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  • Microscopy
  • X-ray lasersXFELsbiologystructuredynamics
  • perovskiteferroelectricpowder neutron diffraction
  • protein crystalscrystal lattices
  • Wiley-Blackwell  (9)
  • International Union of Crystallography (IUCr)  (6)
  • Cell Press  (2)
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  • 1
    Publikationsdatum: 2022-05-26
    Beschreibung: © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Hirst, W. G., Kiefer, C., Abdosamadi, M. K., Schäffer, E., & Reber, S. In Vitro reconstitution and imaging of microtubule dynamics by fluorescence and label-free microscopy. STAR Protocols, 1(3), (2020): 100177, doi:10.1016/j.xpro.2020.100177.
    Beschreibung: Dynamic microtubules are essential for many processes in the lives of eukaryotic cells. To study and understand the mechanisms of microtubule dynamics and regulation, in vitro reconstitution with purified components has proven a vital approach. Imaging microtubule dynamics can be instructive for a given species, isoform composition, or biochemical modification. Here, we describe two methods that visualize microtubule dynamics at high speed and high contrast: (1) total internal reflection fluorescence microscopy and (2) label-free interference reflection microscopy.
    Beschreibung: We thank the AMBIO imaging facility (Charité, Berlin) and Nikon at MBL for imaging support. We thank all former and current members of the Reber lab for discussion and helpful advice, in particular Christoph Hentschel and Soma Zsoter for technical assistance. S.R. acknowledges funding by the IRI Life Sciences (Humboldt-Universität zu Berlin, Excellence Initiative/DFG). W.H. was supported by the Alliance Berlin Canberra co-funded by a grant from the Deutsche Forschungsgemeinschaft (DFG) for the International Research Training Group (IRTG) 2290 and the Australian National University. C.K. thanks the Deutsche Forschungsgesellschaft (DFG, JA 2589/1-1). C.K. and M.A. thank Steve Simmert and Tobias Jachowski former and current members of the Schäffer lab.
    Schlagwort(e): Biophysics ; Cell Biology ; Microscopy
    Repository-Name: Woods Hole Open Access Server
    Materialart: Article
    Standort Signatur Erwartet Verfügbarkeit
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  • 2
    Publikationsdatum: 2022-05-26
    Beschreibung: © The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Geisterfer, Z. M., Oakey, J., & Gatlin, J. C. . Microfluidic encapsulation of Xenopus laevis cell-free extracts using hydrogel photolithography. STAR Protocols, 1(3), (2020): 100221, doi:10.1016/j.xpro.2020.100221.
    Beschreibung: Cell-free extract derived from the eggs of the African clawed frog Xenopus laevis is a well-established model system that has been used historically in bulk aliquots. Here, we describe a microfluidic approach for isolating discrete, biologically relevant volumes of cell-free extract, with more expansive and precise control of extract shape compared with extract-oil emulsions. This approach is useful for investigating the mechanics of intracellular processes affected by cell geometry or cytoplasmic volume, including organelle scaling and positioning mechanisms. For complete details on the use and execution of this protocol, please refer to Geisterfer et al. (2020).
    Beschreibung: This work was made possible by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant no. 2P20GM103432. It was also supported by additional funding provided by the NIGMS under grant no. R01GM113028, the NSF Faculty CAREER Program under award no. BBBE 1254608, Whitman Center fellowships at the Marine Biological Laboratory, and the Biomedical Scholars program of the Pew Charitable Trusts. We thank Drs. Aaron Groen and Tim Mitchison for their intellectual contributions and involvement in some of the pioneering experiments that set the foundation for this approach.
