Optical Biosensing Platform Research Articles

A Bioengineered Stable Protein 1‐Hemin Complex with Enhanced Peroxidase‐Like Catalytic Properties

Enzymes have gained their unique efficiency and catalytic activity through billions of years of evolution, perfecting their active site to a desired reaction. Inspired by nature, a novel enzyme‐mimicking platform is designed based on stable protein 1 (SP1) to create a nano‐compartment that mimics peroxidase activity. The biohybrid reveals enhanced activity over the hemin cofactor alone and improved stability in organic solvents in comparison to native peroxidase. Furthermore, the utilization of the obtained biohybrid in an optical glucose biosensing platform is shown. The biohybrid crystallographic structure is solved, indicating that the SP1 structure is not affected by the hemin coordination. This work opens the path for developing new cofactor binding centers in engineered protein scaffolds for various artificial catalytic processes.

(Invited) Pathogen Agnostic Biodetection at the Point of Need

Emerging pathogens are a constant threat to global health and human life, as exemplified by the recent COVID-19 pandemic. The availability of rapid testing - including in home tests - was instrumental in facilitating return to our 'normal' way of life. Diagnostics - especially those that can quickly and effectively inform on the condition at the point of need, enhance situational awareness and guide decision making and mitigation strategies can greatly impact our ability to counter emerging threats, epidemics and pandemics. Yet, current diagnostics are largely targeted- meaning they are tailored to identify a limited repertoire of anticipated threats. For true preparedness, it is important that the strategies we develop are pathogen agnostic and can be readily used against any organism quickly. Our team has sought inspiration from the human innate immune system in order to develop truly agnostic diagnostics. Innate immunity is medicated by pattern recognition receptors that have evolved to recognize evolutionarily conserved signatures on pathogens - allowing for their early identification. These germline receptors bind with conserved pathogen associated molecular patterns – PAMPs – produced by all pathogens, and result in the development of an associated cytokine and chemokine response. Innate immunity is thus a universal biosensor. Drawing inspiration from this natural system, our team has developed diagnostic assays for all pathogens. This involved understanding of common mechanisms of host-pathogen interactions, which then lead to the design of curated novel assay modalities - membrane insertion and lipoprotein capture - for the detection of pathogen. Beyond assay development and characterization of the response, we have evaluated the feasibility of these methods to detect signatures in blinded clinical samples for a variety of diseases (tuberculosis, invasive Salmonella, Shiga toxin carrying E. coli, Staphylococcal bacteremia, and others) with excellent sensitivity as compared to benchmark assays. These outcomes and strategies for pathogen agnostic detection will be presented.Transitioning this platform for field use, our team engineered a portable waveguide-based optical biosensor platform with integrated microfluidics for sample processes. The platform named Portable EnGineered Analytical Sensor with aUtomated Sampling (PEGASUS, R&D100 2021). This platform uses single mode planar optical waveguides as the sensing element, and builds on a bench top instrument that has been developed at the Los Alamos National Laboratory. PEGASUS miniaturizes this benchtop sensor, with integrated sample processing, moving us toward the goal of developing a truly fieldable biosensor. PEGASUS uses a different, smaller waveguide mounting apparatus, miniaturized robust components, sensing hardware and software.PEGASUS integrates Catch-all, the first ever sample processing system capable of lipid separation with ultra-sensitive optical detection of complex bioanalytes at the point of need. a microfluidic device capable of centrifugal separation of serum from blood at the point of need with a system that is compatible with biomarkers that are both hydrophilic and hydrophobic. The cross-flow filtration device separates serum from blood as efficiently as traditional methods and retains amphiphilic biomarkers in serum for detection.Whereas the scope of targets is extremely broad, the platform still relies on the use of reagents, which complicates field use in some instances. Therefore, we are beginning to explore multi-wavelength spectroscopy as a tool for the measurement of complex clinical signatures - within seconds, without reagents. Preliminary data from this development will also be presented.

Lab-on-a-chip optical biosensor platform: a micro-ring resonator integrated with a near-infrared Fourier transform spectrometer.

