Relevant bibliographies by topics / TIRF

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  4. Book chapters
  5. Conference papers
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Academic literature on the topic 'TIRF'

Author: Grafiati
Published: 4 June 2021
Last updated: 7 July 2024

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Journal articles on the topic "TIRF"

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Treves, Susan, and Francesco Zorzato. "TIRF." Imaging & Microscopy 11, no. 3 (August 2009): 52–53. http://dx.doi.org/10.1002/imic.200990065.

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Truskey, G. A., J. S. Burmeister, E. Grapa, and W. M. Reichert. "Total internal reflection fluorescence microscopy (TIRFM). II. Topographical mapping of relative cell/substratum separation distances." Journal of Cell Science 103, no. 2 (October 1, 1992): 491–99. http://dx.doi.org/10.1242/jcs.103.2.491.

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A simplified model of TIRF optics was used to quantitate the relative membrane/substratum separation distances from the spatial pattern of TIRF image brightness. Phase-contrast and total internal reflection fluorescence microscopy (TIRFM) images were collected of bovine aortic endothelial cells (BAEC) plated onto glass microscope slides for 15 min, 30 min and 24 h. BAEC adherent for 15 min showed an absence of a focal contact morphology, with the region of closest apposition beneath the cell center. After 30 min, multiple contacts with the surface were established and the morphology became more irregular. BAEC attached for 24 h showed well-defined focal contact regions aligned in characteristically striated patterns. The relative distance between closest and farthest membrane/substratum separations are consistent with reported distance between focal and matrix contacts. Topographical maps of membrane/substratum separation distances over the entire ventral surface of the plated cells were constructed to demonstrate the utility of quantitative TIRF microscopy.
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Fu, Yan, Peter W. Winter, Raul Rojas, Victor Wang, Matthew McAuliffe, and George H. Patterson. "Axial superresolution via multiangle TIRF microscopy with sequential imaging and photobleaching." Proceedings of the National Academy of Sciences 113, no. 16 (April 1, 2016): 4368–73. http://dx.doi.org/10.1073/pnas.1516715113.

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We report superresolution optical sectioning using a multiangle total internal reflection fluorescence (TIRF) microscope. TIRF images were constructed from several layers within a normal TIRF excitation zone by sequentially imaging and photobleaching the fluorescent molecules. The depth of the evanescent wave at different layers was altered by tuning the excitation light incident angle. The angle was tuned from the highest (the smallest TIRF depth) toward the critical angle (the largest TIRF depth) to preferentially photobleach fluorescence from the lower layers and allow straightforward observation of deeper structures without masking by the brighter signals closer to the coverglass. Reconstruction of the TIRF images enabled 3D imaging of biological samples with 20-nm axial resolution. Two-color imaging of epidermal growth factor (EGF) ligand and clathrin revealed the dynamics of EGF-activated clathrin-mediated endocytosis during internalization. Furthermore, Bayesian analysis of images collected during the photobleaching step of each plane enabled lateral superresolution (<100 nm) within each of the sections.
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Foylan, S., W. B. Amos, J. Dempster, L. Kölln, C. G. Hansen, M. Shaw, and G. McConnell. "MesoTIRF: A prism-based Total Internal Reflection Fluorescence illuminator for high resolution, high contrast imaging of large cell populations." Applied Physics Letters 122, no. 11 (March 13, 2023): 113701. http://dx.doi.org/10.1063/5.0133032.

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Total internal reflection fluorescence (TIRF) illumination bypasses the axial diffraction limit of light by using an evanescent field to excite fluorophores close to a sample substrate. However, standard TIRF imaging through the objective requires a high numerical aperture (NA) to generate the evanescent wave. Available lenses have a high magnification with a correspondingly small field of view—ranging from [Formula: see text]50 μm to 1 mm in diameter. Switching to the older prism-TIRF configuration introduced by Axelrod in the 1980s might seem to remove the requirement for high objective NA and allow the use of existing large-field objectives. Unfortunately, these lenses are unsuitable because their throughput of light is too low for TIRF imaging. As such, high sensitivity TIRF imaging over a much larger mesoscopic field has yet to be demonstrated. We have developed a prism-based TIRF illuminator for the Mesolens—a highly corrected objective lens with an unparalleled ratio of NA to magnification. The imaging field of the Mesolens is 204 times larger than that of the TIRF objectives previously described, increasing the optical throughput of the optical system by a factor of 25 compared to an off-the-shelf microscope objective of the same magnification. We demonstrate MesoTIRF imaging of cell specimens and show the multi-wavelength capability of the modality across more than 700 cells in a single image.
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Lin, Jia, and Adam D. Hoppe. "Uniform Total Internal Reflection Fluorescence Illumination Enables Live Cell Fluorescence Resonance Energy Transfer Microscopy." Microscopy and Microanalysis 19, no. 2 (March 11, 2013): 350–59. http://dx.doi.org/10.1017/s1431927612014420.

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AbstractFluorescence resonance energy transfer (FRET) microscopy is a powerful technique to quantify dynamic protein-protein interactions in live cells. Total internal reflection fluorescence (TIRF) microscopy can selectively excite molecules within about 150 nm of the glass-cell interface. Recently, these two approaches were combined to enable high-resolution FRET imaging on the adherent surface of living cells. Here, we show that interference fringing of the coherent laser excitation used in TIRF creates lateral heterogeneities that impair quantitative TIRF-FRET measurements. We overcome this limitation by using a two-dimensional scan head to rotate laser beams for donor and acceptor excitation around the back focal plane of a high numerical aperture objective. By setting different radii for the circles traced out by each laser in the back focal plane, the penetration depth was corrected for different wavelengths. These modifications quell spatial variations in illumination and permit calibration for quantitative TIRF-FRET microscopy. The capability of TIRF-FRET was demonstrated by imaging assembled cyan and yellow fluorescent protein–tagged HIV-Gag molecules in single virions on the surfaces of living cells. These interactions are shown to be distinct from crowding of HIV-Gag in lipid rafts.
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Lanni, F., A. S. Waggoner, and D. L. Taylor. "Structural organization of interphase 3T3 fibroblasts studied by total internal reflection fluorescence microscopy." Journal of Cell Biology 100, no. 4 (April 1, 1985): 1091–102. http://dx.doi.org/10.1083/jcb.100.4.1091.

