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

Shih-Chung Wei; Tsung-Liang Chuang; Da-Shin Wang; Hui-Hsin Lu; Frank X Gu; Kung-Bin Sung; Chii-Wann Lin

doi:10.1117/1.JBO.20.2.027005

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

J Biomed Opt. 2015 Feb;20(2):27005. doi: 10.1117/1.JBO.20.2.027005.

Authors

Shih-Chung Wei¹, Tsung-Liang Chuang², Da-Shin Wang², Hui-Hsin Lu², Frank X Gu³, Kung-Bin Sung⁴, Chii-Wann Lin⁵

Affiliations

¹ National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.
² National Taiwan University, Institute of Biomedical Engineering, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.
³ University of Waterloo, Department of Chemical Engineering, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
⁴ National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, TaiwandNational Taiwan University, Molecular Imaging Center, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.
⁵ National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, TaiwanbNational Taiwan University, Institute of Biomedical Engineering, No. 1, Section 4, Roosevelt Road, Taipei 10617, Tai.

PMID: 25679746
DOI: 10.1117/1.JBO.20.2.027005

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 0.004 ng∕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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

DNA, Complementary / analysis*
DNA, Complementary / genetics
Fluorescein / chemistry
Fluorescent Dyes / chemistry*
Hepacivirus / genetics*
Microscopy, Fluorescence
Nanotechnology
Nucleic Acid Amplification Techniques / instrumentation
Nucleic Acid Amplification Techniques / methods*
RNA, Viral / analysis
RNA, Viral / genetics

Substances

DNA, Complementary
Fluorescent Dyes
RNA, Viral
Fluorescein