Tip-enhanced fluorescence with radially polarized illumination for monitoring loop-mediated isothermal amplification
Shih-Chung Wei1, Tsung-Liang Chuang2, Da-Shin Wang2
1National Taiwan University, Institute of Biomedical Electronics and Bioinformatics, No. 1, Section 4, Roosevelt Road, Taipei 10617, Taiwan.
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.
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