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We studied the laminar organization of 3T3 fibroblast cells growing on glass slides by use of total internal reflection illumination to excite fluorescence emission (TIRF) from labeled molecules and stained cellular compartments that are very close to the cell-substrate contact region. Mitochondria, distant from the contact regions and stained with the water-soluble cationic dye, dil-C3-(3), fluoresced only as the glass/cytoplasm critical angle was approached. A similar result was obtained when the nuclei were stained with Hoechst dye 33342. From this measured angle a cytoplasmic refractive index in the range 1.358-1.374 was computed. The plasma membrane of 3T3 cells was stained with dil-C18-(3), and the cytoplasmic compartment was stained with fluoresceinyl-dextran (FTC-dextran) or with carboxyfluorescein. We have demonstrated a high degree of correspondence between the low-reflectance zones in the reflection interference image of a live cell and the TIRF images of both the plasma membrane and cytoplasmic compartment. TIRF photometry of selected contact regions of cells provided data from which the absolute separation of cell and substrate was computed. From a population of 3T3 cells microinjected with fluorescein-labeled actin, motile and adherent interphase cells were selected for study. For adherent cells, which displayed fluorescent stress fibers, the TIRF image was composed of intense patches and less intense regions that corresponded, respectively, to the focal contact and close-contact zones of the reflection-interference image. The intense patches corresponded to the endpoints of the stress fibers. Cells of motile morphology, which formed some focal contacts and extensive close-contact zones, gave AF-actin TIRF images of relatively even intensity. Thin lamellar regions of the cytoplasm were found to contain concentrations of actin not significantly different from other close-contact regions of the cell. The major analytical problem of TIRF microscopy is separation of the effects of proximity to substrate, refractive index, and fluorescent probe concentration on the local brightness of the TIRF image. From our results, it appears possible to use TIRF microscopy to measure the proximity of different components of substrate contact regions of cells.
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Morelo Pereira, Douglas Jahir, and Diocelina Torres Castro. "Técnicas e indicadores de rendimiento financiero aplicados al estado de resultados en empresas comerciales y de servicios colombianas." Cuadernos de Contabilidad 22 (August 19, 2021): 1–21. http://dx.doi.org/10.11144/javeriana.cc22.tirf.

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Este artículo está dirigido a identificar el grado de utilización de un conjunto de Técnicas e Indicadores de Rendimiento Financiero (TIRF) que permitan monitorear la sostenibilidad financiera de las empresas comerciales y de servicios. El problema radica en las frecuentes reformas tributarias y la constante lucha contra el contrabando; actividades que en su conjunto conjugan una amenaza a la sostenibilidad financiera de la empresa. En este sentido, la presente investigación busca analizar las características predominantes, diferenciales y correlacionales de las TIRF asociadas al estado de resultados. El estudio se desarrolló bajo el paradigma cuantitativo lo que permitió aplicar un cuestionario de nuevo diseño, la muestra estuvo conformada por 90 expertos con funciones asociadas al análisis de estados financieros vinculados a 55 empresas comerciales y 35 empresas de servicios seleccionadas de un listado de 1.000. Entre las conclusiones se determinó que las empresas de servicios tienden a realizar más proyecciones de tendencia y benchmarking contra un competidor clave que las empresas de comerciales. Situación que permite inferir la versatilidad y el dinamismo de estas TIRF en el análisis de estados financieros.
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Gingell, D., O. S. Heavens, and J. S. Mellor. "General electromagnetic theory of total internal reflection fluorescence: the quantitative basis for mapping cell-substratum topography." Journal of Cell Science 87, no. 5 (June 1, 1987): 677–93. http://dx.doi.org/10.1242/jcs.87.5.677.

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Total internal reflection fluorescence (TIRF) has recently been used to look at the contacts made between cells and a glass surface on which they are spread. Our method utilizes the fluorescence of a water-soluble dye that acts as an extracellular aqueous volume marker. Fluorescence is stimulated by the short-range electric field near the glass surface that exists under conditions of total internal reflection. Since fluorescence is normally generated beneath a spread cell and not beyond it, the fluorescence of the image is related to the size of the cell-glass water gap. The images obtained are remarkable for their detail, contrast and the absence of confusing granularity due to cytoplasmic heterogeneity, which is commonly seen in interference reflection (IRM) images. We here develop a rigorous electromagnetic theory of total internal reflection in layered structures appropriate for cell contacts and apply it to quantitative TIRF. We show that: (1) TIRF, unlike IRM, can report cell-glass gaps in a way that is practically independent of the detailed physical properties of the cell; (2) TIRF is also far more sensitive than IRM for measuring cell-glass water gaps up to approximately equal to 100nm. These striking results explain the image quality seen by TIRF. As the initial step towards verifying our theory we show that measurement of the fluorescence stimulated by total internal reflection at a simple glass-water interface matches theoretical predictions.
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Ellefsen, Kyle L., Joseph L. Dynes, and Ian Parker. "Spinning-Spot Shadowless TIRF Microscopy." PLOS ONE 10, no. 8 (August 26, 2015): e0136055. http://dx.doi.org/10.1371/journal.pone.0136055.

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Balaa, Karla, Emmanuel Fort, and Nikon Instruments. "Surface Plasmon Enhanced TIRF Imaging." Imaging & Microscopy 11, no. 4 (October 30, 2009): 55–56. http://dx.doi.org/10.1002/imic.200990091.

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Dissertations / Theses on the topic "TIRF"

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Gortari, Antu Nehuen. "Metasurfaces for bioimaging." Thesis, Université Paris-Saclay (ComUE), 2019. http://www.theses.fr/2019SACLS416/document.

