Single-molecule imaging studies of protein dynamics

Single-molecule fluorescence imaging is a powerful method for studying biological events. The work of this thesis primarily focuses on single molecule studies of the dynamics of Green Fluorescent Protein: GFP) and other fluorescent-labeled proteins by utilizing Total Internal Reflection Fluorescence: TIRF) microscopy and imaging. The single molecule experiments of this thesis covered three broad topics. First, the adsorption mechanisms of proteins onto hydrophobic and hydrophilic fused silica surfaces were imaged and reversible and irreversible adsorption mechanisms were observed. The second topic covered a new technique for measuring the diffusion coefficient of Brownian diffusing proteins, in particular GFP, in solution via a single image. The corresponding experiments showed a relationship between the intensity profile width and the diffusion coefficient of the diffusing molecules. The third topic covered an in vivo experiment involving imaging and quantifying prokaryotic cell metabolism protein dynamics inside the Bacillus subtilis bacteria, in which a helical diffusion pattern for the protein was observed. These topics are presented in the chronological order of the experiments conducted.

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Physics

Year dc:date.available

2011

Author dc:creator

• Zareh, Shannon Kian

Contributors dc:contributor

• YAN MEI WANG

• Physics
• Biophysics
• DIFFUSION COEFFICEINT
• FLUORESCENCE IMAGING
• GAUSSIAN DISTRIBUTION STANDARD DEVIATION
• SINGLE MOLECULE
• TIRF MICROSCOPY

Language dc:language

English (en)

DOI

https://doi.org/10.7936/K7X06556

OAI identifier oai:identifier

oai:openscholarship.wustl.edu:etd-1672