Plasmonic Labels Research Articles

A U-bent fiberoptic absorbance biosensor array (ArFAB) for multiplexed analyte detection

Compact and dip-type U-bent fiber optic sensor (U-FOS) probes exhibit remarkably high evanescent wave absorbance (EWA) sensitivity and are highly suitable for label-free multiplexed detection of proteins, virii and bacteria by exploiting their intrinsic optical absorption property in the UV region. Here, we show the design and development of a novel EWA based eight-channel array fiber optic absorbance biosensor (ArFAB) using U-FOS probes for multi-analyte detection or multi-sample analysis in real-time. A proof-of-concept ArFAB is designed to consist of a UV LED (λmax = 285 nm) and a highly sensitive CMOS linear image sensor coupled to a sensor probe module through fan-out (1-to-8) and parallel (8 nos) fiber bundles respectively. The sensor probe module is designed to hold and efficiently couple the light to/from 8 U-FOS probes with the help of ceramic ferrules and mating sleeves. The CMOS line sensor, with 8 coupling fibers equidistantly placed along its length, distinctly quantifies the intensity response from each of the eight U-FOS probes. The ArFAB was systematically validated for reproducible optical intensity measurements and label-free multiplexed detection of proteins and bacteria was realized. Moreover, a novel sandwich immunoassay with plasmonic labels for immunoglobulin G (IgG) with picomolar analyte detection limits at 280 nm wavelength is demonstrated. With mass-producible U-FOS probes and a low-cost optoelectronic instrumentation that can be easily customized for LSPR or any other sensing phenomenon, the ArFAB is highly promising for cost-effective biomolecular interactions, and cell analyses and clinical diagnostic applications.

(Invited) Development of New Strategies for Bringing Photoelectrochemical Biosensing to the Point-of-Need

Photoelectrochemistry combines light excitation with electrochemical readout for lowering the bias voltage needed for performing electrochemical reactions. As a result, when used in biosensing, photoelectrochemical signal readout reduces the background signals, lowering the limit-of-detection of such biosensors. To enable photoelectrochemical (PEC) signal readout to be applied to point-of-need biosensing, we have taken a three tiered approach focused on improving the understanding of signal transduction in PEC Biosensing, developing label-free assays, and creating handheld readout platforms.In this work, we developed a system using DNA as a nano-ruler to control the distance between plasmonic nanoparticles and PEC electrodes. This system was used to rationally-design PEC material systems for signal-on biosensing. Using this materials architecture, we developed a signal-on biosensor without target labeling for detecting DNA hybridization. This assay uses sequential DNA hybridization to generate a PEC signal. First, the DNA target is captured on probe-modified photoelectrodes. This is followed by hybridization of the unbound probes with DNA strands modified with plasmonic labels. The plasmonic label modulates the PEC signal, increasing the measured PEC current at low target concentrations. To enable biosensing at the point-of-need, we also developed a handheld PEC reader. The integration of plasmonic nanoparticles with PEC electrodes, label-free DNA assays, and handheld PEC readout paves the way toward bringing point-of-need PEC Biosensing.

Plasmonic Fiberoptic Absorbance Biosensor (P-FAB) for Rapid Detection of SARS-CoV-2 Nucleocapsid Protein.

SARS-CoV-2 nucleocapsid protein-based COVID-19 diagnosis is a promising alternative to the high-priced, time-consuming, and labor-intensive RT-PCR tests. Here, we developed a rapid, dip-type, wash-free plasmonic fiber optic absorbance biosensor (P-FAB) strategy for the point-of-care detection of SARS-CoV-2 N-protein, expressed abundantly during the infection. P-FAB involves a sandwich assay with plasmonic labels on the surface of a U-bent fiber optic sensor probe with a high evanescent wave absorbance (EWA) sensitivity. The SARS-CoV-2 N-protein is quantified in terms of the change in the intensity of the light propagating through the U-bent sensor probe coupled to a green LED and a photodetector. Firstly, the optical fiber material (silica vs. polymeric optical fiber), was evaluated to realize a sensitive sensor platform. The optimal size of AuNP labels (20, 40, and 60 nm) to achieve high sensitivity and a lower limit of detection (LoD) was investigated. Following the P-FAB strategy, fused silica/glass optical fiber (GOF) U-bent senor probe and citrate-capped AuNP labels (size ~40 nm) gave rise to an LoD down to ~2.5 ng/mL within 10 mins of read-out time. Further, studies on development and validation of a point of care (PoC) read-out device, and preclinical studies are in progress.

