Advances in Fluorescence Correlation Spectroscopy with Applications to Studies of Macromolecular Interactions and Folding of RNA Hairpins
Date of Award
Doctor of Philosophy (PhD)
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.
Chair and Committee
Kathleen B. Hall
Eliot Elson, Carl Frieden, Eric A. Galburt, Roberto Galletto, Guy M. Genin
Melnykov, Artem Victorovich, "Advances in Fluorescence Correlation Spectroscopy with Applications to Studies of Macromolecular Interactions and Folding of RNA Hairpins" (2010). Arts & Sciences Electronic Theses and Dissertations. 36.
Permanent URL: https://doi.org/10.7936/K73N21BC