In this paper, we demonstrated the design and experimental results of the near-infrared lab-on-a-chip optical biosensor platform that monolithically integrates the MRR and the on-chip spectrometer on the silicon-on-insulator (SOI) wafer, which can eliminate the external optical spectrum analyzer for scanning the wavelength spectrum. The symmetric add-drop MRR biosensor is designed to have a free spectral range (FSR) of ∼19 nm and a bulk sensitivity of ∼73 nm/RIU; then the drop-port output resonance peaks are reconstructed from the integrated spatial-heterodyne Fourier transform spectrometer (SHFTS) with the spectral resolution of ∼3.1 nm and the bandwidth of ∼50 nm, which results in the limit of detection of 0.042 RIU.

Molybdenum Disulfide as Tunable Electrochemical and Optical Biosensing Platforms for Cancer Biomarker Detection: A Review.

Cancer is a common illness with a high mortality. Compared with traditional technologies, biomarker detection, with its low cost and simple operation, has a higher sensitivity and faster speed in the early screening and prognosis of cancer. Therefore, extensive research has focused on the development of biosensors and the construction of sensing interfaces. Molybdenum disulfide (MoS2) is a promising two-dimensional (2D) nanomaterial, whose unique adjustable bandgap shows excellent electronic and optical properties in the construction of biosensor interfaces. It not only has the advantages of a high catalytic activity and low manufacturing costs, but it can also further expand the application of hybrid structures through different functionalization, and it is widely used in various biosensors fields. Herein, we provide a detailed introduction to the structure and synthesis methods of MoS2, and explore the unique properties and advantages/disadvantages exhibited by different structures. Specifically, we focus on the excellent properties and application performance of MoS2 and its composite structures, and discuss the widespread application of MoS2 in cancer biomarkers detection from both electrochemical and optical dimensions. Additionally, with the cross development of emerging technologies, we have also expanded the application of other emerging sensors based on MoS2 for early cancer diagnosis. Finally, we summarized the challenges and prospects of MoS2 in the synthesis, functionalization of composite groups, and applications, and provided some insights into the potential applications of these emerging nanomaterials in a wider range of fields.

PCL/PEO Polymer Membrane Prevents Biofouling in Wearable Detection Sensors.

Technological advances in biosensing offer extraordinary opportunities to transfer technologies from a laboratory setting to clinical point-of-care applications. Recent developments in the field have focused on electrochemical and optical biosensing platforms. Unfortunately, these platforms offer relatively poor sensitivity for most of the clinically relevant targets that can be measured on the skin. In addition, the non-specific adsorption of biomolecules (biofouling) has proven to be a limiting factor compromising the longevity and performance of these detection systems. Research from our laboratory seeks to capitalize on analyte selective properties of biomaterials to achieve enhanced analyte adsorption, enrichment, and detection. Our goal is to develop a functional membrane integrated into a microfluidic sampling interface and an electrochemical sensing unit. The membrane was manufactured from a blend of Polycaprolactone (PCL) and Polyethylene oxide (PEO) through a solvent casting evaporation method. A microfluidic flow cell was developed with a micropore array that allows liquid to exit from all pores simultaneously, thereby imitating human perspiration. The electrochemical sensing unit consisted of planar gold electrodes for the monitoring of nonspecific adsorption of proteins utilizing Cyclic Voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). The solvent casting evaporation technique proved to be an effective method to produce membranes with the desired physical properties (surface properties and wettability profile) and a highly porous and interconnected structure. Permeability data from the membrane sandwiched in the flow cell showed excellent permeation and media transfer efficiency with uniform pore activation for both active and passive sweat rates. Biofouling experiments exhibited a decrease in the extent of biofouling of electrodes protected with the PCL/PEO membrane, corroborating the capacity of our material to mitigate the effects of biofouling.

