Research Papers: Sensing

Tip-enhanced fluorescence with radially polarized illumination for monitoring loop-mediated isothermal amplification on Hepatitis C virus cDNA

[+] Author Affiliations
Shih-Chung Wei

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

Tsung-Liang Chuang, Da-Shin Wang, Hui-Hsin Lu

National Taiwan University, Institute of Biomedical Engineering, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

Frank X. Gu

University of Waterloo, Department of Chemical Engineering, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada

Kung-Bin Sung

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

National Taiwan University, Molecular Imaging Center, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

Chii-Wann Lin

National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

National Taiwan University, Institute of Biomedical Engineering, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan

J. Biomed. Opt. 20(2), 027005 (Feb 13, 2015). doi:10.1117/1.JBO.20.2.027005
History: Received November 2, 2014; Accepted January 12, 2015
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Abstract.  A tip nanobiosensor for monitoring DNA replication was presented. The effects of excitation power and polarization on tip-enhanced fluorescence (TEF) were assessed with the tip immersed in fluorescein isothiocyanate solution first. The photon count rose on average fivefold with radially polarized illumination at 50 mW. We then used polymerase-functionalized tips for monitoring loop-mediated isothermal amplification on Hepatitis C virus cDNA. The amplicon-SYBR® Green I complex was detected and compared to real-time loop-mediated isothermal amplification. The signals of the reaction using 4 and 0.004ng/μl templates were detected 10 and 30 min earlier, respectively. The results showed the potential of TEF in developing a nanobiosensor for real-time DNA amplification.

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© 2015 Society of Photo-Optical Instrumentation Engineers

Citation

Shih-Chung Wei ; Tsung-Liang Chuang ; Da-Shin Wang ; Hui-Hsin Lu ; Frank X. Gu, et al.
"Tip-enhanced fluorescence with radially polarized illumination for monitoring loop-mediated isothermal amplification on Hepatitis C virus cDNA", J. Biomed. Opt. 20(2), 027005 (Feb 13, 2015). ; http://dx.doi.org/10.1117/1.JBO.20.2.027005


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