Development of a U-bent plastic optical fiber biosensor with plasmonic labels for the detection of chikungunya non-structural protein 3.

This study presents a novel plasmonic fiber optic sandwich immunobiosensor for the detection of chikungunya, an infectious mosquito-borne disease with chronic musculoskeletal pain and acute febrile illness, by exploiting non-structural protein 3 (CHIKV-nsP3) as a biomarker. A plasmonic sandwich immunoassay for CHIKV-nsP3 was realized on the surface of a compact U-bent plastic optical fiber (POF, 0.5 mm core diameter) with gold nanoparticles (AuNPs) as labels. The high evanescent wave absorbance (EWA) sensitivity of the U-bent probes allows the absorption of the light passing through the fiber by the AuNP labels, upon the formation of a sandwich immunocomplex of CHIKV-nsP3 on the core surface of the U-bent probe region. A simple optical set-up with a low-cost green LED and a photodetector on either end of the U-bent probe gave rise to a detection limit of 0.52 ng mL-1 (8.6 pM), and a linear range of 1-104 ng mL-1 with a sensitivity of 0.1043A530 nm/log(CnsP3). In addition, the plasmonic POF biosensor shows strong specificity towards the CHIKV-nsP3 analyte in comparison with Pf-HRP2, HIgG, and dengue whole virus. The results illustrate the potential of plasmonic POF biosensors for direct and sensitive point-of-care detection of the chikungunya viral disease.

SPRi-Based Biosensing Platforms for Detection of Specific DNA Sequences Using Thiolate and Dithiocarbamate Assemblies.

The framework of presented study covers the development and examination of the analytical performance of surface plasmon resonance-based (SPR) DNA biosensors dedicated for a detection of model target oligonucleotide sequence. For this aim, various strategies of immobilization of DNA probes on gold transducers were tested. Besides the typical approaches: chemisorption of thiolated ssDNA (DNA-thiol) and physisorption of non-functionalized oligonucleotides, relatively new method based on chemisorption of dithiocarbamate-functionalized ssDNA (DNA-DTC) was applied for the first time for preparation of DNA-based SPR biosensor. The special emphasis was put on the correlation between the method of DNA immobilization and the composition of obtained receptor layer. The carried out studies focused on the examination of the capability of developed receptors layers to interact with both target DNA and DNA-functionalized AuNPs. It was found, that the detection limit of target DNA sequence (27 nb length) depends on the strategy of probe immobilization and backfilling method, and in the best case it amounted to 0.66 nM. Moreover, the application of ssDNA-functionalized gold nanoparticles (AuNPs) as plasmonic labels for secondary enhancement of SPR response is presented. The influence of spatial organization and surface density of a receptor layer on the ability to interact with DNA-functionalized AuNPs is discussed. Due to the best compatibility of receptors immobilized via DTC chemisorption: 1.47 ± 0.4 · 1012 molecules · cm−2 (with the calculated area occupied by single nanoparticle label of ~132.7 nm2), DNA chemisorption based on DTCs is pointed as especially promising for DNA biosensors utilizing indirect detection in competitive assays.

Ultrasensitive detection of HIV-1 p24 antigen by a hybrid nanomechanical-optoplasmonic platform with potential for detecting HIV-1 at first week after infection.