Machine learning assisted cancer cell detection using strip waveguide Bragg gratings

An early cancer diagnosis has become necessary in the medical domain as it can facilitate a timely and appropriate treatment, providing for the clinical management of patients. This work proposes a rapid and inexpensive label-free bio-sensing platform based on Waveguide Bragg Gratings for cancer detection. The refractive index values of cancer cells are different from normal cells, which provide a different reflection spectrum when the sample is placed on the functionalized surface of the waveguide grating structure. We combine simulations from the effective index method, finite element method, and Transfer Matrix method of the Grating structure to optimize the sensing structure and employ a neural network model to predict whether a sample contains cancerous cells. The machine learning model provides excellent accuracy for high-resolution interrogator data, and the accuracy falls below 95% only when the resolution of the interrogator is taken as 0.9 nm, thereby eliminating the need for an expensive high-resolution optical Interrogator system. This, combined with the mass-scale production capability of the waveguide Bragg gratings using standard CMOS technology, paves the way towards combining nano-scale optical biosensing platforms and machine learning-based optimization techniques for early and low-cost label-free cancer detections.

Guanine Quantum Defects in Carbon Nanotubes for Biosensing.

Fluorescent single-wall carbon nanotubes (SWCNTs) are used as nanoscale biosensors in diverse applications. Selectivity is built in by noncovalent functionalization with polymers such as DNA. Recently, covalent functionalization was demonstrated by conjugating guanine bases of adsorbed DNA to the SWCNT surface as guanine quantum defects (g-defects). Here, we create g-defects in (GT)10-coated SWCNTs (Gd-SWCNTs) and explore how this affects molecular sensing. We vary the defect densities, which shifts the E11 fluorescence emission by 55 nm to a λmax of 1049 nm. Furthermore, the Stokes shift between absorption and emission maximum linearly increases with defect density by up to 27 nm. Gd-SWCNTs represent sensitive sensors and increase their fluorescence by >70% in response to the important neurotransmitter dopamine and decrease it by 93% in response to riboflavin. Additionally, the extent of cellular uptake of Gd-SWCNTs decreases. These results show how physiochemical properties change with g-defects and that Gd-SWCNTs constitute a versatile optical biosensor platform.

Suppressing the mechanochromism of flexible photonic crystals.

Photonic crystal slabs (PCS) are a promising platform for optical biosensing. Yet, flexible applications based on PCS for biosensing have been limited, as the mechanical properties influence the optical ones. Here, we show the suppression of the mechanochromism effect for flexible PCS. We obtained flexible photonic crystal slabs by sputtering of a dielectric 100 nm Nb2O5 high refractive index layer onto a flexible nanostructured polydimethylsiloxane (PDMS) substrate with 370 nm grating period. The PCS exhibit a guided mode resonance at around 650 nm. We demonstrate that these flexible photonic crystal slabs show less than 0.5 nm resonance shift for 4% strain and call them stabilized PCS (sPCS). We compare this to a resonance shift of ∼21 nm for ∼4% strain of a flexible photonic crystal with a flexible nanoparticle high index layer (mechanochromatic PCS, mPCS). This high resonance shift is expected from the Bragg equations, where 4% grating period change correspond to approximately 4% change of the resonance wavelength (i.e., ∼26 nm at a resonance wavelength of 650 nm), if changes in the mode effective refractive index are neglected. In a stretch series we obtain color-to-strain dependencies of 4.79 nm/% strain for mPCS and 0.11 nm/% strain for our stabilized sPCS. We analyze the suppression of the mechanochromism with detailed microscopy results. We observe that fissures and fractures form in the rigid waveguiding layer of the sPCS upon mechanical stress. An algorithm based on Holistically-Nested Edge Detection (HED) is used for automated counting of cracks. Rigid photonic crystal cells with sizes on the order of 10 µm to 100 µm are formed that explain the stable optical properties. Even more stable optical properties with less than 0.03 nm wavelength shift per 1% strain are demonstrated for sPCS with an additional dielectric 100 nm SiO2 low index layer beneath the Nb2O5 waveguide layer decoupling the waveguide further from the flexible PDMS substrate.

High-Figure-of-Merit Biosensing and Enhanced Excitonic Absorption in an MoS2-Integrated Dielectric Metasurface.

