The objective of the proposed research is to develop a novel class of fluorescent nanoparticles called """"""""Polymer Dots"""""""" and test the feasibility of the nanoparticles for detection of single molecules in live cells using conventional fluorescence microscopy methods. Polymer dot nanoparticles are a promising new fluorescent nanoparticle technology based on fluorescent pi-conjugated polymers which provide substantial improvements in brightness as compared to conventional fluorescent dyes and nanoparticles.
The specific aims of the project include optimizing and characterizing relevant figures of merit for single molecule detection, development and testing of bioconjugation strategies for targeting specific biomolecules within cells, and demonstration of single nanoparticle detection in living cells. A range of characterization techniques will be employed, including fluorescence spectroscopy, single molecule spectroscopy, atomic force microscopy, electrophoresis, and epifluorescence microscopy. Improvements in brightness and photostability of a factor of 100 to 10,000 as compared to conventional fluorescent dyes are expected, meeting the requirements for fluorescence-based single molecule detection in living cells. Demonstration of facile, flexible bioconjugation methods for targeting specific biomolecules of interest and detection of individual labeled biomolecules within living cells are also expected. The development of this novel fluorescent nanoparticle platform will facilitate future research involving monitoring transport and biochemical events of single molecules within living cells and could also provide the basis for the development of novel bioassays and biosensors.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM081040-03
Application #
7684801
Study Section
Special Emphasis Panel (ZRG1-BST-R (51))
Program Officer
Deatherage, James F
Project Start
2007-09-01
Project End
2011-08-31
Budget Start
2009-09-01
Budget End
2010-08-31
Support Year
3
Fiscal Year
2009
Total Cost
$248,280
Indirect Cost
Name
Clemson University
Department
Chemistry
Type
Schools of Engineering
DUNS #
042629816
City
Clemson
State
SC
Country
United States
Zip Code
29634
Dukes, Kyle D; Christensen, Kenneth A; Chumanov, George (2014) Core-shell silver nanoparticles for optical labeling of cells. Anal Biochem 458:43-8
Fernando, Lawrence P; Kandel, Prakash K; Ackroyd, P Christine et al. (2012) The relative brightness of PEG lipid-conjugated polymer nanoparticles as fluid-phase markers in live cells. Anal Bioanal Chem 404:3003-14
Yu, Jiangbo; Wu, Changfeng; Tian, Zhiyuan et al. (2012) Tracking of single charge carriers in a conjugated polymer nanoparticle. Nano Lett 12:1300-6
Wu, Changfeng; Hansen, Stacey J; Hou, Qiong et al. (2011) Design of highly emissive polymer dot bioconjugates for in vivo tumor targeting. Angew Chem Int Ed Engl 50:3430-4
Kandel, Prakash K; Fernando, Lawrence P; Ackroyd, P Christine et al. (2011) Incorporating functionalized polyethylene glycol lipids into reprecipitated conjugated polymer nanoparticles for bioconjugation and targeted labeling of cells. Nanoscale 3:1037-45
Fernando, Lawrence P; Kandel, Prakash K; Yu, Jiangbo et al. (2010) Mechanism of cellular uptake of highly fluorescent conjugated polymer nanoparticles. Biomacromolecules 11:2675-82
Wu, Changfeng; Schneider, Thomas; Zeigler, Maxwell et al. (2010) Bioconjugation of ultrabright semiconducting polymer dots for specific cellular targeting. J Am Chem Soc 132:15410-7
Yu, Jiangbo; Wu, Changfeng; Sahu, Sushant P et al. (2009) Nanoscale 3D tracking with conjugated polymer nanoparticles. J Am Chem Soc 131:18410-4
Wu, Changfeng; Bull, Barbara; Christensen, Kenneth et al. (2009) Ratiometric single-nanoparticle oxygen sensors for biological imaging. Angew Chem Int Ed Engl 48:2741-5
Wu, Changfeng; Bull, Barbara; Szymanski, Craig et al. (2008) Multicolor conjugated polymer dots for biological fluorescence imaging. ACS Nano 2:2415-23

Showing the most recent 10 out of 13 publications