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Developing luminescent nanoprobes for labeling focal adhesion complex proteins and performing combined AFM-TIRF imaging of these conjugates

Recent progress in the field of semiconductor nanocrystals or Quantum Dots (QDs)
has seen them find wider acceptance as a tool in biomedical research labs. As produced,
high quality QDs synthesized by high temperature organometallic synthesis, are coated
with a hydrophobic ligand. Therefore, they must be further processed to be soluble in
water and made biocompatible.
A process to coat the QDs with silk fibroin, a fibrous protein derived from the
Bombyx mori silk worm, is described. Following the coating process, the characterization
of size, optical properties and biocompatibility profile of these particle systems is
described. In addition, conjugation of the silk fibroin coated QDs to different labeling
proteins such as phalloidin and streptavidin is described.
Proteins on the surface of ovarian cancer cells (HeyA8) and of cytoskeletal
components participating in the formation of focal adhesion complex (FAC), such as F-actin
in endothelial cells (HUVECS) were labeled using the bio-conjugated QDs. Various imaging techniques such as epi-fluorescence, TIRF and AFM were used to
study the QD labeled cells. Overall the project has produced luminescent nanoprobes that
enable the study of FAC formation dynamics and potentially a better in vivo fluorescent
marker tool.

Identiferoai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/85904
Date10 October 2008
CreatorsNathwani, Bhavik Bharat
ContributorsMeissner, Kenith E.
PublisherTexas A&M University
Source SetsTexas A and M University
Languageen_US
Detected LanguageEnglish
TypeBook, Thesis, Electronic Thesis, text
Formatelectronic, born digital

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