Among the transitional metal dichalcogenides (TMDCs), molybdenum disulfide (MoS2) is considered an outstanding candidate for biosensing applications due to its high absorptivity and amenability to ionic current measurements. Dielectric metasurfaces have also emerged as a powerful platform for novel optical biosensing due to their low optical losses and strong near-field enhancements. Once functionalized with TMDCs, dielectric metasurfaces can also provide strong photon-exciton interactions. Here, we theoretically integrated a single layer of MoS2 into a CMOS-compatible asymmetric dielectric metasurface composed of TiO2 meta-atoms with a broken in-plane inversion symmetry on an SiO2 substrate. We numerically show that the designed MoS2-integrated metasurface can function as a high-figure-of-merit (FoM=137.5 RIU-1) van der Waals-based biosensor due to the support of quasi-bound states in the continuum. Moreover, owing to the critical coupling of the magnetic dipole resonances of the metasurface and the A exciton of the single layer of MoS2, one can achieve a 55% enhanced excitonic absorption by this two-port system. Therefore, the proposed design can function as an effective biosensor and is also practical for enhanced excitonic absorption and emission applications.

Label-free optical biosensing: going beyond the limits.

Label-free optical biosensing holds great promise for a variety of applications in biomedical diagnostics, environmental and food safety, and security. It is already used as a key tool in the investigation of biomolecular binding events and reaction constants in real time and offers further potential additional functionalities and low-cost designs. However, the sensitivity of this technology does not match the routinely used but expensive and slow labelling methods. Therefore, label-free optical biosensing remains predominantly a research tool. Here we discuss how one can go beyond the limits of detection provided by standard optical biosensing platforms and achieve a sensitivity of label-free biosensing that is superior to labelling methods. To this end we review newly emerging optical implementations that overcome current sensitivity barriers by employing novel structural architectures, artificial materials (metamaterials and hetero-metastructures) and using phase of light as a sensing parameter. Furthermore, we elucidate the mechanism of plasmonic phase biosensing and review hyper-sensitive transducers, which can achieve detection limits at the single molecule level (less than 1 fg mm-2) and make it possible to detect analytes at several orders of magnitude lower concentrations than so far reported in literature. We finally discuss newly emerging layouts based on dielectric nanomaterials, bound states in continuum, and exceptional points.

In-situ label-free optical biosensing with plasmonic enhanced ellipsometry using partially-embedded bimetallic Ag-Au alloy nanoparticles

This work presents a label-free optical biosensing platform by integrating the localized surface plasmon resonance (LSPR) and in-situ ellipsometry technique. A sensor chip composed of partially embedded Au-Ag alloy nanoparticles was fabricated on a glass substrate by the thermal dewetting method. The sensor chip was characterized using spectroscopic ellipsometry to determine the optical constant. The bulk RI resolution estimated from the ellipsometry phase (∆) signal was found to be 4.62 × 10−7 RIU for the bare sensor chip. Also, the biosensing experiment toward protein detection gives a detection limit of 23 pM. Further, hexagonal-boron nitride nanosheets (h-BNNS) modified sensor chip shows enhanced sensitivity with the detection limit down to 4.2 pM. The protein adsorption coefficients obtained from the fitting of kinetics data give values of 4.6 × 104 mol−1s−1 and 2.65 × 105 mol−1 s−1 for bare, and h-BNNS modified sensor chips, respectively. A theoretical simulation performed with COMSOL shows good agreement with our experimental outcomes. We believe that our proposed sensor chip enables the features of high stability, surface-sensitive, non-destructive, label-free, easy to fabricate, and cost-effectiveness, which make it feasible for large-scale production and commercialization in biomedical applications.

Future Prospects of Luminescent Silicon Nanowires Biosensors.

