Global ETD Search
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Showing 1 to 14 of 14 for “"SMLM"”.
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Applying single-molecule localisation microscopy to achieve virtual optical sectioning and study T-cell activation
Single-molecule localisation microscopy (SMLM) allows imaging of fluorescently-tagged proteins in live cells with a precision well below that of the diffraction limit. As a single-molecule technique, it has also introduced a new quantitative approach to fluorescence microscopy. In the Part A of …
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Advances in volumetric super-resolution microscopy and single-particle tracking
Single-molecule localisation microscopy (SMLM) has enabled optical microscopy to probe biological structures and processes at the nanometer scale. To accommodate the inherently 3D nature of biological structures, 3D-SMLM techniques---such as the astigmatic and double-helix point spread functions …
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Single Molecule Localisation with Fourier Light Field Microscopy
… of 3D Single Molecule Localisation Microscopy (SMLM). This technique combines the super-resolution capability of SMLM with the principles of parallax; multiple perspective views of photo-switchable fluorophores are captured using a lenslet array in the Fourier plane of a fluorescence microscope. …
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Super-Resolved Microscopy as a Tool to Unveil Intracellular Structure in Biological Systems
… These single-molecule localization microscopy (SMLM) methods can achieve a resolution of roughly 20 nm. In addition, we developed image-processing tools to reconstruct accurate SMLM images and studied the photo-physics of various photo-switchable fluorophores to confirm their suitability for …
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Development of Three-Dimensional Super-Resolution Imaging Using a Double-Helix Point Spread Function
Single-molecule localisation microscopy (SMLM), has allowed for optical microscopy to probe biological systems beyond the diffraction limit. The intrinsic 3D nature of biology has motivated the development of 3D-SMLM with novel techniques, including the double-helix point spread function (DHPSF). A …
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Vortex Light Field Microscopy for 3D Spectral Single-Molecule Imaging
Singe-molecule localization microscopy (SMLM) is a powerful imaging technique that surpasses the diffraction limit and significantly improves spatial resolution over conventional light microscopy, allowing cellular structures and dynamic processes to be visualized at nanoscale. To solve the …
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Convex Relaxations for Particle-Gradient Flow with Applications in Super-Resolution Single-Molecule Localization Microscopy
<p>Single-molecule localization microscopy (SMLM) techniques have become advanced bioanalytical tools by quantifying the positions and orientations of molecules in space and time at the nanoscale. With the noisy and heterogeneous nature of SMLM datasets in mind, we discuss leveraging …
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Advancing Fluorescence Microscopy Techniques for Volumetric Whole-cell Imaging
… robust single-molecule localisation microscopy (SMLM) pipeline for volumetric imaging in the whole cell using the double-helix point spread function (DH-PSF) approach. The practical challenges associated with this method, such as aberration- and drift-induced inaccuracies, were comprehensively …
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Synthetic ground truth of biological shapes — Simulating variable Nuclear Pore Complexes for Microscopy
… Single Molecule Localization Microscopy (SMLM) images NPCs at protein-level resolution, whereupon image analysis methods study NPC variability. However, the true picture of NPC variability is unknown and the biological function of this variability is poorly understood. In quantitative …
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Development of Novel Labelling Strategies for Imaging in Nanoscale Topography
… 3D single molecule localisation microscopy (3D-SMLM) techniques such as the double helix point spread function (DHPSF) enable the visualisation of these systems on the required spatial scales and take into account the inherently 3D nature of biology. However, the addition of axial information …
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Three-Dimensional Single-Molecule Localisation Microscopy and Visualisation Techniques
… for single-molecule localisation microscopy (SMLM), we were able to explore, segment, analyse and export data through an intuitive medium. This thesis presents the first DH-PSF brain tissue imaging to understand the organisation of a scaffolding protein known as postsynaptic density 95 …
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Optical imaging methods for the study of disease models from the nano to the mesoscale
… limit. Single-molecule localisation microscopy (SMLM), which circumvents the diffraction limit by separating fluorophore emission in time to localise individual molecules in space with ∼20 nm precision, was thus implemented to study a-syn in purified synaptic boutons. A software package was …
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Single-molecule orientation estimation using a polarisation camera
This thesis presents the development of a new optical imaging method for measuring the orientation and position of single fluorescent molecules. The method - named POLCAM - is based on the use of a polarisation camera. Current methods for single-molecule orientation estimation require optical …