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Au cours des dernières années, des efforts importants ont été déployés pour développer des métasurfaces (MSs) électromagnétiques avec la possibilité de changer de manière abrupte les propriétés de la lumière. Ces avancées ont ouvert une nouvelle gamme de possibilités pour contrôler la lumière en utilisant des dispositifs optiques ultra-minces. Dans ce contexte, et plus spécifiquement dans le spectre visible, les applications en bio-imagerie s’avèrent particulièrement intéressantes. Une technique qui est particulièrement bien adaptée à l'étude de molécules proches d'une membrane cellulaire est la microscopie à fluorescence par réflexion interne (TIRFM), qui repose sur un champ évanescent d'excitation. Dans ce cas la lumière incidente est totalement réfléchie sur une interphase (typiquement verre/eau) en raison de son angle d'incidence élevé. À ce jour, la TIRFM est généralement mise en œuvre à l'aide d'objectifs volumineux de grande ouverture numérique et de petit champ de vision.Dans ce travail de thèse, nous réalisons de substrats pour la microscopie TIRF à base de métasurfaces constituées de réseaux périodiques de structures asymétriques fabriquées en dioxyde de titane (TiO2) sur du verre borosilicaté. Ces structures, aussi petites que 48 nm, ont été optimisées à l’aide de simulations numériques "Rigorous coupled-wave analysis” (RCWA) dans le but de coupler de 50 à 90% de la lumière incidente dans le premier ordre de diffraction avec des angles élevés (θ > 63deg). Le fait de pouvoir utiliser des objectifs de faible grossissement et d'avoir une grande zone de champ évanescent fournit des conditions TIRF uniques qui ne sont pas accessibles par les méthodes traditionnelles. De plus, ces structures sont compatibles avec la lithographie par nanoimpression UV, ce qui permet d’envisager une fabrication à bas coût et à grande échelle. Outre la conception, et la fabrication, dans cette thèse nous aboutissons à une preuve de principe de la microscopie TIRF basée sur des métasurfaces en milieu biologique en imageant notamment des membranes fluorescentes de cellules souches. Ces métasurfaces permettent ainsi l’implémentation TIRFM à contraste élevé et à faible photo-blanchissement compatible avec des microscopes à champ large peu coûteux
In recent years there has been a significant effort to push electromagnetic metasurfaces with the ability to abruptly change light properties into visible wavelengths. These advancements have opened a new range of possibilities to reshape light using ultra-thin optical devices and there is one field that is starting to gather attention: bioimaging. One technique particularly well suited for the study of molecules near a cell membrane is Total Internal Reflection Fluorescence (TIRF) microscopy, which relies on an evanescence field created by light being totally internally reflected within a glass substrate due to its high incidence angle. As of today, TIRF is generally implemented using bulky high-NA, small field of view oil objectives.In this project we present the realization of metasurface-based TIRF microscopy substrates consisting of periodic 2D arrays of asymmetric structures fabricated in titanium dioxide on borosilicate glass. These patterns, as small as 48nm, were optimized through rigorous coupled-wave analysis to couple 50-90% of the incoming normally incident light into the first diffraction order, which outputs at an angle that suffices total internal reflection in water and eliminates the requirement for high NA objectives or prisms to achieve TIRF. Being able to utilize lower-magnification air objectives and having a large evanescence field area provide unique TIRF conditions not accessible by traditional methods. Additionally, these structures are compatible with soft UV nanoimprint lithography, for cost-effective scale production, to give TIRF’s high contrast, low photodamage and low photobleaching capabilities to inexpensive wide-field microscopes
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Burgess, Helen Jane. "A bioelectrochemical FRET switch : combined total internal reflection (TIRF) microscopy and electrochemistry." Thesis, University of Oxford, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.437177.

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Uhlig, Katja. "Untersuchungen PEG-basierter thermo-responsiver Polymeroberflächen zur Steuerung der Zelladhäsion." Phd thesis, Universität Potsdam, 2010. http://opus.kobv.de/ubp/volltexte/2010/4778/.

