Multi Scale Fluorescence Studies: The Spinning Disk Confocal Fluorescence Lifetime Imaging Microscopy &Amp; Single Molecule Nano-Metronome &Amp; Single Molecule Study of Tetramethyrhodamine-Dna Interaction

The thesis is consisted of three different sections independent of each other. The first part involves the frequency domain fluorescence lifetime imaging microscopy (FLIM) and an improvement of FLIM setup to decrease the out of focus background fluorescence without drastically sacrificing the data acquisition speed while taking fluorescence lifetime data. The setup incorporates a spinning disk confocal unit to the conventional wide-field fluorescence lifetime imaging; therefore, it is a marriage between confocality and data acquisition speed. The detailed discussion about the principle of the setup in conjunction to the benefits when the setup is used for imaging thick specimens is presented. In addition, a new concept of using the Wavelets analysis on fluorescence lifetime images is introduced. Besides facilitating morphology recognition and noise reduction usually shown in normal image analysis, the benefit of wavelets analysis on decomposing an image into different spatial frequencies proves to be useful in removing the lifetime information of the background region out of the lifetime information from the small structures of interest. Moreover, the solutions to the artifacts often encountered in frequency domain fluorescence lifetime measurements are shown. Finally, the FLIM setup is used to study the close interactions between structural proteins in the body wall muscle cells of the nematode Caenorhabditis elegans and visualize the activation states of a signaling protein in cultured neurons of Drosophila melanogaster based on the change in the fluorescence lifetime of CFP at different energy transfer efficiencies between the CFP and YFP pair.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biophysics and Computational Biology

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Buranachai, Chittanon

Contributors dc:contributor

• Clegg, Robert M.

• Biophysics, General

Identifier

(UMI)AAI3314736

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/72394