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  • 1
    Electronic Resource
    Electronic Resource
    Preparation of biological tissue sections for correlative ion, electron, and light microscopy (1984)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 1 (1984), S. 299-309 
    Details
    ISSN: 0741-0581
    Keywords: Electron microscopy ; Ion microscopy ; Correlative microscopy ; Electron probe microanalysis ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: In order to correctly interpret the chemical images obtained using ion microscopy (IM), it is useful to correlate them with the information provided by conventional light microscopy (LM), secondary electron imaging (SEI), backscattered electron imaging (BEI), and electron probe microanalysis (EPMA). Accordingly, we have devised a technique of specimen preparation which allows for the application of several different microanalytical techniques to a single histologic section mounted on the same substrate. Sections are cut onto polyester plastic coverslips (devoid of peaks for any element with atomic number 〉 9 using EPMA) and studied by LM. After a light rotary coating with carbon (to prevent charging), the section can then be examined by SEI, BEI, and EPMA. Specific areas can be marked for IM study either with an objective-mounted pin tissue microlocater, or by placing small pieces of metal foil, cut in specific geometric shapes, over features of interest. After sputter-coating the sample with platinum, metal-free shadows are visible using a low-power reflected light microscope available on a typical IM sample chamber as a guide for ion beam placement. The conductive coatings also minimize specimen charging during IM. Post-IM light microscopy, SEI, and BEI are used to confirm the location of specific areas probed in the IM experiments and to provide information on differential ion-sputtering artifacts and tissue contaminants. This new correlative technique should permit better understanding of the images obtained with these diverse instruments.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060010309
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  • 2
    Electronic Resource
    Electronic Resource
    Primary culture of human fallopian tube epithelial cells and co-culture of early mouse pre-embryos (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 32 (1992), S. 236-242 
    Details
    ISSN: 1040-452X
    Keywords: Blastocyst ; Pre-implantation ; EGF ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have established a monolayer culture system for human fallopian tube epithelial cells. The cells were isolated from tubes using collagenase digestion, and were cultured in Ham's F-10 supplemented with 15% fetal calf serum. The epithelial cells derived from culture were characterized using immunocytochemical staining and electron microscopy. These cells were stained with antikeratin and anti-epithelial membrane antigen, but showed no staining after treatment with antivimentin. Electron microscopy showed many microvilli on the cell surface and tight junctions or desmosomes at areas of cell-cell contact. Cell proliferation was enhanced by epidermal growth factor, but not by fibroblast growth factor, insulin, transferrin, estradiol-17β, or progesterone.The 2-cell ICR mouse pre-embryos were co-cultured for 4 days with tubal epithelial cells (A) (n = 98), in cell-conditioned medium (B) (n = 83), or in medium alone (C) (n = 72). During the first 24 h in culture, for groups A and B, the rates of cleavage to the 4-cell stage were 90.9% and 81.9%, respectively. Cleavage rates in these two groups were significantly higher (P =0.0012, P 〉 0.00001) than in group C (56.9%). After 72 h in culture, the rates of development to the blastocyst stage were significantly higher for groups A and B compared to group C (89.6% and 73.5% vs. 54.5%, P 〉 0.00001, P = 0.0002). These results suggest that factor(s) from tubal epithelial cells may facilitate the development of mouse pre-embryos throughout the pre-implantation stages.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080320308
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  • 3
    Electronic Resource
    Electronic Resource
    Localization of bindin expression during sea urchin spermatogenesis (1990)
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 27 (1990), S. 181-190 
    Details
    ISSN: 1040-452X
    Keywords: Testis ; Spermatogenic cells ; Bindin mRNA ; In situ hybridization ; Immunocytochemistry ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Expression of the bindin gene was examined in testicular cells of the sea urchin Strongylocentrotus purpuratus. In situ hybridization studies, using an 35S-labeled antisense RNA probe transcribed from a bindin cDNA, reveal that bindin mRNAs are localized in spermatogenic cells displaced towards the lumens of maturing testicular acini. Little or no hybridization is observed in spermatogenic cells displaced towards the perivisceral epithelium or in somatic cells of the testis. A similar localization of the bindin protein itself is observed using a rhodamine-conjugated polyclonal antibody against bindin, which shows a punctate immunofluorescence pattern in late spermatogenic cells. Immunogold labeling of ultrathin sections and electron microscopy reveal that this punctate immunofluorescence is an apparent result of localized deposits of bindin in intracellular vesicles. Through the terminal stages of spermatogenesis, these bindin-containing vesicles apparently fuse to form the single acrosomal vesicle of the mature spermatozoon. These results indicate (1) that bindin mRNAs are transcribed relatively late in spermatogenesis, (2) that bindin is translated soon after production of its mRNA, (3) that bindin quickly associates with intracellular vesicles during or soon after its synthesis, and (4) that these vesicles fuse to form the single acrosomal vesicle during the terminal stage of spermatogenesis.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1080270302
