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CLC number: TH742.1

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Received: 2005-01-28

Revision Accepted: 2005-04-05

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Journal of Zhejiang University SCIENCE A 2005 Vol.6 No.7 P.623~626

http://doi.org/10.1631/jzus.2005.A0623


Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence


Author(s):  WANG Li-qiang, LU Zu-kang

Affiliation(s):  State Key Lab of Modern Optical Instrumentation, Zhejiang University, Hangzhou 310027, China

Corresponding email(s):   wangliqiang@zju.edu.cn

Key Words:  Protein microarray, Evanescent excitation, Sensitivity, Total reflection


WANG Li-qiang, LU Zu-kang. Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence[J]. Journal of Zhejiang University Science A, 2005, 6(7): 623~626.

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author="WANG Li-qiang, LU Zu-kang",
journal="Journal of Zhejiang University Science A",
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pages="623~626",
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publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.A0623"
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%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.A0623

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T1 - Improving the sensitivity of protein microarray by evanescent-field-induced fluorescence
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DOI - 10.1631/jzus.2005.A0623


Abstract: 
To improve the sensitivity of protein microarray, a prism surface replaces the surface of the common microscope slide. The protein targets arrayed on the surface are hybridized and labelled by fluorescent probes. evanescent excitation occurs when the convergent laser reaches the surface, and a photomultiplier tube detects the emitted fluorescent signal. A two-dimensional actuator scans the whole surface to achieve planar laser excitation and fluorescence collection. The penetration depth of the evanescent field into the protein targets is only some hundred nanometers and can be controlled by different incident angle of the laser beam, so the undesired background signals are reduced dramatically and the detection sensitivity is improved by a factor of 50 to 100 comparing to confocal excitation. This approach can detect low abundance analytes without signal amplification.

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

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[2] Miklos, G.L., Maleszka, R., 2001. Protein functions and biological contexts. Proteomics, 1(2):169-178.

[3] Pawlak, M., Schick, E., Bopp, M.A., Schneider, M.J., Oroszlan, P., Ehrat, M., 2002. Zeptosens’ protein microarrays: a novel high performance microarray platform for low abundance protein analysis. Proteomics, 2(4):383-393.

[4] Rachlin, D.J., 2002. Optimized approach for microarray scanning. SPIE, 4623:13-26.

[5] Schena, M., 1999. DNA Microarrays−A Practical Approach. Oxford University Press, New York, p.17-42.

[6] Toomre, D., Manstein, D.J., 2001. Lighting up the cell surface with evanescent wave microscopy. TRENDS in Cell Biology, 11(7):298-302.

[7] Wang, L., Ni, X., Lu, Z., Zheng, X., Li, Y., 2004. Enhancing the quality metric of protein microarray image. Journal of Zhejiang University SCIENCE, 5(12):1621-1628.

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