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Moderne Methoden für die Einzelzellanalyse werden dank der fortschreitenden Weiterentwicklung immer sensitiver. Dabei steigen jedoch auch die Anforderungen an das Probenmaterial. Viele Aufbereitungsprotokolle adhärenter Zellen beinhalten eine enzymatische Spaltung der Oberflächenproteine, um die Ablösung vom Zellkultursubstrat zu ermöglichen. Verschiedene Methoden, wie die Patch-Clamp-Technik oder eine auf der Markierung extrazellulärer Domänen von Membranproteinen basierende Durchflusszytometrie können dann nur noch eingeschränkt eingesetzt werden. Daher ist die Etablierung neuer Zellablösemethoden dringend notwendig. In der vorliegenden Arbeit werden erstmals PEG-basierte thermo-responsive Oberflächen erfolgreich für die Zellkultur eingesetzt. Dabei wird das zerstörungsfreie Ablösen verschiedener Zelllinien von den Oberflächen durch Temperatursenkung realisiert. Die Funktionalität der Oberflächen wird durch Variation der Polymerstruktur, sowie der Konzentration der Beschichtungslösung, durch Beschichtung der Oberflächen mit einem zelladhäsionsfördernden Protein (Fibronektin) und durch Adsorption zelladhäsionsvermittelnder Peptide (RGD) optimiert. Um den Zellablösungsprozess detaillierter zu untersuchen, wird hier zum ersten Mal der direkte Zellkontakt mit thermo-responsiven Oberflächen mittels oberflächensensitiver Mikroskopie (TIRAF) sichtbar gemacht. Mit dieser Technik sind die exakte Quantifizierung und die Analyse der Reduktion der Zelladhäsionsfläche während des Abkühlens möglich. Hierbei werden in Abhängigkeit von der Zelllinie Unterschiede im Zellverhalten während des Ablösens festgestellt: Zellen, wie eine Brustkrebszelllinie und eine Ovarzelllinie, die bekanntermaßen stärker mit ihrer Umgebung in Kontakt treten, vergrößern im Verlauf des Beobachtungszeitraumes den Abstand zwischen Zellmembran und Oberfläche, reduzieren jedoch ihre Zell-Substratkontaktfläche kaum. Mausfibroblasten hingegen verkleinern drastisch die Zelladhäsionsfläche. Der Ablösungsprozess wird vermutlich aktiv von den Zellen gesteuert. Diese Annahme wird durch zwei Beobachtungen gestützt: Erstens verläuft die Reduktion der Zelladhäsionsfläche bei Einschränkung des Zellmetabolismus durch eine Temperatursenkung auf 4 °C verzögert. Zweitens hinterlassen die Zellen Spuren, die nach dem Ablösen der Zellen auf den Oberflächen zurückbleiben. Mittels Kombination von TIRAF- und TIRF-Mikroskopie werden die Zelladhäsionsfläche und die Aktinstruktur gleichzeitig beobachtet. Die Verknüpfung beider Methoden stellt eine neue Möglichkeit dar, intrazelluläre Prozesse mit der Zellablösung von thermo-responsiven Oberflächen zu korrelieren.
Modern methods for single-cell analysis are becoming increasingly sensitive. At the same time, requirements for the sample material are on the rise. Today, sample preparation of adherent cells usually includes steps of enzymatic treatment to digest surface proteins thus, inducing cell detachment from culture substrates. This strongly limits the application of different techniques like patch clamp or labelling of extracellular domains of membrane proteins for flow cytometry. Therefore, a new cell detachment method is urgently required. In the present work, new PEG-based thermo-responsive polymers are used for cell culture for the first time. Here, non-destructive detachment of different cell lines from polymer-coated surfaces is realised by controlled temperature reduction. The surface functionality is systematically optimised by varying the concentration of the coating solutions, by artificial surface coating of a cell adhesion-mediating protein (fibronectin) and by co-adsorption of a cell adhesion-mediating peptide (RGD). For detailed analysis of the cell detachment process, TIRF microscopy is used to directly visualise the cell contacts on the thermo-responsive surfaces. Using this technique allows both the quantification and analysis of the reduction of the cell adhesion area during sample cooling. Furthermore, for several cell lines, different behaviours in cell detachment are observed. Cells that have close contact to their substrate like MCF-7 breast cancer cell line and CHO-K1 ovary cells increase the distance between cell membrane and surface, but there is only little decrease of cell-substrate adhesion area. In contrast, L929 fibroblasts reduce the cell adhesion area drastically. Furthermore, the hypothesis that the cell detachment is an active process is shown by lowering the cell metabolism by temperature reduction to 4 °C and by the cell traces that are left behind after rinsing the surfaces. A combination of TIRAF and TIRF enables visualising the cell adhesion area and actin structures. Measuring both parameters simultaneously opens up new possibilities to correlate intracellular and cell detachment processes on thermo-responsive surfaces.
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Ries, Jonas. "Advanced Fluorescence Correlation Techniques to Study Membrane Dynamics." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2008. http://nbn-resolving.de/urn:nbn:de:bsz:14-ds-1219846317196-73420.

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Fluorescence Correlation Spectroscopy (FCS) is a powerful tool to measure important physical quantities such as concentrations, diffusion coefficients, diffusion modes or binding parameters, both in solution and in membranes. However, it can suffer from severe artifacts, especially in non-ideal systems. Here we develop several novel implementations of FCS which overcome these limitations and facilitate accurate and quantitative determination of dynamic parameters in membranes. Two-focus FCS with camera-detection allows for accurate and calibration-free determination of diffusion coefficients. Confocal FCS using a laser scanning microscope provides an unprecedented positioning accuracy which enabled us to study, for the first time with FCS, dynamics in bacterial membranes. Scanning FCS with a scan path perpendicular to the membrane plane allows to correct for instabilities permitting long measurement times necessary to study slow diffusion. It can easily be extended to measure calibration-free diffusion coefficients with two-focus scanning FCS and to quantify binding with dual color scanning FCS. Spectral crosstalk can be avoided effectively by using alternating excitation. Using this method we were able to perform measurements in systems previously not accessible with FCS, such as yeast cell membranes or membranes of living zebrafish embryos. Line-scan FCS with a scan path in the membrane plane uses the parallel acquisition along the line to increase the statistical accuracy and decrease the measurement times. Knowledge of the scan speed serves as an internal calibration, enabling accurate diffusion and concentration measurements within seconds, hardly affected by photobleaching. Both realizations of scanning FCS can be easily implemented with commercial laser scanning microscopes. Often, a fluorescence background around the membrane cannot be avoided. The high surface selectivity needed in this case can be achieved efficiently by using a novel objective for FCS, the supercritical angle objective, which produces a very flat and laterally confined detection volume. Another technique with similar surface selectivity is FCS with total internal reflection excitation (TIRFCS). Due to the lack of a correct model, the accurate analysis of TIR-FCS data was previously not possible. In this work we develop such a model, enabling quantitative measurements of membrane dynamics with TIR-FCS. The novel FCS techniques developed here will have a high impact on the use of FCS to address key questions in biological systems, previously inaccessible by other methods
Fluoreszenz-Korrelations-Spektroskopie (FCS) ist eine mächtige Methode, um wichtige physikalische Parameter wie Konzentrationen, Diffusionskoeffizienten, Diffusionsarten oder Bindungsparameter in Lösung und in Modell- oder Zellmembranen zu bestimmen. In nichtidealen Systemen ist FCS fehleranfällig. In dieser Arbeit entwickeln wir mehrere neuartige Realisierungen von FCS, welche diese Fehlerquellen umgehen und die genaue und quantitative Messung dynamischer Parameter in Membranen ermöglichen. Zwei-Fokus FCS mit Kamera-Detektion erlaubt eine genaue und kalibrationsfreie Messung von Diffusionskoeffizienten. Konfokale FCS mit einem Laserscanningmikroskop besitzt eine bislang unerreichte Positionsgenauigkeit, welche uns erstmals dynamische Messungen in Bakterienmembranen mit FCS ermöglichte. Scanning FCS mit einem Scanweg senkrecht zur Membran ermöglicht eine Korrektur von Instabilitäten und damit lange Messzeiten, die zur Bestimmung langsamer Diffusionskoeffizienten notwendig sind. Eine Erweiterung zur kalibrationsfreien Messung von Diffusionskoeffizienten mit Zwei-Fokus Scanning FCS und von Bindungsparametern mit Zwei-Farben Scanning FCS ist einfach. Mit diesen Methoden konnten wir in Systemen messen, die bislang FCS nicht zugänglich waren, so in Hefezellmembranen oder in Membranen lebender Zebrafischembryonen. Line-scan FCS besitzt einen Scanweg parallel zur Membran. Die parallele Messung entlang der ganzen Linie führt zu einer deutlichen Verbesserung der Statistik und damit zu kurzen Messzeiten. Die Kenntnis der Scangeschwindigkeit dient einer internen Kalibration und erlaubt eine akkurate Bestimmung von Diffusionskoeffizienten und Konzentrationen innerhalb weniger Sekunden, kaum beeinflusst vom Bleichen von Fluorophoren. Beide Arten von Scanning FCS können mit einem kommerziellen Laserscanningmikroskop realisiert werden. Häufig kann bei FCS Messungen ein fluoreszierender Hintergrund nicht vermieden werden. Hier ist eine hohe Oberflächenselektivitiät nötig, welche effizient mit einem neuartigen Objektiv erreicht werden kann. Dieses Supercritical Angle-Objektiv erzeugt ein sehr flaches und lateral begrenztes Detektionsvolumen. Eine weitere Methode mit einer ähnlich guten Oberflächenselektivität ist FCS mit Anregung über totale interne Reflektion (TIR-FCS). Bislang war eine quantitative Analyse der TIR-FCS Daten kaum möglich, da keine ausreichend genaue theoretische Beschreibung existierte. In dieser Arbeit entwickeln wir ein akkurates Modell, welches quantitative Messungen mit TIR-FCS erlaubt. Die hier entwickelten neuartgien FCS-Techniken ermöglichen die Untersuchung biologischer Fragestellungen, welche bislang keiner anderen Methode zugänglich sind
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Nathwani, Bhavik Bharat. "Developing luminescent nanoprobes for labeling focal adhesion complex proteins and performing combined AFM-TIRF imaging of these conjugates." Texas A&M University, 2008. http://hdl.handle.net/1969.1/85904.