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  • 4
    Electronic Resource
    Electronic Resource
    Morphological study of the effects of intranasal zinc sulfate irrigation on the mouse olfactory epithelium and olfactory bulb (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 24 (1993), S. 195-213 
    Details
    ISSN: 1059-910X
    Keywords: Deafferentation ; Regeneration ; Sensory afferents ; Electron microscopy ; Olfactory receptor cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The effects of intranasal zinc sulfate (ZnSO4) irrigation on the morphology of the olfactory epithelium and olfactory bulb were studied in mice with short survival times (as early as 1 day) and with long survival times (up to 593 days) after the irrigation procedure. As in several previous studies, the olfactory epithelium was completely destroyed within a few days after the ZnSO4 treatment. Within 2-4 days, the septum and turbinates were covered by a new, cuboidal epithelium, the cells of which differed significantly from any cells normally seen in the olfactory epithelium. Slowly, over several months, small areas of the olfactory epithelium regenerated in many of the animals.The ultrastructural changes occurring in the olfactory bulb from 1 to 25 days (the reactive stage) were characterized by degenerating olfactory axons and axon terminals, hypertrophy of astroglial cell processes, and proliferation of or extravasation by phagocytic cells. By 25 days after intranasal ZnSO4 irrigation, the number of reactive glial processes and phagocytic cells returned to normal. In some mice with survival times of 150 days or longer, there was reinnervation of small areas of the olfactory bulb by regenerated olfactory axons. These new olfactory axons innervated only superficial glomeruli or the outer portions of deeper glomeruli, but they formed synaptic contacts with mitral/tufted cells and periglomerular cells that did not differ from control animals. These findings were supported by tract-tracing experiments with 3H-amino acids and by behavioral analysis.In summary, the ultrastructural changes observed in the olfactory bulb in this study were not significantly different from those observed after surgical lesions of the olfactory epithelium or nerve. The olfactory bulb, however, never fully recovered; glomeruli remained shrunken (though with normal dendro-dendritic synaptic connections), and there was minimal olfactory axon reinnervation. © 1993 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1070240302
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  • 5
    Electronic Resource
    Electronic Resource
    A deposition technique for the imaging and analysis of protein interactions with metal and semiconductor surfaces (1985)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 2 (1985), S. 285-292 
    Details
    ISSN: 0741-0581
    Keywords: Electron microscopy ; Edge-projection TEM ; Field-emission ; Rho protein ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: A new technique for placing biological molecules on metal, insulator, and semiconductor surfaces is described. The procedure requires only 10 μl of solution containing molecules at a concentration of 0.1 - 10 μg/ml. The use of a buffer that does not affect metal substrates, the possibility of fixing the molecules in solution prior to deposition, and the ability to minimize surface tension forces during air drying are other features of the new protocol. Simultaneous deposition on TEM grids and highly curved substrates permits biomolecular adsorption on technologically interesting materials to be visualized in the transmission electron microscope.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060020402
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  • 6
    Electronic Resource
    Electronic Resource
    Utilization of digital image processing to study dynein arms (ATPase) in normal and immotile cilia (1988)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 8 (1988), S. 159-172 
    Details
    ISSN: 0741-0581
    Keywords: Gray scale enhancement ; Immotile cilia syndrome ; Electron microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Tracheal ciliary cross sections were examined with scanning transmission electron microscopy and the resultant images were digitized for image enhancement. A gray-scale histogram of each ciliary image was produced and manipulated to enhance the image for dynein arms. Tracheal epithelial tissue from the pig, rabbit, and dog, including dogs with immotile cilia syndrome, was examined by using this technique. Tissue from each animal was fixed with each of three different fixatives and sections were evaluated for preservation of dynein arms. The same fixative did not consistently provide optimal fixation for ciliary dynein arms in all three species examined. Each species, therefore, must be evaluated to determine the optimal fixative for preservation of normal ciliary ultrastructure. Digital image processing provides a mechanism for enhancing dynein arms in situ without the need for addition of special stains or the use of techniques such as image summation. With this technique it has been shown that about two-thirds of outer dynein arms are partially or completely missing on cilia from dogs with immotile cilia syndrome.
