Optical Biosensing Platform Research Articles

Graphene oxide functionalized long period fiber grating for highly sensitive hemoglobin detection

We present graphene oxide (GO) nanosheets functionalized long period grating (LPG) for ultrasensitive hemoglobin sensing. The sensing mechanism relies on the measurement of LPG resonant intensity change induced by the adsorption of hemoglobin molecules onto GO, where GO as a bio-interface linkage provides the significant light-matter interaction between evanescent field and target molecules. The deposition technique based on chemical-bonding associated with physical-adsorption was developed to immobilize GO nanosheets on cylindrical fiber device. The surface morphology was characterized by scanning electron microscope, atomic force microscopy, and Raman spectroscopy. With relatively thicker GO coating, the refractive index (RI) sensitivity of GO-LPG was extremely enhanced and achieved −76.5 dB/RIU, −234.2 dB/RIU and +1580.5 dB/RIU for RI region of 1.33–1.38, 1.40–1.44 and 1.45–1.46, respectively. The GO-LPG was subsequently implemented as an optical biosensor to detect human hemoglobin giving a sensitivity of 1.9 dB/(mg/mL) and a detectable concentration of 0.05 mg/mL, which was far below the hemoglobin threshold value for anemia defined by World Health Organization. The proposed GO-LPG architecture can be further developed as an optical biosensing platform for anemia diagnostics and biomedical applications.

Fabrication of ordered metallic glass nanotube arrays for label-free biosensing with diffractive reflectance

In this study, a photoresist template with well-defined contact hole array was fabricated, to which radio frequency magnetron sputtering process was then applied to deposit an alloyed Zr55Cu30Al10Ni5 target, and finally resulted in ordered metallic glass nanotube (MGNT) arrays after removal of the photoresist template. The thickness of the MGNT walls increased from 98 to 126nm upon increasing the deposition time from 225 to 675s. The wall thickness of the MGNT arrays also increased while the dimensions of MGNT reduced under the same deposition condition. The MGNT could be filled with biomacromolecules to change the effective refractive index. The air fraction of the medium layer were evaluated through static water contact angle measurements and, thereby, the effective refractive indices the transverse magnetic (TM) and transverse electric (TE) polarized modes were calculated. A standard biotin–streptavidin affinity model was tested using the MGNT arrays and the fundamental response of the system was investigated. Results show that filling the MGNT with streptavidin altered the effective refractive index of the layer, the angle of reflectance and color changes identified by an L*a*b* color space and color circle on an a*b* chromaticity diagram. The limit of detection (LOD) of the MGNT arrays for detection of streptavidin was estimated as 25nM, with a detection time of 10min. Thus, the MGNT arrays may be used as a versatile platform for high-sensitive label-free optical biosensing.

An optical waveguide sensor based on mesoporous silica films with a comparison to surface plasmon resonance sensors

Mesoporous silica films (MSF) with large surface areas and high stability were fabricated by a simple Stöber solution growth method. The sensitivity and limit of detection (LOD) for an optical waveguide transverse electric mode (TE-OWG) sensor are 62.869 deg/RIU and 5.8250 × 10−6 RIU, respectively. The sensitivity and LOD for a surface plasma resonance (SPR) sensor are 131.58 deg/RIU and 1.0551 × 10−5 RIU, respectively. The MSF sensor can detect both small and large molecules. In the detection of non-adsorbing glucose, a LOD of 40.416 mg/L was recorded, which resulted from the change of the refractive index of the solution. For the detection of CTAB, which may adsorb onto the film, the LOD significantly decreases to 1.6315 nM. Although, large molecules, such as BSA, cannot diffuse into the mesoporous structure of MSF to adsorb onto the inner pore walls, physical adsorption on the outer surface of MSF still leads to a LOD of 84.209 pM. Fresnel calculations suggested higher potential sensitivity of the current system compared to that of the conventional SPR sensors. Thus, the MSF may be used as a versatile platform for high-sensitive label-free optical biosensing due to the high performance and the large potential of the surface functionality.

