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Proceedings Paper

Simultaneous molecular imaging of EGFR and HER2 using hyperspectral darkfield microscopy and immunotargeted nanoparticles
Author(s): Matthew Crow; Stella Marinakos; Ashutosh Chilkoti; Adam Wax
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Paper Abstract

Epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor (HER2) contribute to the regulation of cell proliferation, and when jointly over-expressed are associated with several types of cancer. The ability to monitor both receptors simultaneously results in a more accurate indicator of degree of cancerous activity than either receptor alone. Plasmonic nanoparticles (NPs) show promise as a potential EGFR and HER2 biomarker over alternatives such as fluorophores and quantum dots, which are limited by their cytotoxicity and photobleaching. To observe immunolabeled NPs bound to receptor-expressing cells, our past experiments were conducted using a novel optical darkfield microspectroscopy system. We implemented an epi-illumination darkfield broadband light train, which allows for darkfield analysis of live cells in culture with enhanced NP contrast. Under this setup, molecularly specific binding of NPs immunolabeled with anti-EGFR was confirmed. We have since adapted our darkfield setup, which previously only obtained spectral information from a line imaging spectrometer, to incorporate hyperspectral imaging capabilities, allowing widefield data acquisition within seconds. The new system has been validated through observation of shifts in the peak wavelength of scattering by gold NPs on silanated cover glasses using several immersion media. Peak resonant scattering wavelengths match well with that predicted by Mie theory. We will further demonstrate the potential of the system with simultaneous molecular imaging of multiple receptors in vitro using labeled EGFR+/HER2+ SK-BR-3 human breast cancer cells with anti-EGFR immunolabeled gold nanospheres and anti-HER2 immunolabeled gold nanorods, with each scattering in different spectral windows. Additional trials will be performed to demonstrate molecularly specific binding using EGFR+/HER2- MDA-MB-468 and HER2+/EGFR- MDA-MB-453 breast cancer cells.

Paper Details

Date Published: 17 February 2009
PDF: 7 pages
Proc. SPIE 7192, Plasmonics in Biology and Medicine VI, 71920P (17 February 2009); doi: 10.1117/12.808968
Show Author Affiliations
Matthew Crow, Duke Univ. (United States)
Stella Marinakos, Duke Univ. (United States)
Ashutosh Chilkoti, Duke Univ. (United States)
Adam Wax, Duke Univ. (United States)

Published in SPIE Proceedings Vol. 7192:
Plasmonics in Biology and Medicine VI
Tuan Vo-Dinh; Joseph R. Lakowicz, Editor(s)

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