    Additional Material: 12 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060080202
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  • 7
    Electronic Resource
    Electronic Resource
    Structure of superconducting thin films of YBa2Cu3O7-x grown on SrTiO3 and cubic zirconia (1988)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 8 (1988), S. 263-272 
    Details
    ISSN: 0741-0581
    Keywords: High Tc superconductors ; Electron microscopy ; HREM image stimulations ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Thin films of the superconductive oxide YBa2Cu3O7-x have been made by electron-beam coevaporation of the metals in an oxygen atmosphere onto single-crystal {001}-oriented SrTiO3 and yttria-stabilized zirconia (YSZ) substrates. The oxide films were superconducting in the as-deposited state (Tc = 81-83K, Jc = 106 A/cm2 at 4.2K). Bright-field imaging, selectedarea diffraction (SAD), and high-resolution imaging in the transmission electron microscope were used to characterize the microstructure of these films. All of the films were polycrystalline. On SrTiO3 the films were oriented, for the most part, with {110} parallel to the substrate surface. On YSZ, two microstructures were observed: one with smaller rectangular grains oriented with (100) or (010) parallel to the substrate surface and the other with (001) parallel to the surface (i.e., c-axis up).
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060080305
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  • 8
    Electronic Resource
    Electronic Resource
    A Stopped-flow/rapid-freezing machine with millisecond time resolution to prepare intermediates in biochemical reactions for electron microscopy (1990)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 16 (1990), S. 160-166 
    Details
    ISSN: 0741-0581
    Keywords: Stopped-flow ; Rapid-freezing ; Freeze-fracture ; Electron microscopy ; Rapid reactions ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: We have developed an instrument capable of freezing transient intermediates in rapid biochemical reactions for subsequent freeze-fracturing, replication, and viewing by transmission electron microscopy. The machine combines a rapid mixing unit similar to one widely used in chemical kinetics (Johnson, 1986) with a propane jet freezing unit previously used to prepare static samples for freeze-fracturing (Gilkey and Staehelin, 1986). The key element in the system is a unique thin-walled flow cell of copper that allows for injection and aging of the sample, followed by rapid freezing. During freeze-fracturing, a tangential cut is made along the wall of the flow cell to expose the sample for etching and replication. The dead time required for mixing and injection of the reactants into the flow cell is less than 5 ms. Electronic controls allow one to specify, on a millisecond time scale, any time above 5 ms between initiation of the reaction and quenching by rapid freezing.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060160206
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  • 9
    Electronic Resource
    Electronic Resource
    Ion milling of materials science specimens for electron microscopy: A review (1984)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 1 (1984), S. 405-414 
    Details
    ISSN: 0741-0581
    Keywords: Ceramics ; Electron microscopy ; Ion milling ; Specimen preparation ; Sputtering ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: Ion bombardment to perforation is a common technique in the materials sciences by which thin specimens can be prepared for transmission electron microscopy. The process is not without complication and involves radiation damage to the specimen and tends not to preserve the initial specimen topology. Some of the more important facets of the ion-milling process, pertinent to such specimen preparations, are described.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060010409
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  • 10
    Electronic Resource
    Electronic Resource
    Adrenal chromaffin cells as transplants in animal models of parkinson's disease (1989)
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 12 (1989), S. 308-315 
    Details
    ISSN: 0741-0581
    Keywords: Adrenal medulla ; Electron microscopy ; Transplantation ; Plasticity ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Natural Sciences in General
    Notes: The field of neural transplantation has moved rapidly forward in the last decade. Initially, fetal cells were used as implants to investigate their potential to ameliorate deficits in animal models of Parkinson's disease. However, because of the moral and legal problems associated with the use of fetal tissues in humans, alternative sources of donor tissue were sought which possessed the structural and functional characteristics needed to improve motor function in Parkinsonian patients. To date, one of the most promising tissues being investigated is the adrenal medulla, whose chromaffin cells possess an inherent plasticity of form and function. Transplanted chromaffin cells currently are being studied by a variety of approaches, including electron microscopy, in mouse, rat, and primate models of Parkinson's disease. An overview of the role of the chromaffin cell in this exciting and clinically important arena is briefly reviewed, with an emphasis on the fine structure of implanted chromaffin cells.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jemt.1060120403
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