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Document Type
Dissertation - University Access Only
Award Date
2015
Degree Name
Doctor of Philosophy (PhD)
Department / School
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
Recommended Citation
Scott, Brandon L., "Advances in FRET Microscopy for the Analysis of multiple Protein-protein Interactions in the Three-dimensional Space of the Living Cell" (2015). Electronic Theses and Dissertations. 1878.
https://openprairie.sdstate.edu/etd/1878
Rights Statement
