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JHC exPRESS: First Published December 10, 2007. doi:10.1369/jhc.7A7313.2007
Copyright © Histochemical Society, Inc.

A more recent version of this article appeared on April 1, 2008.
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Articles

Raman Nanoparticle Probes for Antibody-based Protein Detection in Tissues

Barry Lutz 1, Claire Dentinger 1, Lei Sun 1, Lienchi Nguyen 1, Jingwu Zhang 1, A.J. Chmura 1, April Allen 1, Selena Chan 1 and Beatrice Knudsen 1*

1 Biomedical/Life Sciences, Digital Health Group, Intel Corporation, Santa Clara, California (BL,CD,LS,LN,JZ,AJC,SC), and Division of Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington (AA,BK)

* To whom correspondence should be addressed. E-mail: bknudsen{at}fhcrc.org.

Submitted on July 2, 2007
Accepted on 30 November 2007


  Abstract
Surface-enhanced Raman scattering (SERS) nanoparticles are emerging as a new approach for optical detection of biomolecules. In a model assay in formalin-fixed, paraffin-embedded (FFPE) prostate tissue sections, we detect prostate-specific antigen (PSA) using antibody (Ab) conjugated to Composite Organic-Inorganic Nanoparticles (COINs), and we use identical staining protocols to compare COIN-Ab and Alexa-Ab conjugates in adjacent tissue sections. Spectral analysis illustrates the fundamental difference between fluorescence and Raman signatures and accurately extracts COIN probe signals from background autofluorescence. Probe signals are used to generate images of PSA expression on the tissue, and quality measures are presented to characterize the performance of the COIN assay by comparison to Alexa. The staining accuracy (ability to correctly identify PSA expression in epithelial cells) is somewhat less for COIN than Alexa, which is attributed to an elevated false negative rate of the COIN. However, COIN provided signal intensities comparable to Alexa, and good intra-, inter- and lot-to-lot consistencies. Overall, COIN and Alexa detection reagents possess similar performance with FFPE tissues, supporting the further development of Raman probes for this application. This manuscript contains online supplemental material at http://www.jhc.org. Please visit this article online to view these materials.

Key Words: FFPE tissue, multiplex detection, surface-enhanced Raman scattering, autofluorescence, spectral analysis


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