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BL
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| Record Number
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865322
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| Main Entry
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Arroyo, Jaime Ortega
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| Title & Author
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Investigation of nanoscopic dynamics and potentials by interferometric scattering microscopy /\ Jaime Ortega Arroyo.
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| Publication Statement
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Cham, Switzerland :: Springer,, 2018.
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| Series Statement
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Springer theses
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| Page. NO
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1 online resource (xxxvi, 142 pages) :: illustrations (some color)
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| ISBN
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3319770950
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: 9783319770956
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3319770942
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9783319770949
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| Notes
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"Doctoral thesis accepted by the University of Oxford, Oxford, UK."
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| Bibliographies/Indexes
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Includes bibliographical references.
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| Contents
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Intro; Supervisor's Foreword; Abstract; Parts of this thesis have been published in the following journal articles:; Acknowledgements; Contents; List of Figures; 1 Introduction; References; 2 Non-fluorescent Single-Molecule Approaches to Optical Microscopy; 2.1 Introduction; 2.2 Single-Particle Tracking; 2.3 Scattering Detection: An Alternative to Fluorescence; 2.4 Interferometric Scattering; 2.4.1 Confocal Detection; 2.4.2 Non-scanned Wide-Field Detection; 2.4.3 Confocal Beam Scanning Wide-Field Detection; 2.5 Applications; 2.5.1 Lateral Single-Particle Tracking
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2.5.2 Axial Localisation via Interferometry2.5.3 Label-Free Imaging; 2.6 Conclusion and Outlook; References; 3 Experimental Methods; 3.1 Experimental Optics and Hardware; 3.1.1 Interferometric Scattering Channel; 3.1.2 Focus Control Feedback Channel; 3.1.3 Single-Molecule Fluorescence Channel; 3.1.4 Sample Stage Stabilisation; 3.1.5 Camera Characterisation; 3.1.6 Operation and Synchronisation of the Acousto-Optic Beam Deflector; 3.1.7 Data Acquisition; 3.2 Experimental iSCAT Microscopy; 3.2.1 Image Processing; 3.2.2 Spot Detection; 3.2.3 Localisation; 3.2.4 Trajectory Linking
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3.2.5 Assessment of Localisation Precision3.2.6 Self-referencing; References; 4 Anomalous Diffusion Due to Interleaflet Coupling and Molecular Pinning; 4.1 Introduction; 4.2 Experimental Methods; 4.2.1 Materials; 4.2.2 Vesicle Preparation; 4.2.3 Substrate Preparation; 4.2.4 Supported Lipid Bilayer Formation; 4.2.5 Instrument Setup Parameters; 4.3 Experimental Results; 4.3.1 GM1 Undergoes Anomalous Diffusion in Supported Lipid Bilayers; 4.3.2 Transient Confinement Causes Anomalous Diffusion; 4.3.3 Concentration Dependent Dynamics of Transient Binding
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4.3.4 Recovery of Brownian Motion upon Tuning Substrate Interactions and Interleaflet Coupling4.4 Discussion; 4.4.1 Importance of Simultaneous Localisation Precision and Time Resolution; 4.4.2 Thermal and Optical Force Considerations; 4.4.3 Membrane Defects, Labelling Artefacts, and CTxB Induced Aggregation Do Not Cause Transient Binding; 4.4.4 Transient Binding Requires Substrate Interaction and Interleaflet Coupling; 4.4.5 Multiple CTxB-GM1 Interactions Result in Ring-Like Structures; 4.4.6 A Model of Transient Binding: Molecular Pinning; 4.5 Conclusion and Outlook; References
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5 Structural Dynamics of Myosin 5a5.1 Introduction; 5.2 Experimental Methods; 5.2.1 Sample Preparation; 5.2.2 Experimental Setup; 5.3 Experimental Results; 5.3.1 N-Terminus Labelling Does Not Perturb the Kinetics of Myosin 5a; 5.3.2 During Myosin Movement the Motor Domain Undergoes a Transition Between Two Distinct States; 5.3.3 The Labelled Motor Domain Moves in Three Dimensions; 5.3.4 A Conformational Change in the Motor Domain Accompanies the Power Stroke; 5.3.5 Myosin Steps via a Single, Spatially-Constrained Transient State
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| Abstract
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This thesis offers a unique guide to the development and application of ultrasensitive optical microscopy based on light scattering. Divided into eight chapters, it covers an impressive range of scientific fields, from basic optical physics to molecular biology and synthetic organic chemistry. Especially the detailed information provided on how to design, build and implement an interferometric scattering microscope, as well as the descriptions of all instrumentation, hardware interfacing and image processing necessary to achieve the highest levels of performance, will be of interest to researchers now entering the field.
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| Subject
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Interferometry.
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| Subject
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Microscopy.
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Atomic molecular physics.
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Cellular biology (cytology)
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Interferometry.
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Microscopy.
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| Subject
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NATURE-- Reference.
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Organic chemistry.
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Physical chemistry.
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| Subject
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SCIENCE-- Life Sciences-- Biology.
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| Subject
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SCIENCE-- Life Sciences-- General.
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| Dewey Classification
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570.28/2
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| LC Classification
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QH205.2
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