Tip-enhanced fluorescence with radially polarized illumination for monitoring loop-mediated isothermal amplification on Hepatitis C virus cDNA
Journal
Journal of Biomedical Optics
Journal Volume
20
Journal Issue
2
Date Issued
2015-02
Author(s)
Shih-Chung Wei
Tsung-Liang Chuang
Da-Shin Wang
Hui-Hsin Lu
Frank X. Gu
Kung-Bin Sung
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/μ l 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. ? 2015 Society of Photo-Optical Instrumentation Engineers.
Subjects
Hepatitis C virus; loop-mediated isothermal amplification; radial polarized illumination; tip-enhanced fluorescence
SDGs
Other Subjects
Fluorescence; Viruses; DNA amplification; DNA replications; Enhanced fluorescence; Fluorescein isothiocyanate; Hepatitis C virus; Loop mediated isothermal amplifications; Polarized illumination; Radially polarized; Isotherms; Hepatitis C virus; complementary DNA; fluorescein; fluorescent dye; virus RNA; chemistry; devices; fluorescence microscopy; genetics; Hepacivirus; nanotechnology; nucleic acid amplification; procedures; DNA, Complementary; Fluorescein; Fluorescent Dyes; Hepacivirus; Microscopy, Fluorescence; Nanotechnology; Nucleic Acid Amplification Techniques; RNA, Viral
Type
journal article