A label-free nanostructured plasmonic biosensor based on Blu-ray discs with integrated microfluidics for sensitive biodetection

Nanostructure-based plasmonic biosensors have quickly positioned themselves as interesting candidates for the design of portable optical biosensor platforms considering the potential benefits they can offer in integration, miniaturization, multiplexing, and real-time label-free detection. We have developed a simple integrated nanoplasmonic sensor taking advantage of the periodic nanostructured array of commercial Blu-ray discs. Sensors with two gold film thicknesses (50 and 100nm) were fabricated and optically characterized by varying the oblique-angle of the incident light in optical reflectance measurements. Contrary to the use normal light incidence previously reported with other optical discs, we observed an enhancement in sensitivity and a narrowing of the resonant linewidths as the light incidence angle was increased, which could be related to the generation of Fano resonant modes. The new sensors achieve a figure of merit (FOM) up to 35 RIU−1 and a competitive bulk limit of detection (LOD) of 6.3×10−6 RIU. These values significantly improve previously reported results obtained with normal light incidence reflectance measurements using similar structures. The sensor has been combined with versatile, simple, ease to-fabricate microfluidics. The integrated chip is only 1cm2 (including a PDMS flow cell with a 50µm height microfluidic channel fabricated with double-sided adhesive tape) and all the optical components are mounted on a 10cm×10cm portable prototype, illustrating its facile miniaturization, integration and potential portability. Finally, to assess the label-free biosensing capability of the new sensor, we have evaluated the presence of specific antibodies against the GTF2b protein, a tumor-associate antigen (TAA) related to colorectal cancer. We have achieved a LOD in the pM order and have assessed the feasibility of directly measuring biological samples such as human serum.

Graphene-Based Biosensors: Going Simple.

The main properties of graphene derivatives facilitating optical and electrical biosensing platforms are discussed, along with how the integration of graphene derivatives, plastic, and paper can lead to innovative devices in order to simplify biosensing technology and manufacture easy-to-use, yet powerful electrical or optical biosensors. Some crucial issues to be overcome in order to bring graphene-based biosensors to the market are also underscored.

ZnO films formed by atomic layer deposition as an optical biosensor platform for the detection of Grapevine virus A-type proteins

Novel sensitive optical biosensor for determination of Grapevine virus A-type (GVA) proteins (GVA-antigens) has been designed. This biosensor was based on thin films of Zinc Oxide (ZnO) deposited by atomic layer deposition (ALD). The ZnO-based films have demonstrated favorable surface-structural properties for the direct immobilization of antibodies against GVA-antigens in order to form a biosensitive layer sensitive to GVA-antigens. The immobilization was confirmed by intensity changes in the main near band emission (NBE) peak of ZnO and by the formation of intense photoluminescence band, discovered in the visible range around 425nm, caused by the immobilized proteins. The GVA-antigen detection was performed by the evaluation of changes and behavior of a corresponding luminescence band. The sensitivity of as-formed label-free biosensor towards the GVA-antigens was determined in the range from 1pg/ml to 10ng/ml; in addition, the selectivity of biosensor was evaluated.

Ambient light-based optical biosensing platform with smartphone-embedded illumination sensor

