computer-assisted image analysis provides a method to derive accurate growth curves by directly counting the number of cells in a large number of replicates. © 1995 Wiley-Liss, Inc.
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