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Recent progress in the field of semiconductor nanocrystals or Quantum Dots (QDs) has seen them find wider acceptance as a tool in biomedical research labs. As produced, high quality QDs synthesized by high temperature organometallic synthesis, are coated with a hydrophobic ligand. Therefore, they must be further processed to be soluble in water and made biocompatible. A process to coat the QDs with silk fibroin, a fibrous protein derived from the Bombyx mori silk worm, is described. Following the coating process, the characterization of size, optical properties and biocompatibility profile of these particle systems is described. In addition, conjugation of the silk fibroin coated QDs to different labeling proteins such as phalloidin and streptavidin is described. Proteins on the surface of ovarian cancer cells (HeyA8) and of cytoskeletal components participating in the formation of focal adhesion complex (FAC), such as F-actin in endothelial cells (HUVECS) were labeled using the bio-conjugated QDs. Various imaging techniques such as epi-fluorescence, TIRF and AFM were used to study the QD labeled cells. Overall the project has produced luminescent nanoprobes that enable the study of FAC formation dynamics and potentially a better in vivo fluorescent marker tool.
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Bethea, Tomika R. C. "Silica Colloidal Crystals as Porous Substrates for Total Internal Reflection Fluorescence Microscopy." Thesis, The University of Arizona, 2006. http://hdl.handle.net/10150/193371.

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In cell biology and chemistry, total internal reflection microscopy (TIRFM) has proven to be a useful technique that allows the probing of cellular processes with high-signal-to-noise ratio imaging. However, samples on solid substrates limit the accessibility to probe processes on extracellular membrane surface closest to the microscope objective. Colloidal crystals provide a porous alternative to the traditional solid substrates. Thin crystals exhibit optical properties similar to that of a fused silica coverslip allowing for TIRFM in the same manner as with a typical coverslip as demonstrated by the observance of Chinese hamster ovary cells with fluorescently labeled receptors on both types of substrates. Accessibility of the cell membrane closest to the substrate and the ability to probe fluorophore orientation information was observed by the binding of TIPP-cy5 to the human delta opioid receptor.
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Tassinari, Andrea 1991. "In vitro reconstitution of the collective activities of motor protein complexes." Doctoral thesis, Universitat Pompeu Fabra, 2020. http://hdl.handle.net/10803/670056.

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Tschmelak, Jens M. [Verfasser]. "New ultra-sensitive immunoassays for (Total Internal Reflectance Fluorescence) TIRF-based biosensors / Jens M Tschmelak." Aachen : Shaker, 2005. http://d-nb.info/1186578238/34.

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Faure, Laura. "La machinerie de motilité de Myxococcus xanthus : caractérisation d'une nouvelle famille de moteurs moléculaires dans l'enveloppe bactérienne." Thesis, Aix-Marseille, 2017. http://www.theses.fr/2017AIXM0011/document.