We present a hand-held optical biosensing system utilizing a smartphone-embedded illumination sensor that is integrated with immunoblotting assay method. The smartphone-embedded illumination sensor is regarded as an alternative optical receiver that can replaces the conventional optical analysis apparatus because the illumination sensor can respond to the ambient light in a wide range of wavelengths, including visible and infrared. To demonstrate the biosensing applicability of our system employing the enzyme-mediated immunoblotting and accompanying light interference, various types of ambient light conditions including outdoor sunlight and indoor fluorescent were tested. For the immunoblotting assay, the biosensing channel generating insoluble precipitates as an end product of the enzymatic reaction is fabricated and mounted on the illumination sensor of the smartphone. The intensity of penetrating light arrives on the illumination sensor is inversely proportional to the amount of precipitates produced in the channel, and these changes are immediately analyzed and quantified via smartphone software. In this study, urinary C-terminal telopeptide fragment of type II collagen (uCTX-II), a biomarker of osteoarthritis diagnosis, was tested as a model analyte. The developed smartphone-based sensing system efficiently measured uCTX-II in the 0–5ng/mL concentration range with a high sensitivity and accuracy under various light conditions. These assay results show that the illumination sensor-based optical biosensor is suitable for point-of-care testing (POCT).

Recent advances in nanoplasmonic biosensors: applications and lab-on-a-chip integration

Abstract Motivated by the recent progress in the nanofabrication field and the increasing demand for cost-effective, portable, and easy-to-use point-of-care platforms, localized surface plasmon resonance (LSPR) biosensors have been subjected to a great scientific interest in the last few years. The progress observed in the research of this nanoplasmonic technology is remarkable not only from a nanostructure fabrication point of view but also in the complete development and integration of operative devices and their application. The potential benefits that LSPR biosensors can offer, such as sensor miniaturization, multiplexing opportunities, and enhanced performances, have quickly positioned them as an interesting candidate in the design of lab-on-a-chip (LOC) optical biosensor platforms. This review covers specifically the most significant achievements that occurred in recent years towards the integration of this technology in compact devices, with views of obtaining LOC devices. We also discuss the most relevant examples of the use of the nanoplasmonic biosensors for real bioanalytical and clinical applications from assay development and validation to the identification of the implications, requirements, and challenges to be surpassed to achieve fully operative devices.

Diffraction microgratings as a novel optical biosensing platform

Using a micro-hole grating in a supported silver film as a laser-fabricated novel optical platform for surface-enhanced IR absoprtion/reflection spectroscopy, characteristic absorption bands of Staphylococcus aureus, in particular, its buried carotenoid fragments, were detected in FT-IR spectra with 10-fold analytical enhancement, paving the way for the spectral express-identification of pathogenic microorganisms.

Reusable split-aptamer-based biosensor for rapid detection of cocaine in serum by using an all-fiber evanescent wave optical biosensing platform

A rapid, facile, and sensitive assay of cocaine in biological fluids is important to prevent illegal abuse of drugs. A two-step structure-switching aptasensor has been developed for cocaine detection based on evanescent wave optical biosensing platform. In the proposed biosensing platform, two tailored aptamer probes were used to construct the molecular structure switching. In the existence of cocaine, two fragments of cocaine aptamer formed a three-way junction quickly, and the fluorophore group of one fragment was effectively quenched by the quencher group of the other one. The tail of the three-way junction hybridized with the cDNA sequences immobilized on the optical fiber biosensor. Fluorescence was excited by evanescent wave, and the fluorescence signal was proportional to cocaine concentration. Cocaine was detected in 450 s (300 s for incubation and 150 s for detection and regeneration) with a limit of detection (LOD) of 165.2 nM. The proposed aptasensor was evaluated in human serum samples, and it exhibited good recovery, precision, and accuracy without complicated sample pretreatments.

Optical biosensing platforms based on Ga-graphene plasmonic structures on Cu, quartz and SiO2/Si substrates

Ga nanoparticles (GaNPs) produced by thermal evaporation or epitaxial methods on graphene sheets present strong plasmonic effects that can couple to graphene phonons. In this work, GaNPs are deposited on graphene monolayers supported on Cu, quartz, and SiO2/Si substrates. The use of graphene increases the interparticle distance and improves the size uniformity. The enhancement of G and 2D graphene Raman lines is analyzed for the different substrates under excitation at 633 and 532 nm. The plasmonic effects are investigated using spectroscopic ellipsometry, finding a strong resonant mode at energies between 1.7 and 2.0 eV and an incidence angle of 55.6°. Finally, the heterogeneous surfaces are functionalized for the preparation of optical biosensors exposing the surface to 3,3′-dithiodipropionic acid di(N-succinimidyl ester) (DTSP). A significant energy shift of the ellipsometric signal on the Ga/graphene surfaces is observed in comparison to Ga/Si surfaces. Sketch of GaNPs atop graphene and biofunctionalizing agent (DTSP). Dip in the Psi ellipsometric function, and associated step in the Delta function, found around the GaNPs–graphene coupling resonance at an incident angle of 55.6°.