In this paper, we exploit the perspective of luminescent Si nanowires (NWs) in the growing field of commercial biosensing nanodevices for the selective recognition of proteins and pathogen genomes. We fabricated quantum confined fractal arrays of Si NWs with room temperature emission at 700 nm obtained by thin-film, metal-assisted, chemical etching with high production output at low cost. The fascinating optical features arising from multiple scattering and weak localization of light promote the use of Si NWs as optical biosensing platforms with high sensitivity and selectivity. In this work, label-free Si NW optical sensors are surface modified for the selective detection of C-reactive protein through antigen-gene interaction. In this case, we report the lowest limit of detection (LOD) of 1.6 fM, fostering the flexibility of different dynamic ranges for detection either in saliva or for serum analyses. By varying the NW surface functionalization with the specific antigen, the luminescence quenching of NW biosensors is used to measure the hepatitis B-virus pathogen genome without PCR-amplification, with an LOD of about 20 copies in real samples or blood matrix. The promising results show that NW optical biosensors can detect and isolate extracellular vesicles (EV) marked with CD81 protein with unprecedented sensitivity (LOD 2 × 105 sEV/mL), thus enabling their measurement even in a small amount of blastocoel fluid.

Smartphone-based mobile biosensors for the point-of-care testing of human metabolites.

Rapid, accurate, portable and quantitative profiling of metabolic biomarkers is of great importance for disease diagnosis and prognosis. The recent development in the optical and electric biosensors based on the smartphone is promising for profiling of metabolites with advantages of rapid, reliability, accuracy, low-cost and multi-analytes analysis capability. In this review, we introduced the optical biosensing platforms including colorimetric, fluorescent and chemiluminescent sensing, and electrochemical biosensing platforms including wired and wireless communication. Challenges and future perspectives desired for reliable, accurate, cost-effective, and multi-functions smartphone-based biosensing systems were also discussed. We envision that such smartphone-based biosensing platforms will allow daily and comprehensive metabolites monitoring in the future, thus unlocking the potential to transform clinical diagnostics into non-clinical self-testing. We also believed that this progress report will encourage future research to develop advanced, integrated and multi-functional smartphone-based Point-of-Care testing (POCT) biosensors for the monitoring and diagnosis as well as personalized treatments of a spectrum of metabolic-disorder related diseases.

Mesoporous silica@chitosan@gold nanoparticles as “on/off” optical biosensor and pH-sensitive theranostic platform against cancer

A cancer nanotheranostic system was fabricated based on mesoporous silica@chitosan@gold (MCM@CS@Au) nanosystem targeted by aptamer toward the MUC-1 positive tumor cells. Subsequently, curcumin as an efficient herbal anticancer drug was first encapsulated into chitosan-triphosphate nanoparticles and then the resulted nanoparticle was loaded into the nanosystem (MCM@CS@Au-Apt). The nanosystem successful fabrication was approved at each synthesis step through FTIR, XRD, BET, DLS, FE-SEM, HRTEM, and fluorescence spectroscopy. Besides, the interaction between aptamer and curcumin was evaluated using full atomistic molecular dynamics simulations. The mechanism of curcumin release was likewise investigated through different kinetic models. Afterwards, the potential of the designed nanosystem in targeted imaging, and drug delivery was evaluated using fluorescence microscopy and flow cytometry. It was found that the energy transfer between the base pairs in the hairpin of double strands of DNA aptamer acts as a quencher for MCM@CS@Au fluorescence culminating in an “on/off” optical biosensor. On the other hand, the presence of pH-sensitive chitosan nanoparticles creates smart nanosystem to deliver more curcumin into the desired cells. Indeed, when the aptamer specifically binds to the MUC-1 receptor, its double strands separate under the low pH condition, leading to the drug release and the recovery of the fluorescence (“On” state). Based on the toxicity results, this nanosystem had more toxicity toward the MUC-1-positive tumor cells than MUC-1-negative cells, representing its selective targeting. Therefore, this nanosystem could be introduced as a smart anticancer nanotheranostic system for tracing particular biomarkers (MUC-1), non-invasive fluorescence imaging, and targeted curcumin delivery.

MXenes in photomedicine: advances and prospects.

