Off-campus South Dakota State University users: To download campus access theses, please use the following link to log into our proxy server with your South Dakota State University ID and password.

Non-South Dakota State University users: Please talk to your librarian about requesting this thesis through interlibrary loan.

Document Type

Dissertation - University Access Only

Award Date

2015

Degree Name

Doctor of Philosophy (PhD)

Department

Chemistry and Biochemistry

First Advisor

Adam D. Hoppe

Abstract

Fluorescence microscopy Protein-protein interactions Cells

Library of Congress Subject Headings

Quantitative Fluorescence Resonance Energy Transfer (FRET) microscopy is capable of imaging the nanoscale associations of proteins in living cells. These methods have opened new windows to visualize and quantify when and where proteins are touching and conducting biochemistry within the cell, thereby providing a powerful tool to translate the current understanding of in vitro biochemistry into its native context. Until recently, FRET microscopy approaches have been limited to the analysis of one protein-protein interaction at a time and in two-dimensions. In this dissertation, FRET microscopy methods are developed which are capable of imaging the interactions of any number of fluorophores in two-dimensions. This dissertation also presents the first image reconstruction approach to improve the three-dimensional optical sectioning of multiple protein-protein interaction. Additionally, an in-depth characterization of fluorescent proteins for multifluorophore FRET was conducted leading to the selection of an optimized fluorescent protein trio that enhances the sensitivity and reliability of the new approaches. Together this work represents a significant advance in methods for studying biochemistry within the context of the living cell.

Description

Includes bibliographical references

Format

application/pdf

Number of Pages

154

Publisher

South Dakota State University

Rights

In Copyright - Educational Use Permitted
http://rightsstatements.org/vocab/InC-EDU/1.0/

Share

COinS