Development of L-lactate dehydrogenase biosensor based on porous silicon resonant microcavities as fluorescence enhancers.

The up-regulation of L-lactate dehydrogenase (LDH), an intracellular enzyme present in most of all body tissues, is indicative of several pathological conditions and cellular death. Herein, we demonstrate LDH detection using porous silicon (pSi) microcavities as a luminescence-enhancing optical biosensing platform. Non-fluorescent resazurin was covalently attached onto the pSi surface via thermal hydrocarbonisation, thermal hydrosylilation and acylation. Each surface modification step was confirmed by means of FTIR and the optical shifts of the resonance wavelength of the microcavity. Thermal hydrocarbonisation also afforded excellent surface stability, ensuring that the resazurin was not reduced on the pSi surface. Using a pSi microcavity biosensor, the fluorescence signal upon detection of LDH was amplified by 10 and 5-fold compared to that of a single layer and a detuned microcavity, respectively, giving a limit of detection of 0.08 U/ml. The biosensor showed a linear response between 0.16 and 6.5 U/ml, covering the concentration range of LDH in normal as well as damaged tissues. The biosensor was selective for LDH and did not produce a signal upon incubation with another NAD-dependant enzyme L-glutamic dehydrogenase. The use of the pSi microcavity as a sensing platform reduced reagent usage by 30% and analysis time threefold compared to the standard LDH assay in solution.

Label-free optical biosensors based on aptamer-functionalized porous silicon scaffolds.

A proof-of-concept for a label-free and reagentless optical biosensing platform based on nanostructured porous silicon (PSi) and aptamers is presented in this work. Aptamers are oligonucleotides (single-stranded DNA or RNA) that can bind their targets with high affinity and specificity, making them excellent recognition elements for biosensor design. Here we describe the fabrication and characterization of aptamer-conjugated PSi biosensors, where a previously characterized his-tag binding aptamer (6H7) is used as model system. Exposure of the aptamer-functionalized PSi to the target proteins as well as to complex fluids (i.e., bacteria lysates containing target proteins) results in robust and well-defined changes in the PSi optical interference spectrum, ascribed to specific aptamer-protein binding events occurring within the nanoscale pores, monitored in real time. The biosensors show exceptional stability and can be easily regenerated by a short rinsing step for multiple biosensing analyses. This proof-of-concept study demonstrates the possibility of designing highly stable and specific label-free optical PSi biosensors, employing aptamers as capture probes, holding immense potential for application in detection of a broad range of targets, in a simple yet reliable manner.

A hot-spot-active magnetic graphene oxide substrate for microRNA detection based on cascaded chemiluminescence resonance energy transfer.

