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Bulletin of the Korean Chemical Society (BKCS)

ISSN 0253-2964(Print)
ISSN 1229-5949(Online)
Volume 34, Number 9
BKCSDE 34(9)
September 20, 2013 

Single C-Reactive Protein Molecule Detection on a Gold-Nanopatterned Chip Based on Total Internal Reflection Fluorescence
Yunmi Heo, Seungah Lee, Sang-Won Lee, Seong Ho Kang*
C-reactive protein, Gold nanoarray chip, Single-molecule detection, Total internal reflection fluorescence (TIRF)
Single C-reactive protein (CRP) molecules, which are non-specific acute phase markers and products of the innate immune system, were quantitatively detected on a gold-nanopatterned biochip using evanescent fieldenhanced fluorescence imaging. The 4 × 5 gold-nanopatterned biochip (spot diameter of 500 nm) was fabricated by electron beam nanolithography. Unlabeled CRP molecules in human serum were identified with single-molecule sandwich immunoassay by detecting secondary fluorescence generated by total internal reflection fluorescence (TIRF) microscopy. With decreased standard CRP concentrations, relative fluorescence intensities reduced in the range of 33.3 zM-800 pM. To enhance fluorescence intensities in TIRF images, the distance between biochip surface and CRP molecules was optimally adjusted by considering the quenching effect of gold and the evanescent field intensity. As a result, TIRF only detected one single-CRP molecule on the biochip the first time.
2725 - 2730
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