Plasmonic enhancement of fluorescence from SYBR Green I conjugated with a double-stranded DNA (dsDNA) amplicon is demonstrated on polymerase chain reaction (PCR) products. Theoretical computation leads to use of the bimetallic (Au 2 nm-Ag 50 nm) surface plasmons due to larger local fields (higher quality factors) than monometallic (Ag or Au) ones at both dye excitation and emission wavelengths simultaneously, optimizing fluorescence enhancement with surface plasmon coupled emission (SPCE). Two kinds of reverse Kretschmann configurations are used, which favor, in signal-to-noise ratio, a fluorescence assay that uses optically dense buffer such as blood plasma. The fluorescence enhancement (12.9 fold at maximum) with remarkably high reproducibility (coefficient of variation (CV) < 1%) is experimentally demonstrated. This facilitates credible quantitation of enhanced fluorescence, however unlikely to obtain by localized surface plasmons. The plasmon-induced optical gain of 46 dB due to SPCE-active dye molecules is also estimated. The fluorescence enhancement technologies with PCR enables LOD of the dsDNA template concentration of ≈400 fg µL (CV < 1%), the lowest ever reported in DNA fluorescence assay to date. SPCE also reduces photobleaching significantly. These technologies can be extended for a highly reproducible and sufficiently sensitive fluorescence assay with small volumes of analytes in multiplexed diagnostics.

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We demonstrated modulation of the waveguide mode mismatch via liquid cladding of the controllable refractive index for label-free quantitative detection of concentration of chemical or biological substances. A multimode optical fiber with its core exposed was used as the sensor head with the suitable chemical modification of its surface. Injected analyte liquid itself formed the liquid cladding for the waveguide. We found that modulation of the concentration of analyte injected enables a degree of the waveguide mode mismatch to be controlled, resulting in sensitive change in optical power transmission, which was utilized for its real-time quantitative assay. We applied the device to quantitating concentration of glycerol and bovine serum albumin (BSA) solutions. We obtained experimentally the limit of detection (LOD) of glycerol concentration, 0.001% (volume ratio), corresponding to the resolvable index resolution of ∼1.02 × 10 RIU (refractive index unit). The presented sensors also exhibited reasonably good reproducibility. In BSA detection, the sensor device response was sensitive to change in the refractive indices not only of liquid bulk but also of layers just above the sensing surface with higher sensitivity, providing the LOD experimentally as ∼3.7 ng/mL (mass coverage of ∼30 pg/mm). A theoretical model was also presented to invoke both mode mismatch modulation and evanescent field absorption for understanding of the transmission change, offering a theoretical background for designing the sensor head structure for a given analyte. Interestingly, the device sensing length played little role in the important sensor characteristics such as sensitivity, unlike most of the waveguide-based sensors. This unraveled the possibility of realizing a highly simple structured label-free sensor for point-of-care testing in a real-time manner via an optical waveguide with liquid cladding. This required neither metal nor dielectric coating but still produced sensitivity comparable to those of other types of label-free sensors such as plasmonic fiber ones.