    Schlagwort(e): Biophysics ; Cell Biology ; Cell isolation ; Microscopy ; Model Organisms
    Repository-Name: Woods Hole Open Access Server
    Materialart: Article
    Standort Signatur Erwartet Verfügbarkeit
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  • 3
    Digitale Medien
    Digitale Medien
    Weinheim : Wiley-Blackwell
    Journal of High Resolution Chromatography 9 (1986), S. 168-171 
    ISSN: 0935-6304
    Schlagwort(e): Supercritical fluids ; Infrared spectrometry ; Solvent elimination ; Microscopy ; Chemistry ; Analytical Chemistry and Spectroscopy
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Chemie und Pharmazie
    Notizen: The feasibility of supercritical fluid chromatography/Fourier transform-infrared (SFC/FT-IR) microspectrometry is presented. In this approach to SFC/FT-IR, the chromatographic eluates are aspirated from the restrictor directly onto the surface of a moving window which then passes into the beam focus of an infrared microscope. Because the mobile phase is gaseous at ambient conditions, elimination of the mobile phase is easily accomplished. Detection limits in all interfaces between a chromatograph and an FT-IR spectrometer in which the mobile phase is eliminated are determined in large part by the area over which the sample is deposited. We have shown that SFC eluates can be condensed at ambient temperature into spots of 100 to 200μm in diameter. The microscope interface therefore serves to increase the sensitivity of the SFC/FT-IR measurements of these spots and detection limits in the low nanogram range are possible. Preliminary results obtained before any real attempts were made to optimize the deposition process indicate that identifiable spectra can be obtained in real time at the 50 ng level for chromatographic separations performed with a 100μm i.d. wall coated open tubular column. Reproducible reconstructed chromatograms are obtained as each deposited eluate travels through the beam focus of the microscope. The concentration profile of deposited peaks was determined by IR measurements performed at 50 μm increments over the width of the peak to ascertain the deposition size. The results described in this paper, while not yet optimized, indicate the great potential of SFC/FT-IR microspectrometry.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 4
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 29 (1994), S. 344-349 
    ISSN: 1059-910X
    Schlagwort(e): Epithelium ; Eye ; Hyaluronate ; Microscopy ; Rabbit ; Regeneration ; Retina ; Sodium iodate ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: The distribution of hyaluronate (HA) in regenerating retinal pigment epithelium (RPE) of the rabbit was examined using immunohistochemistry and confocal laser scanning microscopy. The goal was to determine if there is a correlation between differentiation and HA expression, like that seen in developing tissues, where HA accumulates and then disappears as the tissue matures. In normal RPE cells HA is associated mainly with the apical surface. In regenerating RPE (produced by i.v. injection of sodium iodate to damage the epithelium, regeneration arising from spared cells), HA exhibits a patchy distribution among the more immature cells and is especially prominent where they overlap or pile up on each other. Where cells are more mature and form a compact monolayer of cells, HA is expressed mainly on the apical surface, as in normal RPE. The accumulation of HA among the more immature cells in the regenerating epithelial sheet supports the hypothesis that HA influences differentiation by suppressing cell-cell associations until the proper time for their formation. © 1994 Wiley-Liss, Inc.
    Zusätzliches Material: 3 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 5
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 21 (1992), S. 171-173 
    ISSN: 1059-910X
    Schlagwort(e): Transmission Electron Microscope ; Light Intensifier Camera ; Microscopy ; Background correction ; Image Analysis ; Video Microscopy ; Television Camera ; Frame Store ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: Several disadvantages of using intensified television cameras to acquire TEM images can be overcome by using background subtraction with a frame store.
    Zusätzliches Material: 1 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 6
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 25 (1993), S. 439-446 
    ISSN: 1059-910X
    Schlagwort(e): Microscopy ; Groundwater ; Pollution ; Radioactive waste ; Transport ; Remediation ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: Environmental colloids are toxic or radioactive particles suspended in ground or surface water. These hazardous particles can facilitate and accelerate the transport of toxicants and enhance the threat to humans by exposure to pathogenic substances. The chemical and physical properties of hazardous colloids have not been well characterized nor are there standard colloid remediation technologies to prevent their deleterious effects. Colloid characterization requires measurement of their size distribution, zeta potential, chemical composition, adsorption capacity, and morphology. The environmental scanning electron microscope (ESEM) by ElectroScan, Inc., analyzes particle sizes, composition, and morphology. It is also used in this study to identify the attachment of colloids onto packing or rock surfaces in our development of a colloid remediation process.The ESEM has confirmed the composition of groundwater colloids in our studies to be generally the same material as the surrounding rock. The morphology studies have generally shown that colloids are simply small pieces of the rock surface that has exfoliated into the surrounding water. However, in general, the source and chemical composition of groundwater colloids is site dependent. We have found that an ESEM works best as a valuable analysis tool within a suite of colloid characterization instruments. © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 7 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 7
