Fluorescence based techniques, from cellular imaging to bioassays, have become an indispensable tool kit in both basic cellular studies of cancer and in clinical applications and in vitro diagnostics. Organic dyes appear to be the most versatile fluorophores used so far in these applications. However, the intrinsic limitations of conventional dyes, such as low absorptivity and poor photostability, have posed difficulties for the further development of high-sensitivity imaging techniques and high-throughput assays. As a result, there has been considerable interest in brighter and more stable fluorescent probes. For example, phycobiliproteins exhibit higher fluorescence brightness than small organic fluorophores, and there is also a great deal of interest in the development of brightly fluorescent nanoparticles, such as semiconductor quantum dots. This project aims to develop a new class of fluorescent probes, called Pdots, for use in the molecular analysis and study of cancer cells. To achieve this goal, we propose the following three aims:
Aim 1 : Develop multicolor ultra-bright Pdots with ultra-narrow emission bandwidth (Full Width at Half Maximum (FWHM) < 15 nm). Although Pdots are exceptionally bright, a severe drawback is the very broad emission spectra of currently available Pdot species, which limit their usefulness in multiplexed applications. There is an urgent need to develop new types of Pdots which can emit at different wavelengths with a narrow-band spectral width. Here, we propose to develop a series of Pdots with an ultra-narrow emission bandwidth of <15nm.
Aim 2 : Generation of Pdots with monodisperse size distributions. Although we can tune the size of Pdots from about 5nm to tens of nanometers in diameter, their size distribution is currently quite broad. Here, we proposed to develop an efficient and high-throughput method based on monodisperse filter pores to form Pdots of monodisperse size distributions.
Aim 3 : Generation of monovalent single-chain Pdots with controlled surface properties. It is often extremely difficult to control the number and geometrica distribution of chemical functional groups on a nanoparticle because of the presence of multiple reactive sites on its surface. In cell-based analyses, nanoparticle multivalence can cause cross-linking of cell-surface proteins involved in signal pathway activation, and thus dramatically reduce receptor binding capability. In molecular analysis, multivalence can also lead to cross linking of molecules and reduce the sensitivity of the assay. Here, we propose to take advantage of the ability to synthesize polymers with pre-defined number of functional groups to form monovalent Pdots.
Fluorescence based techniques, from cellular imaging to bioassays, have become an indispensable tool kit in both basic cellular studies of cancer and in clinical applications and in vitro diagnostics. This project aims to develop a new class of robust and high-performance fluorescent probes for use in the molecular analysis and study of cancer cells. Successful completion of this project will result in a new reagent for use in the molecular analysis and cellular studies of cancer.
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|Chen, L; Wu, L; Yu, J et al. (2017) Highly photostable wide-dynamic-range pH sensitive semiconducting polymer dots enabled by dendronizing the near-IR emitters. Chem Sci 8:7236-7245|
|Yu, Jiangbo; Rong, Yu; Kuo, Chun-Ting et al. (2017) Recent Advances in the Development of Highly Luminescent Semiconducting Polymer Dots and Nanoparticles for Biological Imaging and Medicine. Anal Chem 89:42-56|
|Chen, Dandan; Wu, I-Che; Liu, Zhihe et al. (2017) Semiconducting polymer dots with bright narrow-band emission at 800 nm for biological applications. Chem Sci 8:3390-3398|
|Wu, Li; Wu, I-Che; DuFort, Christopher C et al. (2017) Photostable Ratiometric Pdot Probe for in Vitro and in Vivo Imaging of Hypochlorous Acid. J Am Chem Soc 139:6911-6918|
|Kuo, Chun-Ting; Peng, Hong-Shang; Rong, Yu et al. (2017) Optically Encoded Semiconducting Polymer Dots with Single-Wavelength Excitation for Barcoding and Tracking of Single Cells. Anal Chem 89:6232-6238|
|Wu, Xu; Wu, Li; Wu, I-Che et al. (2016) Copper (II)-doped semiconducting polymer dots for nitroxyl imaging in live cells. RSC Adv 6:103618-103621|
|Kuo, Chun-Ting; Thompson, Alison M; Gallina, Maria Elena et al. (2016) Optical painting and fluorescence activated sorting of single adherent cells labelled with photoswitchable Pdots. Nat Commun 7:11468|
|Ye, Fangmao; White, Collin C; Jin, Yuhui et al. (2015) Toxicity and oxidative stress induced by semiconducting polymer dots in RAW264.7 mouse macrophages. Nanoscale 7:10085-10093|
|Peng, Hong-Shang; Chiu, Daniel T (2015) Soft fluorescent nanomaterials for biological and biomedical imaging. Chem Soc Rev 44:4699-722|
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