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Proceedings Paper

Whole-body multicolor spectrally resolved fluorescence imaging for development of target-specific optical contrast agents using genetically engineered probes
Author(s): Hisataka Kobayashi; Yukihiro Hama; Yoshinori Koyama; Tristan Barrett; Yasuteru Urano; Peter L. Choyke
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Paper Abstract

Target-specific contrast agents are being developed for the molecular imaging of cancer. Optically detectable target-specific agents are promising for clinical applications because of their high sensitivity and specificity. Pre clinical testing is needed, however, to validate the actual sensitivity and specificity of these agents in animal models, and involves both conventional histology and immunohistochemistry, which requires large numbers of animals and samples with costly handling. However, a superior validation tool takes advantage of genetic engineering technology whereby cell lines are transfected with genes that induce the target cell to produce fluorescent proteins with characteristic emission spectra thus, identifying them as cancer cells. Multicolor fluorescence imaging of these genetically engineered probes can provide rapid validation of newly developed exogenous probes that fluoresce at different wavelengths. For example, the plasmid containing the gene encoding red fluorescent protein (RFP) was transfected into cell lines previously developed to either express or not-express specific cell surface receptors. Various antibody-based or receptor ligand-based optical contrast agents with either green or near infrared fluorophores were developed to concurrently target and validate cancer cells and their positive and negative controls, such as &bgr;-D-galactose receptor, HER1 and HER2 in a single animal/organ. Spectrally resolved fluorescence multicolor imaging was used to detect separate fluorescent emission spectra from the exogenous agents and RFP. Therefore, using this in vivo imaging technique, we were able to demonstrate the sensitivity and specificity of the target-specific optical contrast agents, thus reducing the number of animals needed to conduct these experiments.

Paper Details

Date Published: 14 February 2007
PDF: 10 pages
Proc. SPIE 6449, Genetically Engineered and Optical Probes for Biomedical Applications IV, 644914 (14 February 2007); doi: 10.1117/12.724241
Show Author Affiliations
Hisataka Kobayashi, National Cancer Institute, National Institutes of Health (United States)
Yukihiro Hama, National Cancer Institute, National Institutes of Health (United States)
Yoshinori Koyama, National Cancer Institute, National Institutes of Health (United States)
Tristan Barrett, National Cancer Institute, National Institutes of Health (United States)
Yasuteru Urano, The Univ. of Tokyo (Japan)
Peter L. Choyke, National Cancer Institute, National Institutes of Health (United States)


Published in SPIE Proceedings Vol. 6449:
Genetically Engineered and Optical Probes for Biomedical Applications IV
Samuel Achilefu; Darryl J. Bornhop; Ramesh Raghavachari; Alexander Pavlovich Savitsky; Rebekka M. Wachter, Editor(s)

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