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 26 (1993), S. 423-436 
    ISSN: 1059-910X
    Schlagwort(e): Surface forces ; Mucus ; Sol phase ; Airway macrophages ; Microscopy ; Fractionator ; Surface balance ; Video bronchoscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: We have investigated the displacement into the sol phase of inhaled particles deposited in the intrapulmonary conducting airways. Hamsters inhaled an aerosol of monodisperse polystyrene particles of 6 μm diameter. Their lungs were fixed by intravascular perfusion, and light and electron microscopy was used to study the epithelial coating. The surfactant film at the wall-air interface was investigated by measuring its surface tension. The number of particles retained was determined stereologically. In addition we investigated the displacement of spherical particles in vitro on a DPPC monolayer in a Langmuir-Wilhelmy surface balance and determined the surface tension in vivo in the horse trachea by video bronchoscopy, applying the droplet spreading method. We found that particles deposited onto a surfactant film were pulled into the aqueous subphase, and we concluded that surface forces due to the airway surfactant likely displace deposited particles into the periciliary fluid (sol phase). Comparing lungs fixed immediately after inhalation with lungs fixed 24 hr after inhalation revealed that 86% of the particles retained in the intrapulmonary conducting airways immediately after inhalation had been cleared within 24 hr. One-third of the particles of the lungs fixed immediately after inhalation was phagocytized. The combination of structural and stereological analyses with in vitro and in vivo measurements has led to new insights into the role of airway surfactant with respect to the fate of inhaled particles, which may have important consequences regarding the effects of hazardous particles, which may have important consequences regarding the effects of hazardous particles. These studies may also help to evaluate the deposition pattern and clearance of therapeutic particles, with important implications for their therapeutic use. © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 14 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 8
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 23 (1992), S. 252-259 
    ISSN: 1059-910X
    Schlagwort(e): Semiconductor ; Plan view ; Microscopy ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: Photochemical etching (PCE) as a method for preparation of InP semiconductor plan view samples for the transmission electron microscope is demonstrated and compared to the methods of ion milling and chemical thinning. PCE can produce small area samples for TEM analysis quickly and accurately. Also, the resulting thin regions are surrounded by a built-in stabilizing structure that improves handleability and reduces the occurrence of handling induced fracture. © 1992 Wiley-Liss, Inc.
    Zusätzliches Material: 8 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 9
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 27 (1994), S. 83-96 
    ISSN: 1059-910X
    Schlagwort(e): Growth factor ; Inhibin ; Microscopy ; Immunohistochemistry ; In situ hybridization ; Morphology ; Ovary ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: Follicular development in the mammalian ovary is a complex process regulated by an orchestrated action of the pituitary gonadotropins, e.g., follicle stimulating hormone (FSH) and luteinizing hormone (LH), and local ovarian factors, such as peptide growth factors and steroids. The mechanism of endocrine/paracrine/autocrine regulation of folliculogenesis (i.e., cell proliferation and functions) has been addressed largely by biochemical means. However, the availability of immunological and molecular tools now enables us to undertake critical microscopic studies revealing the ovarian cell-type specific synthesis and/or accumulation of many of these local peptide modulators, their roles in the proliferation and differentiation of follicular cells, and their regulation by gonadotropins and local factors. The objective of this review is to provide a comprehensive yet complete picture of the endocrine/autocrine regulation of mammalian folliculogenesis as revealed by microscopic studies. Efforts have been made to include adequate research information relevant to update our understanding of the process of follicular development; however, to maintain the brevity, many equally important studies could not be included. This review confirms that FSH and LH are still the primary stimuli for follicular development. However, it is clear that the actions of these hormones at the cell level involve a host of peptide factors which are produced locally by different follicular cell types and which are powerful modulators of gonadotropin actions. The temporal and spatial expression of the genes of these modulators, the synthesis of active factors, their interactions, and the dynamics of their receptors on the follicular cell surface may be the ultimate determinants of cellular events which are crucial to coordinated growth and differentiation of follicular cells leading to folliculogenesis and ovulation. © 1994 Wiley-Liss, Inc.
    Zusätzliches Material: 6 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 10
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Microscopy Research and Technique 32 (1995), S. 129-147 
    ISSN: 1059-910X
    Schlagwort(e): 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
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: 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.
    Zusätzliches Material: 8 Ill.