Herein, a cascaded chemiluminescence resonance energy transfer (C-CRET) process was demonstrated from horseradish peroxidase (HRP)-mimicking DNAzyme-catalyzed luminol-H2O2 to fluorescein and further to graphene oxide (GO) when HRP-mimicking DNAzyme/fluorescein was in close proximity to the GO surface. The proposed C-CRET system was successfully implemented to construct three modes of C-CRET hot-spot-active substrates (modes I, II and III) by covalently immobilizing HRP-mimicking DNAzyme/fluorescein-labeled hairpin DNAs (hot-spot-generation probes) on magnetic GO (MGO), resulting in a signal "off" state due to the quenching of the luminol/H2O2/HRP-mimicking DNAzyme/fluorescein CRET system by GO. Upon the introduction of microRNA-122 (miRNA-122), the targets (mode I) or the new triggers that were generated through a strand displacement reaction (SDR) initiated by miRNA-122 (modes II and III) hybridized with the loop domains of hairpin probes on MGO to form double-stranded (modes I and II) or triplex-stem structures (mode III), causing an "open" configuration of the hairpin probe and a CRET signal "on" state, thus achieving sensitive and selective detection of miRNA-122. More importantly, the substrate exhibited excellent controllability, reversibility and reproducibility through SDR and magnetic separation (modes II and III), especially sequence-independence for hairpin probes in mode III, holding great potential for the development of a versatile platform for optical biosensing.

Wine fingerprinting using a bio-geochemical approach

The wine sector is a billion euro business and therefore subjected to multiple attempts of fraudulent practices. This requires the development of rapid and reliable methods to detect such situations. Several methodologies have been developed based on the chemical profiles of the wines, but they are limited due to the environmental conditions that cannot be controlled. The use of DNA-based detection systems are an emergent research field that have been extended to a wide variety of food prod- ucts and are still the most reliable methods for varietal identification. However these methods are not suitable for geographical determination. Soil related fingerprints have a primary role considering that there is a relationship between the elemental compo- sition of wine and the composition of the provenance soil. WineBioCode is a project aiming to define the best strategy for wine authenticity based on a multidisciplinary approach. Two DNA-based strategies have been developed based on Real-time PCR and a label free optical biosensor platform. Both platforms enabled successful identification of specific DNA-targets when applied to Vitis vinifera L., and can be applied throughout the grape-wine chain. The methods are complementary and can be used in dif- ferent situations, according to the requirements. The geographical evaluation has been assessed by the strontium 87 Sr/86 Sr isotope ratio determination involving soil evaluation in the vineyards followed by its assay in the wine samples. The results are being integrated in order to establish the best procedure to be undertaken for wine fingerprinting, including varietal composition and geographical origin, therefore fulfilling the requirements of the geographical denominations in wine certification.

Optical waveguide sensor based on silica nanotube arrays for label-free biosensing.

Label-free biosensing based on optical waveguide spectroscopy of silica nanotube (SNT) arrays is realized with high sensitivity. The SNT arrays fabricated using a porous anodic alumina (PAA) template assisted by surface sol-gel (SSG) method showed a high value of 552 reciprocal refractive index unit as the sensing figure of merit by exchanging the sensing environment with water and ethanol. A standard biotin-streptavidin affinity model was tested using the SNT arrays which support a TM1 mode and the fundamental response of the system was investigated. Results show that the response of the SNT arrays for adsorption of streptavidin is higher than the one using substrate without removing the PAA template due to the larger surface area and the stronger electromagnetic field. The limit of detection (LOD) of the SNT arrays for detection of streptavidin was estimated as 93 pM, with the detection time of 40 min. Additionally, the Fresnel calculations suggested higher potential sensitivity of the current system compared to that of the conventional SPR sensors. Thus, the SNT arrays may be used as a versatile platform for high-sensitive label-free optical biosensing due to the high performance and the large potential of the surface functionality.

The transformation of common office supplies into a low-cost optical biosensing platform

