Implementation and verification of the nanobiosensor based on spatially confined surface plasmon and metal enhanced fluorescence
Date Issued
2015
Date
2015
Author(s)
Wei, Shih-Chung
Abstract
In this manuscript, we used radially-polarized illumination to produce a strong and well-confined optical field at nanostructure to enhance fluorescence signal for biosensing. With the illumination intensity of 50 mW, fluorescein isothiocyanate emission was increased 5 times on average by tip-enhanced fluorescence (TEF). Moreover, we monitored the loop-mediated isothermal amplification (LAMP) on Hepatitis C virus cDNA (complementary DNA) by TEF (tip-LAMP). The increment of amplicon-SYBR at three different template concentrations was found 10-30 minutes earlier than general LAMP. For further increasing of the detection specificity, multiplex detection capability and preventing cross contamination, we have designed a fluorescence resonance energy transfer based loop-mediated isothermal amplification (FRET-LAMP) for Hepatitis C and B virus. The template of 10-3 and 10-4 g per 25l was easily detected in forty minutes by FRET-LAMP, while the reaction performed with SYBR green needs fifty to sixty minutes for 10-3 g per 25 l template. Furthermore, from single point to massive parallel reaction, FRET-LAMP at nanoarray was performed (Digital-LAMP). Digitizing counting was used to monitor the reaction. About 20 min earlier detection time was shown in FRET-LAMP nanoarray comparing to real-time LAMP. In addition, the confined optical field with radially-polarized illumination was also adopted to scanning surface plasmon microscopy (SSPM) to image the refractive index change of stacking layer on Au film. The characteristics of SSPM, lateral resolution and refractive index sensitivity, was inspected. SSPM were then applied to image Hepatitis C virus primer labeled nanoarray, 1, 2-Dioleoyl-sn-glycero-3-phosphocholine array, deposited graphene, placenta growth factor nanobiosensor.
Subjects
tip-enhanced fluorescence
surface plasmon microscopy
nanoarray
loop-mediated isothermal amplification
Hepatitis C virus
digitized counting
SDGs
Type
thesis
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