    Materialart: Digitale Medien
    Standort Signatur Erwartet Verfügbarkeit
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  • 11
    Digitale Medien
    Digitale Medien
    New York, NY : Wiley-Blackwell
    Journal of Electron Microscopy Technique 11 (1989), S. 137-142 
    ISSN: 0741-0581
    Schlagwort(e): Trachea ; Permeability ; Smoke ; Microscopy ; electron ; Microscopy ; fluorescence ; Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Allgemeine Naturwissenschaft
    Notizen: We studied the penetration of fluorescein isothiocyanate (FITC) T-40 dextran into the paracellular spaces in the tracheal mucosa and its appearance in the blood plasma of guinea pigs exposed to cigarette smoke or (controls) breathing room air. Under general anesthesia, the dextran solution was instilled onto the tracheal surface via a tracheotomy tube, arterial blood was sampled serially for 40 min, and then the trachea was fixed by perfusion or immersion. Examination of the tracheal mucosa with light microscopy, with and without epifluorescence, and transmission electron microscopy, revealed dextran in the paracellular spaces in mucosa from experimental animals but not controls. In all control animals, the levels of the dextran in plasma was below the sensitivity of the assay. By contrast, dextran levels in the plasma from the experimental animals fell within the sensitivity range of the assay and increased in blood at a rate of 0.00125 ± 0.00023 (SE) expressed as a percentage of the instilled dose/min. We conclude that FITC T-40 dextran provides a reliable, fairly simple, fast method for assessing major, but not subtle, changes in paracellular permeability of the tracheal mucosa.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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  • 12
    facet.materialart.
    Unbekannt
    International Union of Crystallography (IUCr)
    In: IUCrJ
    Publikationsdatum: 2017-07-01
    Schlagwort(e): X-ray lasersXFELsbiologystructuredynamics
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
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  • 13
    Publikationsdatum: 2018-05-12
    Schlagwort(e): protein crystalscrystal lattices
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
    Standort Signatur Erwartet Verfügbarkeit
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  • 14
    Publikationsdatum: 2018-05-12
    Schlagwort(e): protein crystalscrystal lattices
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
    Standort Signatur Erwartet Verfügbarkeit
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  • 15
    facet.materialart.
    Unbekannt
    International Union of Crystallography (IUCr)
    In: IUCrJ
    Publikationsdatum: 2017-05-11
    Beschreibung: The development and application of the free-electron X-ray laser (XFEL) to structure and dynamics in biology since its inception in 2009 are reviewed. The research opportunities which result from the ability to outrun most radiation-damage effects are outlined, and some grand challenges are suggested. By avoiding the need to cool samples to minimize damage, the XFEL has permitted atomic resolution imaging of molecular processes on the 100 fs timescale under near-physiological conditions and in the correct thermal bath in which molecular machines operate. Radiation damage, comparisons of XFEL and synchrotron work, single-particle diffraction, fast solution scattering, pump–probe studies on photosensitive proteins, mix-and-inject experiments, caged molecules, pH jump and other reaction-initiation methods, and the study of molecular machines are all discussed. Sample-delivery methods and data-analysis algorithms for the various modes, from serial femtosecond crystallography to fast solution scattering, fluctuation X-ray scattering, mixing jet experiments and single-particle diffraction, are also reviewed.
    Schlagwort(e): X-ray lasersXFELsbiologystructuredynamics
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
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  • 16
    facet.materialart.
    Unbekannt
    International Union of Crystallography (IUCr)
    In: IUCrJ
    Publikationsdatum: 2017-04-27
    Schlagwort(e): perovskiteferroelectricpowder neutron diffraction
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
    Standort Signatur Erwartet Verfügbarkeit
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  • 17
    facet.materialart.
    Unbekannt
    International Union of Crystallography (IUCr)
    In: IUCrJ
    Publikationsdatum: 2017-03-09
    Beschreibung: The perovskite Li0.2Na0.8NbO3 is shown, by powder neutron diffraction, to display a unique sequence of phase transitions at elevated temperature. The ambient temperature polar phase (rhombohedral, space group R3c) transforms via a first-order transition to a polar tetragonal phase (space group P42mc) in the region 150–300°C; these two phases correspond to Glazer tilt systems a−a−a− and a+a+c−, respectively. At 500°C a ferroelectric–paraelectric transition takes place from P42mc to P42/nmc, retaining the a+a+c− tilt. Transformation to a single-tilt system, a0a0c+ (space group P4/mbm), occurs at 750°C, with the final transition to the aristotype cubic phase at 850°C. The P42mc and P42/nmc phases have each been seen only once and twice each, respectively, in perovskite crystallography, in each case in compositions prepared at high pressure.
    Schlagwort(e): perovskiteferroelectricpowder neutron diffraction
    Digitale ISSN: 2052-2525
    Thema: Geologie und Paläontologie , Physik
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