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 20 of 31 for “"TIRF MICROSCOPY"”.
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Development of selective high-affinity fluorescent sensors for imaging neurotransmitters and application in cells and tissue
… of total internal reflective fluorescence (TIRF) microscopy and amperometry. A NIR fluorescent sensor NS659 was developed based on the structure of NS510. Several different synthetic routes were attempted before the sensor was successfully prepared. Upon binding to serotonin, the sensor gave …
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Development of tools for single molecule peptide fluorosequencing
… 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 traced through Edman degradation cycles …
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Anomalous diffusion of mRNA in the cytoplasm of HeLa cells
… in live cells via total internal reflection (TIRF) microscopy. In order to visualize individual RNA molecules expressing the MYH9 gene, they were labeled via MS2 stem loops bound to coat proteins tagged with the HaloTag-JF646 fluorophore. We then used single-particle tracking to obtain …
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Single-molecule imaging studies of protein dynamics
… Total Internal Reflection Fluorescence: TIRF) microscopy and imaging. The single molecule experiments of this thesis covered three broad topics. First, the adsorption mechanisms of proteins onto hydrophobic and hydrophilic fused silica surfaces were imaged and reversible and irreversible …
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Serine 910 Phosphorylation of Focal Adhesion Kinase Is Critical for Costamere Assembly
… pathways. Using co-immunoprecipitation, TIRF-microscopy and FRAP, ET-1 stimulation of NRVM expressing a nonphosphorylatable, S910A-FAK mutant decreased the interaction of paxillin and vinculin within costameres. This interaction was important in stabilizing α-actinin within the protein …
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Direct Single Molecule Imaging of Enhanced Diffusion for Enzymes and Enzyme-conjugated Dna origami
… 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 is independent of the total enzyme …
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Domain Redistribution in Lipid Bilayers in the Presence of the Antimicrobial Lipopeptide Fengycin
… 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 on fluorescent probe distribution and …
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Probing cellular protein complexes using single-molecule pull-down
… 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 passivated using a polymer coating such …
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Development of a Model System for Culturing Neonatal Rat Ventricular Myocytes in vitro to Monitor Integrin Activation in Response to Altered Contractile Force
… integrins at cell-ECM contacts using confocal microscopy. Utilizing total internal fluorescence (TIRF) microscopy we have completed preliminary measurements of intensity changes overtime at focal adhesions. Utilizing ImageJ technology, we aim to determine how changes in acto-myosin force affect …
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Biophysical characterization of the Escherichia coli RNA degradosome
… study. Total internal reflection fluorescent (TIRF) microscopy of the RNA degradosome fluorescently labeled using a HaloTag showed discrete particles of the enzyme complex on a supported lipid bilayer on glass. To enable in situ studies of the degradosome, the HaloTag was encoded at the end of …
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Investigating Spatiotemporal Kinetics, Dynamics, and Mechanism of Exosome Release
… 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 diffuse away from the fusion site, but some …
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Microfluidic chips for combinatorial screening applications
… 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 different molecular beacons, each …
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Single molecule fluorescence studies of biomolecular interactions
… this question we combined single molecule TIRF microscopy and multi-color fluorescent in situ hybridization (FISH) to study the composition of viral RNAs at single-virus particle resolution. Our results showed that a high percentage of virus particles package a single copy of each segment …
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Quantifying the Release of Protein Substrates from AAA+ ATPase ClpX by Single Molecule Total Internal Reflection Fluorescence Microscopy
… 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. Using this method, we show that the …
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USING SINGLE VIRION FUSION ASSAY TO STUDY INFLUENZA VIRUS ENTRY
… 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 three mechanisms (i.e. error-prone RNA …
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MEMS-based temperature-dependent characterization of biomolecular interactions
… 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, we mainly focus on calorimetry, a …
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High-resolution optical analyses of IP3-evoked Ca2+ signals
… 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 HEK cells and in cells expressing single …
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Conformational Dynamics of the Nuclear Pore Complex Studied With Coarse-Grained Modelling and Polarized Light Microscopy
… internal reflection fluorescence (pol-TIRF) microscopy. The NPC is a large (~120 MDa in humans) macromolecular assembly that controls the flow of molecules into and out of the nucleus. The NPC is a key regulator of intracellular trafficking, nuclear organization, and gene expression. …
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Single molecule investigation of antiviral signaling
… process for IFN- production. Fluorescence microscopy is an essential tool in biology. However, fluorescence microscopy is limited by diffraction to a spatial resolution of >200 in lateral dimensions, and >500 nm in axial dimension, leaving many biological structures too small to study in …
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Investigating the molecular pharmacology of class A and B G protein-coupled receptors
… bioluminescent resonance energy transfer (BRET) microscopy. Subsequently, the effect(s) of RAMP1 mutations upon agonist-mediated signalling is investigated in terms of the abilities of mutant RAMP1-CLR heterodimers to mediate cAMP production and β-arrestin recruitment, in various cell types. Such …
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