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Dans les cellules il existe deux grandes sources d’énergie : l’ATP et la force proton-motrice, produites au niveau du cytoplasme et de la membrane interne respectivement. La mise en place de processus actifs dans la membrane externe ou à la surface des bactéries à Gram négatif requière la présence de machineries protéiques transmettant les forces de leur lieu de production à leur lieu d’utilisation. Durant ma thèse j’ai étudié une de ces machines : la machinerie de motilité (Agl-Glt) de Myxococcus xanthus. Plus précisément, j’ai cherché à comprendre comment les composants de cette machine s’organisent pour permettre le déplacement d’une bactérie. J’ai montré que l’assemblage de la machinerie de motilité au pôle avant des cellules nécessite la formation d’une plateforme cytosolique sur laquelle vient se fixer la machine Agl-Glt. Sous l’action du moteur, le complexe interne de la machine se déplace en direction du pôle arrière en suivant une trajectoire hélicoïdale de main droite. Au niveau de la surface les protéines de membrane externe sont recrutées au niveau d’adhésions focales et permettent l’ancrage de la machinerie au substrat. Enfin, la transmission des forces de la membrane interne à la surface par la machinerie de motilité génère le déplacement des cellules selon une trajectoire hélicoïdale de main gauche. Finalement, cette étude a révélé l’existence d’une machine protéique de l’enveloppe dont l’activité repose sur l’association d’un moteur linéaire et du cytosquelette bactérien. De par l’homologie qu’il existe entre les systèmes il est possible de proposer que ce type de machines peut-être retrouvé associées à d’autres fonctions que la motilité cellulaire
Two energy sources are present in cells: the ATP and the Proton Motive Force, produced in the cytoplasm and inner membrane respectively. Active processes in the outer membrane or on the surface of Gram negative bacteria require the presence of a proteic machinery to transduce the forces from their production site, in the cytoplasm or inner membrane, to their usage site. During my thesis I have studied one of these machineries: the motility machinery (Agl-Glt) of Myxococcus xanthus. More precisely, I try to understand how the components of this transmembrane machinery interact with each other to promote cell motility. I have shown that the assembly of the motility machinery at the leading pole requires the formation of a cytoplasmic platform onto which the Agl-Glt machinery is going to nucleate. The inner-membrane motor complex moves intracellularly along a right-handed path in the cell and becomes stationary at focal adhesion sites on the surface through the connection of the motor to the outer membrane proteins of the complex. This powers the left-handed helical motion of the bacteria. Finally, this study reveals the existence of a dynamic transmembrane machinery which associates the bacterial cytoskeleton to a linear motor to promote cell movement. The homology between the systems tells us that this type of motor is likely to be found associate with other function than cell motility
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Nassereddine, Aya. "Surface patterning strategies to dissect T-Cell adhesion and actin organisation." Thesis, Aix-Marseille, 2019. http://www.theses.fr/2019AIXM0458.

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Pour une réponse immunitaire efficace, une interaction optimale entre les cellules T et les cellules présentatrices d'antigène (APC) est nécessaire. Elle se présente sous la forme d'un contact cellulaire à différentes échelles allant du moléculaire (1-10 nm) au cellulaire (1-10 micromètres). La liaison entre les récepteurs spéciaux des cellules T (TCR) et leurs ligands sur une APC entraîne une réorganisation moléculaire à plus grande échelle menant d'abord à la formation de micro-clusters de TCR, puis à une restructuration à l'échelle cellulaire de la membrane et du cytosquelette. La création d'un substrat artificiel avec des amas de ligands qui induisent à leur tour l'accumulation de TCR est un outil important pour comprendre le lien entre l'organisation du TCR et de son ligand, l'organisation du cytosquelette d'actine et l'impact de ces deux facteurs sur le comportement cellulaire global, notamment l'adhérence et la signalisation. Nous avons développé un nouveau substrat basé sur la nanotechnologie et utilisé une stratégie alternative basée sur l'auto-assemblage colloïdal pour montrer que le TCR est clairement groupé sur des points de 700 nm et non pas sur des points de 400 nm. L'actine est distribuée de manière homogène sous forme de réseau dans la plupart des cellules, mais dans quelques-unes d'entre elles, elle apparaît sous la forme de points co-localisés avec les amas de ligands. Une observation plus fine à l'aide de la microscopie de reconstruction optique aléatoire indique que les points peuvent en fait être des sites où des faisceaux d'actine se croisent pour former des nœuds non visibles à moindre résolution
For an efficient immune response, an optimal interaction between T-cells and antigen presenting cells (APC) is required; it takes the form of a cell-cell contact involving different scales ranging from the molecular (1-10 nm) to the cellular (1-10 micrometer). The ligation of the special T cell receptors (TCR) to its ligands on an APC, leads to larger scale molecular reorganisation leading first to formation of TCR micro-clusters, and later to cell-scale restructuring of both the membrane and the cytoskeleton. Patterning an artificial substrate with ligand-clusters that in turn induce TCR-clustering is an important tool to understand the link between the organisation of TCR and its ligand, the organisation of the actin cytoskeleton and the impact of both on overall cell behavior including adhesion and signaling. We developed a new nanotechnology based substrate (ligand-dot size down to 250 nm) and also used an alternative strategy based on colloidal self-assembly (700 or 400 nm) to show that TCR is clearly clustered on 700 nm dots but not on smaller 400 nm dots. Actin is homogeneously distributed in the form of a network in most cells but in a few of them, it appears as dots that co-localize with the ligand clusters. Finer observation using stochastic optical reconstruction microscopy indicates that the dots may in fact be sites where actin bundles cross each other forming nodes that are not visible at lower resolution. This work confirms a close link between T cell receptor organisation and actin structure
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Books on the topic "TIRF"

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INSA NEW-ICTE '96 (1996 New Delhi, India). Information technology in education and research: Proceedings of INSA NEW-ICTE '96 (INSA, New Delhi) and UNESCO-CICT Workshop (TIRF, Mumbai). Edited by Govil Rekha, Singhi Navin M, Indian National Science Academy, and UNESCO-CICT Workshop (1997 : Bombay, India). New Delhi: Indian National Science Academy, 1998.

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Claeson, Stewe. Tiro. Stockholm: Norstedt, 2007.

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Giapponi, Thomas. Tire Forensic Investigation Analyzing Tire Failure. Warrendale, PA: SAE International, 2008. http://dx.doi.org/10.4271/r-387.

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Tire forensic investigation: Analyzing tire failure. Warrendale, Pa: SAE International, 2009.

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Gurusami, M. Rakkappa Pothilinga. Tiru. Vi. Ka.: Tiru. Vi. Kaliyāṇacuntara Mutaliyār. New Delhi: Cākittiya Akkātemi, 1998.

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Tiff gear: The autobiography of Tiff Needell. Newbury Park, CA: Haynes Publishing, 2012.

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Tiff gear: The autobiography of Tiff Needell. Sparkford, Yeovil, Somerset: Haynes, 2011.

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Tiro directo. Santafé de Bogotá: Planeta Colombiana Editorial, 1998.

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Cheng, Andrea. Tire mountain. Honesdale, Pa: Boyds Mills Press, 2007.

