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 18 of 18 for “"total internal reflection fluorescence microscopy"”.
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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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The photophysics properties of lanthanide luminescent probes
… have been carried out using a time- resolved fluorescence assay (TRFA). Bi-cs124 was conbined with different chelating agents, specifically diethylene triamine pentaacetic acid (DTPA), ethylene diamine tetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), and 3,6,9,12-Tetrakis …
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S-Acylation Is A Key Regulator of Orai1/Stim1-Mediated Store-Operated Calcium Entry In T Cells
… and mediate SOCE. Lastly, our studies using total internal reflection fluorescence microscopy showed that a lack of S-acylation of either Orai1 or STIM1 resulted in significantly reduced Orai1/STIM1 colocalization as evident from the diminished PM puncta formation. These data describe a novel …
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Protein segregation and conformation in antigen-receptor triggering: a quantitative fluorescence microscopy study
… distance affects antigen-receptor triggering. Total internal reflection fluorescence microscopy is used to establish that phosphatase exclusion decreases when intermembrane distance increases. This inverse relationship, demonstrated for both B cells and T cells, is consistent with the …
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Biochemical and Microscopic Characterization of INFT-1: an Inverted Formin in C. elegans
… INFT-1 is regulated by other mechanisms. We used fluorescence spectroscopy to determine the effect of INFT-1 FH1FH2 on actin assembly and total internal reflection fluorescence microscopy to investigate how INFT-1 formin homology 1 and formin homology 2 domains (FH1FH2) mediate filament nucleation …
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Mechanistic Analysis of Microtubule Dynamics and Regulation
… all-yeast in vitro reconstitution system using total internal reflection fluorescence microscopy which enabled me to study a variety of Stu2 mutants, in the presence of wild-type and mutant yeast tubulin samples. Here, I discovered how the tubulin conformational state can impact Stu2 function …
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A Platform for Analyzing Force Sensitivity and Multivalency in Actin Networks
… with varying architectures simultaneously using total internal reflection fluorescence microscopy, facilitating detailed dissection of the interplay between force-modulated ABP binding and network geometry. Secondly, I present data applying our system to study a dimeric form of the critical …
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Investigation of physical principles of protein phase transitions
… and elongation events inside condensates using total internal reflection fluorescence microscopy and spinning disk super resolution by optical pixel reassignment microscope. This reveals that the time-dependent changes in the physical and biochemical properties of biomolecular condensates are …
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Actin polymerization dynamics at the leading edge
… elongation and attachment to leading edge. Using total internal reflection fluorescence microscopy of actin filaments, we tested how profilin, Arp2/3, and capping protein (CP) function together to propel beads or thin glass nanofibers coated with N-WASP WCA domains. During sustained motility, …
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Multi-Modal Regualtion of Actin Networks
… simultaneous optical trapping and confocal microscopy assays and in vitro reconstituted myosin motor-based total internal reflection fluorescence microscopy assays were employed to study how mechanical forces applied on actin filaments can regulate actin binding. A case study on a homologous …
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Visualizing the One-Dimensional Diffusion of DNA Mismatch Repair Proteins at the Single-Molecule Level
… of probing these dynamic processes. A custom total internal reflection fluorescence microscopy assay was developed to investigate these events by visualizing the proteins as they search along DNA in real time at the single molecule level. Both Msh2-Msh6 and Mlh1-Pms1 were found to travel along …
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Ultrasensitive quantification of circulating disease biomarkers through enzymatic labeling and single molecule counting
… molecules are then directly counted using total internal reflection fluorescence microscopy, as opposed to conventional indirect methods using the polymerase chain reaction (PCR). This technology provides a fundamental framework for the development of an ultrasensitive miRNA detection …
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Single molecule pull-down of viral RNA-protein complexes
… RNA molecules for detection with single molecule fluorescence microscopy. We demonstrate specific capture of viral DI RNA molecules using SiMPull, and could quantitatively measure the presence of interacting viral proteins. Therefore, this study provides evidence for the applicability of SiMPull …
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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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Single-molecule studies of bacterial DNA replication and translesion synthesis
Faithful replication of genomic DNA is crucial for the survival of a cell. In order to achieve high-level accuracy in copying its genome, all cells employ replicative DNA polymerases that have intrinsic high fidelity. When an error occurs on the template DNA strand, in the form of lesions caused by …
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Untersuchungen in lebenden Zellen mit Hilfe neuer Methoden der Einzelmolekülfluoreszenzspektroskopie
… (engl.: single molecule fluorescence spectroscopy, Abk.: SMFS), zu denen Methoden, wie Totalreflexions-Fluoreszenz Mikroskopie (engl.: total internal reflection fluorescence microscopy, Abk.: TRIFM) und die konfokale Einzelmolekül¬fluoreszenzmikroskopie gehören. Die Stärke …
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Cytotoxic T lymphocyte biology under the microscope
… biology. Throughout these different projects, microscopy has played an important part. Seeing really is believing, and the versatility of microscopic approaches is hopefully on full display throughout this thesis. High resolution live microscopy of individual CTL:target conjugates is perhaps …