By reassembling common office supplies, an optical biosensing system was developed. A laser pointer and the solar cell from a calculator were utilized in the developed optical biosensing system as the light source and signal transducer, respectively. For intuitive signal evaluation, a multimeter was used. The following two types of conventional enzymatic colorimetric assays were employed with the optical biosensing system: (i) the Trinder׳s reaction-based enzymatic assay; and (ii) the competitive enzyme-linked immunosorbent assay. These colorimetric assays were performed in reaction channels made from transparent polymer and glass. By matching the maximum absorption spectra of the colored end products from the assays with the emission spectra of the laser diodes, the biochemical reaction rate was manifested as a change in the intensity of the laser beam. This change was then converted by the solar cell into voltage and displayed on the connected multimeter. To verify the detection performance of the system, glucose and an osteoarthritis biomarker (urinary collagen type II C-telopeptide fragments [uCTX-II]) were quantified. With glucose, the voltages registered were linearly correlated with the glucose concentration, from 0 to 10mM. Using a competitive immunoassay for uCTX-II, the system exhibited a calibration curve with a dynamic detection range between 1.3 and 10ng/mL uCTX-II. Given the advantages of the proposed biosensing system, including its high sensitivity, facile fabrication, and the high obtainability and cost-effectiveness of the components used to make it, we expect that this study will provide a basis for the production of a low-cost optical biosensor.

Label-free optical biosensors based on a planar optical waveguide

We review optical label-free biosensing platforms based on planar optical waveguides with their operation principles and performance characteristics. As the building blocks of refractometric optical transducers and plasmonic optical devices, optical planar waveguides are widely adopted to be core sensing platforms for label-free optical assay of biological and chemical interactions due to the high sensitivity with their invulnerability to external noise such as electromagnetic interference. The review scope includes the sensing schemes of conventional dielectric waveguides, the reverse symmetry waveguides, the anti-resonant reflecting optical waveguides (ARROW), and metal-clad leaky waveguides (MCLW). The configurations of the sensing systems with their operation features will be discussed together with the recent development and progress. In particular, we address in more detail the reverse symmetry waveguide and the MCLW based biosensors which have a capability of detecting micrometer-scale biological objects.

Development of a new parallelized, optical biosensor platform for label-free detection of autoimmunity-related antibodies

Autoimmune diseases are characterized by the presence of autoantibodies in serum of affected patients. The heterogeneity of autoimmune relevant antigens creates a variety of different antibodies, which requires a simultaneous detection mode. For this reason, we developed a tool for parallelized, label-free, optical detection that accomplishes the characterization of multiple antigen-antibody interactions within a single measurement on a timescale of minutes. Using 11-aminoundecyltrimethoxysilane, we were able to immobilize proteinogenic antigens as well as an amino-functionalized cardiolipin on a glass surface. Assay conditions were optimized for serum measurements with a single spot antigen chip on a single spot 1-λ detection system. Minimized background signal allows a differentiation between patients and healthy controls with a good sensitivity and specificity. Applying polarized imaging reflectometric interference spectroscopy, we evaluated samples from three APS patients and three control subjects for this proof-of-principle and already obtained good results for β2-glycoprotein I and cardiolipin.

Plastic optical fiber-based biosensor platform for rapid cell detection

This work presents a novel, fast response time, plastic optic fiber (POF) biosensor to detect Escherichia coli. It discloses the technique for the development, calibration and measurement of this robust and simple-to-construct POF biosensor. The probes in U-shaped format were manufactured with a specially developed device. The calibration process led to the evaluation of the sensitivity, accuracy and repeatability by using solutions of sucrose for obtaining refractive indices (RI) in the range 1.33–1.39 IR equivalent of water and bacteria, respectively. The POF probes were functionalized with antibody anti-E. coli serotype O55 and tested firstly with saline and then with bacterial concentrations of 104, 106, and 108 colony forming units/ml (CFU/ml). The optoelectronic setup consists of an 880nm LED connected to the U-shaped probe driven by a sine waveform generated by the Simulink (from Matlab®). On the other side of the probe a photodetector generates a photocurrent which is amplified by a transconductance amplifier. The output voltage signal is read by the analog-to-digital (A/D) input of the microcontroller. In all tested concentrations, the results presented a tendency of a decrease in the output signal with time, due to the attachment of the bacteria to the POF probe and consequent increase in the RI close to the sensitive area of the fiber surface. It has been shown that the system is capable of providing positive response to the bacterial concentration in less than 10min, demonstrating good possibilities to be commercially developed as a portable field sensor.