MXenes and their related nanocomposites with superior physicochemical properties such as high surface area, ease of synthesis and functionalization, high drug loading capacity, collective therapy potentials, pH-triggered drug release behavior, high photothermal conversion, and excellent photodynamic efficiency have been explored as alluring materials in photomedicine; the application of photons in medicine is facilitated for imaging and various disease treatment methods such as photothermal cancer/tumor ablation. Non-invasive theranostic strategies with synergistic activities have been developed using photothermal, photodynamic, and magnetic therapies together with remotely controlled drug/gene delivery for the diagnosis and treatment of various malignant diseases. Photothermal/photodynamic therapy and photoacoustic imaging using MXene-based structures have shown great promise in cancer phototherapy. However, hybridization and surface functionalization should be further explored to obtain biocompatible MXene-based composites/platforms with unique properties, high stability, and improved functionality in photomedicine. Toxicological and long-term biosafety assessments as well as clinical translation evaluations ought to be given high priority in research. Although some limited studies have revealed the excellent potentials of MXenes and their derivatives in photomedicine, further steps should be taken towards extensive research and detailed analysis in the field of optimizing the properties and improving the performance of these materials with a clinical and industrial outlook. Optical biosensing platforms have been developed along with electrochemical sensors and wearable sensors constructed from MXenes and their derivatives; future studies warrant the comprehensive analysis of optical transduction aspects such as colorimetry, electrochemiluminescence, photoluminescence, surface-enhanced Raman scattering, and surface plasmon resonance. Herein, the potentials of MXenes in photomedicine are deliberated encompassing important challenges and future research directions.

Trends in point-of-care optical biosensors for antibiotics detection in aqueous media

Besides widespread use of the antibiotics for treatment of infectious diseases in both humans and animals, their unregulated disposal has become a major threat to the environment in several aspects like emergence of antibiotic resistance in microorganisms, hindered growth of healthy gut microbiome, and toxicity in humans and animals upon accumulation. Therefore, the need of development of selective and sensitive methods is highly desirable for rapid detection of antibiotics. Nowadays, optical biosensors have been emerged as relatively rapid, easy to use, and accurate approach as an alternative to conventional detection techniques. Here, the recent advancements in point-of-care (POC) optical biosensing platforms are highlighted for on-site detection of antibiotics in terms of affordable and user-friendly manner. The POC devices can be easily integrated with advanced systems such as intelligent or internet of things (IoT).

Deep Subwavelength Laser-Induced Periodic Surface Structures on Silicon as a Novel Multifunctional Biosensing Platform.

Strong light localization inside the nanoscale gaps provides remarkable opportunities for creation of various medical and biosensing platforms stimulating an active search for inexpensive and easily scalable fabrication at a sub-100 nm resolution. In this paper, self-organized laser-induced periodic surface structures (LIPSSs) with the shortest ever reported periodicity of 70 ± 10 nm were directly imprinted on the crystalline Si wafer upon its direct femtosecond-laser ablation in isopropanol. Appearance of such a nanoscale morphology was explained by the formation of a periodic topography on the surface of photoexcited Si driven by interference phenomena as well as subsequent down-scaling of the imprinted grating period via Rayleigh-Taylor hydrodynamic instability. The produced deep subwavelength LIPSSs demonstrate strong anisotropic anti-reflection performance, ensuring efficient delivery of the incident far-field radiation to the electromagnetic "hot spots" localized in the Si nanogaps. This allows realization of various optical biosensing platforms operating via strong interactions of quantum emitters with nanoscale light fields. The demonstrated 80-fold enhancement of spontaneous emission from the attached nanolayer of organic dye molecules and in situ optical tracing of catalytic molecular transformations substantiate bare and metal-capped deep subwavelength Si LIPSSs as a promising inexpensive multifunctional biosensing platform.

StaphAIR: A Label-Free Antigen Microarray Approach to Detecting Anti-Staphylococcus aureus Antibody Responses in Orthopedic Infections.

