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Title

Advances in Fluorescence Correlation Spectroscopy with Applications to Studies of Macromolecular Interactions and Folding of RNA Hairpins

Date of Award

Summer 8-15-2010

Author's School

Graduate School of Arts and Sciences

Author's Department

Biology & Biomedical Sciences (Computational & Molecular Biophysics)

Degree Name

Doctor of Philosophy (PhD)

Degree Type

Dissertation

Abstract

This work is dedicated to developing theory and applications of fluctuation spectroscopy, an experimental technique that allows one to measure and analyze signal fluctuations that are due to single molecules. By averaging many fluctuations, it is possible to obtain information about distributions of molecular properties of the ensemble without disturbing the equilibrium. Our work continues development of widely popular fluorescence correlation spectroscopy. Towards this goal, we develop full theory of two-point correlations in a light detection experiment and revive application of high-order fluorescence correlation spectroscopy in biochemistry. Our theoretical findings lead to simplified data analysis that efficiently captures all important molecular properties: brightness per molecule, diffusion coefficient of the molecules, and relaxation rates of chemical reactions. We are also able to obtain theoretical expressions that describe the signal-to-noise ratio in high-order correlation experiments. The developed methodology is especially useful in investigating irreversible chemical reactions as we show by quantitatively describing photobleaching in the excitation volume.

Language

English (en)

Chair and Committee

Kathleen B. Hall

Committee Members

Eliot Elson, Carl Frieden, Eric A. Galburt, Roberto Galletto, Guy M. Genin

Comments

Permanent URL: https://doi.org/10.7936/K73N21BC

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