Early detection of HIV infection is the best way to prevent spread of the disease and to improve the efficiency of the antiretroviral therapy. Nucleic acid amplification tests (NAAT) have become the gold-standard for detecting low-concentrations of the virus in blood. However, these methods are technically demanding and cost-prohibitive in developing countries. Immunoassays are more affordable and can be more easily adapted for point-of-care diagnosis. However, the sensitivity so far of these methods has been too low. We here report the development of a sandwich immunoassay that combines nanomechanical and optoplasmonic transduction methods for detecting the HIV-1 capsid antigen p24 in human serum. The immunoreactions take place on the surface of a compliant microcantilever where gold nanoparticles are used as both mechanical and plasmonic labels. The microcantilever acts as both a mechanical resonator and an optical cavity for the transduction of the mechanical and plasmonic signals. The limit of detection of the immunoassay is 10−17 g/mL that is equivalent to one virion in 10 mL of plasma. This is 5 orders of magnitude better than last generation of approved immunoassays and 2 orders of magnitude better than NAAT. This technology meets the demands to be produced en masse at low cost and the capability for miniaturization to be used at the point-of-care.

Shape-changing magnetic assemblies as high-sensitivity NMR-readable nanoprobes.

Fluorescent and plasmonic labels and sensors have revolutionized molecular biology, helping visualize in vitro cellular and biomolecular processes1–3. Increasingly, such probes are now designed to respond to wavelengths in the near infrared region, where reduced tissue autofluorescence and photon attenuation enable subsurface in vivo sensing4. But even in the near infrared, optical resolution and sensitivity decrease rapidly with increasing depth. Here we present a sensor design that obviates the need for optical addressability by operating in the NMR radio-frequency (RF) spectrum, where signal attenuation and distortion by tissue and biological media are negligible, where background interferences vanish, and where sensors can be spatially located using standard magnetic resonance imaging (MRI) equipment. The RF-addressable sensor assemblies presented here are comprised of pairs of magnetic disks spaced by swellable hydrogel material; they reversibly reconfigure in rapid response to chosen stimuli, to give geometry-dependent, dynamic NMR spectral signatures. Sensors can be made from biocompatible materials, are detectable down to low concentrations, and offer potential responsive NMR spectral shifts approaching a million times those of traditional magnetic resonance spectroscopies. Inherent adaptability should allow such shape-changing systems to measure numerous different environmental and physiological indicators, affording broadly generalizable, MRI-compatible, RF analogues to optically-based probes for use in basic chemical, biological and medical research.

Direct immunosensing by spectral correlation interferometry: assay characteristics versus antibody immobilization chemistry.

A 3-channel biosensor based on spectral correlation interferometry (SCI) has been adapted for direct optical detection of antigens by measuring changes in thickness of a biolayer on functionalized glass slips employed as affordable single-use sensor chips. The instrument is insensitive to the bulk refractive index of a solution under test and provides signals in metrological units (pm or nm). Using real-time monitoring with the SCI, protocols for fabrication of sensor chips with different functional (epoxylated, carboxylated, and biotinylated) surfaces for antibody immobilization have been developed and optimized to minimize chip-to-chip variations and achieve better limit of detection (LOD), shorter assay time, and longer shelf life. The optimized coupling surfaces have been compared for detection of human serum albumin (HSA) used as a model agent of medical significance. The dynamic ranges for measuring the HSA concentration were 0.07-20, 0.12-30, and 0.25-10 μg/ml, and the assay durations were less than 20, 15, and 30 min for the epoxylated, carboxylated, and biotinylated chips, respectively. The advantages of each type of sensor chip have been shown, namely, the carboxylated chips feature the shortest assay time, the epoxylated ones demonstrate the best LOD, and the biotinylated chips exhibit the longest shelf life in an unprotected environment. The developed protocols of antibody immobilization can be used in different biosensors and assay techniques including those based on fluorescent, magnetic or plasmonic labels, etc. The SCI is well compatible with various partially transparent layers used in biosensing and with microarrays for multi-analyte detection.