Won Jung Yoon

A regenerative label-free fiber optic sensor using surface plasmon resonance for clinical diagnosis of fibrinogen

Purpose We present the regenerative label-free fiber optical biosensor that exploits surface plasmon resonance for quantitative detection of fibrinogen (Fbg) extracted from human blood plasma. Materials and methods The sensor head was made up of a multimode optical fiber with its polymer cladding replaced by metal composite of nanometer thickness made of silver, aluminum, and nickel. The Ni layer coated allowed a direct immobilization of histidine-tagged peptide (HP) on its metal surface without an additional cross-linker in between. On the coated HP layer, immunoglobulin G was then immobilized for specific capturing of Fbg. Results We demonstrated a real-time quantitative detection of Fbg concentrations with limit of detection of ~10 ng/mL. The fact that the HP layer could be removed by imidazole with acid also permitted us to demonstrate the regeneration of the outermost metal surface of the sensor head for the sensor reusability. Conclusion The sensor detection limit was estimated to be ~10 pM, which was believed to be sensitive enough for detecting Fbg during the clinical diagnosis of cardiovascular diseases, myocardial infarction, strokes, and Alzheimer’s diseases.

Dielectric Metal-Based Multilayers for Surface Plasmon Resonance with Enhanced Quality Factor of the Plasmonic Waves

We present improved quality factors of surface plasmon resonance (SPR) in a prism-based Krätschman configuration by using double the number of metal–dielectric layers for extended long-range surface plasmon. These multilayers lead to the coupling of multi-plasmonic waves for enhanced depth-to-width ratio (Γ) of the SPR dip of the reflectance curve. We use a transfer matrix approach to numerically simulate the curve of reflectance versus incident angle with each layer thickness optimized. We find that the four layers comprising doubled Teflon-Ag multilayers produce Γ higher than a single layer of Ag by a factor of about 122. These enhanced Γ (related to enhanced quality factor of the SPR wave) that lead to enlarged depth of SPR evanescent field penetration, can readily find applications in fluorescence detection with its efficiency elevated, which is required for fluorescence-based assays where weak fluorescent signals are expected, such as biological diagnosis that uses small volumes of liquid containing fluorescent dyes.

Reproducible Enhancement of Fluorescence by Bimetal Mediated Surface Plasmon Coupled Emission for Highly Sensitive Quantitative Diagnosis of Double‐Stranded DNA

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-1 (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.

Blood-based immunoassay of tau proteins for early diagnosis of Alzheimer's disease using surface plasmon resonance fiber sensors

We present the immunoassay of tau proteins (total tau and phosphorylated tau) in human sera using surface plasmon resonance (SPR) fiber sensors. This assay aimed at harvesting the advantages of using both SPR fiber sensors and a blood-based assay to demonstrate label-free point-of-care-testing (POCT) patient-friendly assay in a compact format for the early diagnosis of Alzheimers disease (AD). For conducting the assay, we used human sera of 40 subjects divided into halves, which were grouped into AD patients and control groups according to a number of neuropsychological tests. We found that on an average, the concentrations of both total tau and phosphorylated tau proteins (all known to be higher in cerebrospinal fluid (CSF) and the brain) turned out to be higher in human sera of AD patients than in controls. The limits of detection of total tau and phosphorylated tau proteins were 2.4 pg mL−1 and 1.6 pg mL−1, respectively. In particular, it was found that the AD group exhibited average concentration of total tau proteins 6-fold higher than the control group, while concentration of phosphorylated tau proteins was 3-fold higher than that of the control. We can attribute this inhomogeneity between both types of tau proteins (in terms of increase of control-to-AD in average concentration) to un-phosphorylated tau proteins being more likely to be produced in blood than phosphorylated tau proteins, which possibly is one of the potential key elements playing an important role in AD progress.

A Plasmonic Fiber Based Glucometer and Its Temperature Dependence

We present the plasmonic fiber based optical glucometer. A thin gold layer is coated on clad-free core of multimode optical fiber along 3 cm length to excite surface plasmons at 632.8 nm wavelength. Glucose oxidase is immobilized on the metal surface for glucose sensing. The effective surface refractive index increases by gluconic acid and hydrogen peroxide that are generated upon glucose injection, leading to plasmonic condition change with a consequence of optical power change at the fiber output. We obtain limit of detection of glucose concentration of 6.75 mg/dL, indicating higher sensitivity than the wavelength interrogating SPR glucometer that uses a spectrometer of 1nm spectral resolution. The coefficient of variation is 8.6% at a glucose concentration of 80 mg/dL at room temperature. We also examine the effects of ambient temperature variations from −10 °C to 40 °C on the performance of the presented sensor and compared them with those on commercially available glucometers that are based on enzyme electrodes. We find that the presented fiber sensor produced standard deviation of 12.1 mg/dL at a glucose concentration of 80 mg/dL under such varying temperature, which is, even without additional temperature correction function, comparable to the commercialized ones.

Liquid Cladding Mediated Optical Fiber Sensors for Copper Ion Detection

We present a label-free optical fiber based sensor device to detect copper ions (Cu2+) in water. A multimode optical fiber, with its polymer cladding removed along a 1-cm length, is used for the optical sensor head, where the injected Cu2+ in the liquid phase acts as a liquid cladding for the optical mode. The various Cu2+ concentrations modulate the numerical aperture (NA) of the liquid cladding waveguide part. The degree of NA mismatch between the liquid cladding and solid cladding guided parts gives rise to an optical power transmittance change, forming the sensing principle. The presented liquid cladding fiber sensor exhibits a minimum resolvable refractive index of 2.48 × 10−6. For Cu2+ detection, we functionalize the sensor head surface (fiber core) using chitosan conjugated ethylenediaminetetraacetic acid (EDTA) which captures Cu2+ effectively due to the enhanced chelating effects. We obtain a limit of detection of Cu2+ of 1.62 nM (104 ppt), which is significantly lower than the tolerable level in drinking water (~30 µM), and achieve a dynamic range of 1 mM. The simple structure of the sensor head and the sensing system ensures the potential capability of being miniaturized. This may allow for in-situ, highly-sensitive, heavy metal sensors in a compact format.

Fluorescence Enhancement Using Bimetal Surface Plasmon-Coupled Emission from 5-Carboxyfluorescein (FAM)

We demonstrate the enhancement of fluorescence emission from a dye, 5-carboxyfluorescein (FAM), which couples with surface plasmons at the spectral channels of excitation and emission. Experiments and calculations revealed that bimetallic (gold-silver) plasmon, as compared to the monometallic ones, allowed such coupling to be enhanced, at both the spectral channels. We achieved the maximum fluorescence enhancement level of 46.5-fold, with markedly high reproducibility (coefficient of variation ~ 0.5%) at a FAM concentration of 10 nM. We also found that higher fluorescence enhancement was more likely to be reproducible. This encourages the use of this technology for practical applications in fluorescence-based biochemical assays. Moreover, we investigated a FAM concentration-dependent enhancement of fluorescence. It was found that fluorescence enhancement decreased and saturated at above 10 nM concentration possibly due to partial photo-bleaching of FAM molecules.