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Wahyudi, Aris. Tuhan tiri. [Jakarta]: Voxdei Publications, 2003.

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Book chapters on the topic "TIRF"

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Taylor, Donald. "TIRF REMS Access Program." In Managing Cancer Breakthrough Pain, 89–92. Tarporley: Springer Healthcare Ltd., 2013. http://dx.doi.org/10.1007/978-1-908517-83-8_6.

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Sapsford, Kim E., and Frances S. Ligler. "TIRF Array Biosensor for Environmental Monitoring." In Optical Sensors, 359–90. Berlin, Heidelberg: Springer Berlin Heidelberg, 2004. http://dx.doi.org/10.1007/978-3-662-09111-1_14.

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Grawenhoff, Julia, Sebastian Baumann, and Sebastian P. Maurer. "In Vitro Reconstitution of Kinesin-Based, Axonal mRNA Transport." In Methods in Molecular Biology, 547–68. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-1990-2_29.

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AbstractMotor protein-driven transport of mRNAs on microtubules and their local translation underlie important neuronal functions such as development, growth cone steering, and synaptic plasticity. While there is abundant data on how membrane-bound cargoes such as vesicles, endosomes, or mitochondria are coupled to motor proteins, surprisingly little is known on the direct interactions of RNA–protein complexes and kinesins or dynein. Provided the potential building blocks are identified, in vitro reconstitutions coupled to Total Internal Reflection Microscopy (TIRF-M) are a powerful and highly sensitive tool to understand how single molecules dynamically interact to assemble into functional complexes. Here we describe how we assemble TIRF-M imaging chambers suitable for the imaging of single protein–RNA complexes. We give advice on optimal sample preparation procedures and explain how a minimal axonal mRNA transport complex can be assembled in vitro. As these assays work at picomolar-range concentrations of proteins and RNAs, they allow the investigation of molecules that cannot be obtained at high concentrations, such as many large or disordered proteins. This now opens the possibility to study how RNA-binding proteins (RBPs), RNAs, and microtubule-associated proteins act together in real-time at single-molecule sensitivity to create cytoplasmic mRNA distributions.
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Breitsprecher, Dennis, Antje K. Kiesewetter, Joern Linkner, and Jan Faix. "Analysis of Actin Assembly by In Vitro TIRF Microscopy." In Methods in Molecular Biology, 401–15. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60761-198-1_27.

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Stoppin-Mellet, Virginie, Nassiba Bagdadi, Yasmina Saoudi, and Isabelle Arnal. "Studying Tau-Microtubule Interaction Using Single-Molecule TIRF Microscopy." In Methods in Molecular Biology, 77–91. New York, NY: Springer US, 2019. http://dx.doi.org/10.1007/978-1-0716-0219-5_6.

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Ho, Chih-Hu, Vladimir Hlady, Chen-Ze Hu, and E. Kurt Dolence. "Assaying Primary Amines on Modified Polymer Surfaces Using TIRF Spectroscopy." In Surface Modification of Polymeric Biomaterials, 97–105. Boston, MA: Springer US, 1996. http://dx.doi.org/10.1007/978-1-4899-1953-3_12.

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Nagamatsu, Shinya, and Mica Ohara-Imaizumi. "Imaging Exocytosis of Single Insulin Secretory Granules With TIRF Microscopy." In Methods in Molecular Biology, 259–68. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-178-9_20.

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Jang, Erika, Siavash Ghaffari, and Warren L. Lee. "Quantifying Endothelial Transcytosis with Total Internal Reflection Fluorescence Microscopy (TIRF)." In Methods in Molecular Biology, 115–24. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2051-9_7.

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Schmidt-Marcec, Sharol, Austin Ross, and Andrei Smertenko. "Quantification of Microtubule-Bundling Activity of MAPs Using TIRF Microscopy." In The Plant Cytoskeleton, 1–12. New York, NY: Springer US, 2023. http://dx.doi.org/10.1007/978-1-0716-2867-6_1.

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Xu, Xuehua, Peter Johnson, and Susette C. Mueller. "Breast Cancer Cell Movement: Imaging Invadopodia by TIRF and IRM Microscopy." In Methods in Molecular Biology, 209–25. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60761-198-1_14.

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Conference papers on the topic "TIRF"

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Huang, Xiaoshuai, Yongxiao li, Weijian Zong, Xiaolu Zheng, Yi Wu, Shiqun Zhao, and liangyi Chen. "MEMS-Based Shadowless-illuminated variable-angle TIRF (SIVA-TIRF)." In CLEO: Applications and Technology. Washington, D.C.: OSA, 2015. http://dx.doi.org/10.1364/cleo_at.2015.ath1j.3.

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Ruckstuhl, Thomas, Michael Rankl, and Stefan Seeger. "Confocal TIRF microscopy of single molecules." In Biomedical Optics 2003, edited by Dan V. Nicolau, Joerg Enderlein, Robert C. Leif, and Daniel L. Farkas. SPIE, 2003. http://dx.doi.org/10.1117/12.485713.

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Saxe, Steven G., and David L. Wortman. "Reflective Performance Of Total Internal Reflection Film (TIRF)." In 31st Annual Technical Symposium, edited by Carl M. Lampert. SPIE, 1987. http://dx.doi.org/10.1117/12.941904.

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Soubies, Emmanuel, Sebastien Schaub, Agata Radwanska, Ellen Van Obberghen-Schilling, Laure Blanc-Feraud, and Gilles Aubert. "A framework for multi-angle TIRF microscope calibration." In 2016 IEEE 13th International Symposium on Biomedical Imaging (ISBI 2016). IEEE, 2016. http://dx.doi.org/10.1109/isbi.2016.7493355.

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Mouttou, Anita, Yousra Toumi, Fabien Lemarchand, Cihan Koc, Antonin Moreau, Delphine Muriaux, Guillaume Demésy, Julien Lumeau, Cyril Favard, and Aude L. Lereu. "Improved TIRF imaging through resonant dielectric multilayer optimization." In Active Photonic Platforms (APP) 2023, edited by Ganapathi S. Subramania and Stavroula Foteinopoulou. SPIE, 2023. http://dx.doi.org/10.1117/12.2676378.

