Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 19 of 19 for “"Total-Internal-Reflection Fluorescence (TIRF) microscopy"”.
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Single-molecule imaging studies of protein dynamics
Single-molecule fluorescence imaging is a powerful method for studying biological events. The work of this thesis primarily focuses on single molecule studies of the dynamics of Green Fluorescent Protein: GFP) and other fluorescent-labeled proteins by utilizing Total Internal Reflection …
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Direct Single Molecule Imaging of Enhanced Diffusion for Enzymes and Enzyme-conjugated Dna origami
… using single-particle tracking (SPT) with total internal reflection fluorescence (TIRF) microscopy. We found that the mobility of individual enzymes was enhanced three-fold in the presence of the substrate, and the motion remained Brownian. We showed that the relative increase in diffusion …
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Domain Redistribution in Lipid Bilayers in the Presence of the Antimicrobial Lipopeptide Fengycin
… cyclic lipopeptide fengycin on model membranes. Total internal reflection fluorescence (TIRF) microscopy was used to visualize and analyse lipid phase separation within supported lipid bilayers (SLBs) of various compositions. The initial research investigated the influence of protocol parameters …
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Probing cellular protein complexes using single-molecule pull-down
… biomolecules at single-molecule resolution using total internal reflection fluorescence (TIRF) microscopy. We name this technology single-molecule pull-down or SiMPull. In one reification biotinylated antibody against the protein of interest is immobilized on a flow chamber. The flow chambers are …
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Investigating Spatiotemporal Kinetics, Dynamics, and Mechanism of Exosome Release
… cells (A549) using CD63 fluorescent probes and total internal reflection fluorescence (TIRF) microscopy. The kinetics of release, or loss of fluorescence post-fusion, can relay information about the fate of exosomes. Using kinetic analysis we determined that a portion of exosomes are free to …
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Microfluidic chips for combinatorial screening applications
… of detecting complimentary DNA fragments. Total internal reflection fluorescence (TIRF) microscopy is used for read out of the chip. Four virus-like oligomer targets with sequences corresponding to key fragments of the viruses HIV, HPV, Hepatitis A and Hepatitis B were tested against four …
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Development of tools for single molecule peptide fluorosequencing
… fluorophore labeling of amino acid residues, total-internal reflection fluorescence (TIRF) microscopy, and Edman degradation, is a novel technology that has been developed to sequence peptides. Unique fluorophores are covalently attached to amino acid residues and through TIRF microscopy are …
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Quantifying the Release of Protein Substrates from AAA+ ATPase ClpX by Single Molecule Total Internal Reflection Fluorescence Microscopy
… ATPase ClpXP. We present a new assay based on Total-Internal-Reflection Fluorescence (TIRF) microscopy that measures the lifetime of the ClpX-substrate complex. We demonstrate that the technique has the potential to identify factors that affect the mean lifetime of the ClpX-substrate complex. …
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USING SINGLE VIRION FUSION ASSAY TO STUDY INFLUENZA VIRUS ENTRY
… each of these processes in vitro. SPT combines total internal reflection fluorescence (TIRF) microscopy with microfluidics and supported lipid bilayers, making it a very powerful tool for host-pathogen membrane fusion studies. One concern about influenza is that the virus can mutate quickly via …
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MEMS-based temperature-dependent characterization of biomolecular interactions
… handling, on-chip temperature control, and total internal reflection fluorescence (TIRF) microscopy, we have studied the temperature dependence of the structural dynamics of transfer RNA (tRNA) translocation through ribosome in protein synthesis. For solution-based biomolecular interactions, …
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High-resolution optical analyses of IP3-evoked Ca2+ signals
… of caged- IP3 were recorded using Cal520 and total internal reflection fluorescence (TIRF) microscopy in human embryonic kidney (HEK) cells. The Flika algorithm was used, and validated, for automated detection of Ca2+ puffs and to measure their properties. IP3 evoked Ca2+ puffs in wild-type …
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Investigating the molecular pharmacology of class A and B G protein-coupled receptors
… receptor activity-modifying proteins (RAMPs), or internalisation, whereby distinct responses may be propagated by intracellular GPCR populations. Hence, the signalling repertoire subsequent to receptor activation is extremely complex, varying between individual GPCRs and the biological systems in …
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Development of the single-molecule tracking and fluorescence lifetime techniques for biophysical measurements and biomedical applications
… 3). In Chapter 4, I further employed 3D-SMT and fluorescence correlation spectroscopy (FCS) to characterize the diffusivity of an individual JNK2 oligomers in live HeLa cells and demonstrated that JNK2 tetramers can be stable inside mammalian cells. Although total internal reflection fluorescence …
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Examining Mechanisms Regulating Microtubule Assembly and Function
… composition. I used in vitro reconstitution and total internal reflection fluorescence (TIRF) microscopy to examine the dynamics of individual microtubules assembled from these distinct heterodimers. I found that microtubules assembled with βIIB are substantially more stable, switching from a …
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The Roles of Focal Adhesion Assembly in Membrane Localization of Ion Channels and Action Potential Shape
… with single cell electrophysiology, confocal and total internal reflection fluorescence (TIRF) microscopy, and quantitative image analysis, we provide evidence that FAs and active integrins play a critical role in regulating Kir2.1 membrane localization, IK1 amplitude, and action potential …
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Breaking Down Microtubule Formation: Characterizing the Biochemical and Cellular Functions of the γ-Tubulin Ring Complex
… event, or on pre-formed microtubules. Using total internal reflection fluorescence (TIRF) microscopy, the dynamics of this association can be quantified in terms of the number of microtubules that are capped, and the length of time the cap persists at the minus-end under these two conditions. …
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Investigating TLR-4 signalling in response to protein ligands
… fixed at different time points, then imaged. Total internal reflection fluorescence (TIRF) microscopy was used to image the plasma membrane and photobleaching experiments performed to determine Mal/TIRAP stoichiometry. I developed a computer-based analysis pipeline to analyse the resulting …
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High-resolution optical analyses of inositol 1,4,5-trisphosphate receptors and the Ca²⁺ puffs they evoke
… endogenous IP₃Rs and using high-resolution total internal reflection fluorescence (TIRF) microscopy, I was able to visualise both IP₃Rs and the Ca²⁺ puffs they evoke following photolysis of a caged analogue of IP₃. I optimised fluorescent labelling of SNAP-IP₃R3, and confirmed that its …