Arrayed imaging reflectometry (AIR) is an optical biosensor platform for simple, multiplex measurement of antigen-specific antibody responses in patient blood samples. Here, we report the development of StaphAIR, an 8-plex Staphylococcus aureus antigen array on the AIR platform for profiling antigen-specific anti-S. aureus humoral immune responses. Initial validation experiments with mouse and humanized monoclonal antibodies against the S. aureus autolysin glucosaminidase (Gmd) domain, and subsequent testing with dilution series of pooled positive human serum confirmed analytically robust behavior of the array, with all antigens displaying Langmuir-type dose-response curves. Testing a cohort of 82 patients with S. aureus musculoskeletal infections (MSKI) and 30 healthy individuals enabled discrimination of individual patient responses to different S. aureus antigens, with statistical significance between osteomyelitis patients and controls obtained overall for four individual antigens (IsdA, IsdB, Gmd, and SCIN). Multivariate analyses of the antibody titers obtained from StaphAIR revealed its utility as a potential diagnostic tool for detecting S. aureus MSKI (area under the receiver operating characteristic curve (AUC) > 0.85). We conclude that StaphAIR has utility as a high-throughput immunoassay for studying and diagnosing osteomyelitis in patients.

Optical Biosensor Platforms Display Varying Sensitivity for the Direct Detection of Influenza RNA.

Detection methods that do not require nucleic acid amplification are advantageous for viral diagnostics due to their rapid results. These platforms could provide information for both accurate diagnoses and pandemic surveillance. Influenza virus is prone to pandemic-inducing genetic mutations, so there is a need to apply these detection platforms to influenza diagnostics. Here, we analyzed the Fast Evaluation of Viral Emerging Risks (FEVER) pipeline on ultrasensitive detection platforms, including a waveguide-based optical biosensor and a flow cytometry bead-based assay. The pipeline was also evaluated in silico for sequence coverage in comparison to the U.S. Centers for Disease Control and Prevention’s (CDC) influenza A and B diagnostic assays. The influenza FEVER probe design had a higher tolerance for mismatched bases than the CDC’s probes, and the FEVER probes altogether had a higher detection rate for influenza isolate sequences from GenBank. When formatted for use as molecular beacons, the FEVER probes detected influenza RNA as low as 50 nM on the waveguide-based optical biosensor and 1 nM on the flow cytometer. In addition to molecular beacons, which have an inherently high background signal we also developed an exonuclease selection method that could detect 500 pM of RNA. The combination of high-coverage probes developed using the FEVER pipeline coupled with ultrasensitive optical biosensors is a promising approach for future influenza diagnostic and biosurveillance applications.

Ultrasensitive detection of endocrine disruptors via superfine plasmonic spectral combs

The apparent increase in hormone-induced cancers and disorders of the reproductive tract has led to a growing demand for new technologies capable of detecting endocrine disruptors. However, a long-lasting challenge unaddressed is how to achieve ultrahigh sensitive, continuous, and in situ measurement with a portable device for in-field and remote environmental monitoring. Here we demonstrate a simple-to-implement plasmonic optical fiber biosensing platform to achieve an improved light–matter interaction and advanced surface chemistry for ultrasensitive detection of endocrine disruptors. Our platform is based on a gold-coated highly tilted fiber Bragg grating that excites high-density narrow cladding mode spectral combs that overlap with the broad absorption of the surface plasmon for high accuracy interrogation, hence enabling the ultrasensitive monitoring of refractive index changes at the fiber surface. Through the use of estrogen receptors as the model, we design an estradiol–streptavidin conjugate with the assistance of molecular dynamics, converting the specific recognition of environmental estrogens (EEs) by estrogen receptor into surface-based affinity bioassay for protein. The ultrasensitive platform with conjugate-induced amplification biosensing approach enables the subsequent detection for EEs down to 1.5 × 10−3 ng ml−1 estradiol equivalent concentration level, which is one order lower than the defined maximal E2 level in drinking water set by the Japanese government. The capability to detect EEs down to nanogram per liter level is the lowest limit of detection for any estrogen receptor-based detection reported thus far. Its compact size, flexible shape, and remote operation capability open the way for detecting other endocrine disruptors with ultrahigh sensitivity and in various hard-to-reach spaces, thereby having the potential to revolutionize environment and health monitoring.