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Blandin, P., S. Lévêque-Fort, S. Lécart, P. Zeller, Z. Lenkei, F. Druon, and P. Georges. "Development of a TIRF-FLIM microscope for biomedical applications." In European Conference on Biomedical Optics. Washington, D.C.: OSA, 2007. http://dx.doi.org/10.1364/ecbo.2007.6630_10.

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Kner, P., B. Chhun, E. Griffis, L. Winoto, L. Shao, and M. G. L. Gustafsson. "Live TIRF microscopy at 100nm resolution through structured illumination." In SPIE BiOS: Biomedical Optics, edited by Jose-Angel Conchello, Carol J. Cogswell, and Tony Wilson. SPIE, 2009. http://dx.doi.org/10.1117/12.812351.

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Blandin, P., S. Lévêque-Fort, S. Lécart, P. Zeller, Z. Lenkei, F. Druon, and P. Georges. "Development of a TIRF-FLIM microscope for biomedical applications." In European Conference on Biomedical Optics, edited by Tony Wilson and Ammasi Periasamy. SPIE, 2007. http://dx.doi.org/10.1117/12.728429.

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Kim, J. W., and K. Kim. "Response-Based Evaluation of Design Sloshing Loads for Membrane-Type LNG Carriers." In ASME 2007 26th International Conference on Offshore Mechanics and Arctic Engineering. ASMEDC, 2007. http://dx.doi.org/10.1115/omae2007-29746.

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Sloshing model test and dynamic structural analysis of a membrane-type LNG tank is performed to determine design sloshing loads for LNG containment system. Spatial, temporal and statistical characteristics of the measured sloshing loads are investigated. Dynamic structural analysis of a LNG containment system is performed to obtain structural responses at predefined critical locations under short duration triangular pulse, which is referred to as triangular impulse response function (TIRF). The TIRFs are synthesized with time history of measured sloshing loads to obtain dynamic response of the LNG containment system. Statistical analysis of peak stress values are used as basis of determining design sloshing loads for the structural assessment of LNG containment system.
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Wu, Xiangping, Xiaofang Liu, Wenglong Xu, Dandan Yan, and Yongli Chen. "Particle filtering for tracking of GLUT4 vesicles in TIRF microscpy." In Sixth International Symposium on Multispectral Image Processing and Pattern Recognition, edited by Jianguo Liu, Kunio Doi, Aaron Fenster, and S. C. Chan. SPIE, 2009. http://dx.doi.org/10.1117/12.831433.

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Reports on the topic "TIRF"

1

Lula, J. W., and G. W. Bohnert. Scrap tire recycling. Office of Scientific and Technical Information (OSTI), March 1997. http://dx.doi.org/10.2172/491404.

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Parsons, G., J. Rafferty, and S. Zilles. Tag Image File Format (TIFF) - image/tiff MIME Sub-type Registration. RFC Editor, March 1998. http://dx.doi.org/10.17487/rfc2302.

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Parsons, G., and J. Rafferty. Tag Image File Format (TIFF) - image/tiff MIME Sub-type Registration. RFC Editor, September 2002. http://dx.doi.org/10.17487/rfc3302.

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NAVAL POSTGRADUATE SCHOOL MONTEREY CA. Land Vehicle Tire Qualification. Fort Belvoir, VA: Defense Technical Information Center, February 2008. http://dx.doi.org/10.21236/ada489076.

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Vinkovich, Richard. Land Vehicle Tire Qualification. Fort Belvoir, VA: Defense Technical Information Center, March 2009. http://dx.doi.org/10.21236/ada496846.

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Al-Qadi, Imad, Jaime Hernandez, Angeli Jayme, Mojtaba Ziyadi, Erman Gungor, Seunggu Kang, John Harvey, et al. The Impact of Wide-Base Tires on Pavement—A National Study. Illinois Center for Transportation, October 2021. http://dx.doi.org/10.36501/0197-9191/21-035.

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Researchers have been studying wide-base tires for over two decades, but no evidence has been provided regarding the net benefit of this tire technology. In this study, a comprehensive approach is used to compare new-generation wide-base tires (NG-WBT) with the dual-tire assembly (DTA). Numerical modeling, prediction methods, experimental measurements, and environmental impact assessment were combined to provide recommendations about the use of NG-WBT. A finite element approach, considering variables usually omitted in the conventional analysis of flexible pavement was utilized for modeling. Five hundred seventy-six cases combining layer thickness, material properties, tire load, tire inflation pressure, and pavement type (thick and thin) were analyzed to obtained critical pavement responses. A prediction tool, known as ICT-Wide, was developed based on artificial neural networks to obtain critical pavement responses in cases outside the finite element analysis matrix. The environmental impacts were determined using life cycle assessment. Based on the bottom-up fatigue cracking, permanent deformation, and international roughness index, the life cycle energy consumption, cost, and green-house gas (GHG) emissions were estimated. To make the outcome of this research effort useful for state departments of transportation and practitioners, a modification to AASHTOWare is proposed to account for NG-WBT. The revision is based on two adjustment factors, one accounting for the discrepancy between the AASHTOware approach and the finite element model of this study, and the other addressing the impact of NG-WBT.
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Bauman, B. D. High Value Scrap Tire Recycle. Office of Scientific and Technical Information (OSTI), February 2003. http://dx.doi.org/10.2172/895571.

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Zamecnik, Robert. Tire Pyrolysis Feasibility Study Approach. Office of Scientific and Technical Information (OSTI), March 2016. http://dx.doi.org/10.2172/1482998.

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Tielking, John T. Aircraft Tire/Pavement Pressure Distribution. Fort Belvoir, VA: Defense Technical Information Center, June 1989. http://dx.doi.org/10.21236/ada279100.

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McIntyre, L., G. Parsons, and J. Rafferty. Tag Image File Format Fax eXtended (TIFF-FX) - image/tiff-fx MIME Sub-type Registration. RFC Editor, September 2002. http://dx.doi.org/10.17487/rfc3250.

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