A Gold Nanoparticle-Based Cortisol Aptasensor for Non-Invasive Detection of Fish Stress.

Cortisol is a key stress biomarker in humans and animals, including fishes. In aquafarming, stress monitoring using cortisol quantification can help to optimize aquaculture practices for welfare and productivity enhancement. However, most current methods for cortisol detection rely on invasive tissue sampling. In this work, we developed a gold nanoparticle (AuNP)-based cortisol sensor to address the demand of detecting picomolar ranges of cortisol from complex fish tank water matrices as a non-invasive alternative for more effective stress monitoring. We first identified a DNA aptamer with effective binding to cortisol and then conjugated the thiol-labelled aptamer to AuNPs together with a blocker molecule (CALNN) to form an Au-Apt-CALNN conjugate that is stable in fish tank water. The cortisol detection principle is based on magnesium chloride (MgCl2)-induced particle aggregation, where the cortisol-bound aptamer on the AuNPs folds into a tertiary structure and provides greater protection for Au-Apt-CALNN against MgCl2-induced aggregation due to steric stabilization. At an optimum MgCl2 concentration, the differential stability of particles with and without cortisol binding offers a limit of detection (LOD) of 100 pM for cortisol within a 35 min reaction. The aptasensor has been validated on recirculating aquaculture system (RAS) fish tank water samples by the HPLC method and was able to detect changes in water cortisol induced by two different stress paradigms. This on-site deployable and non-invasive sensor offers opportunities for more efficient and real-time fish stress monitoring for the optimization of aquaculture practices.

A Rapid Total Bacterial Count Method for Food Samples using Syringe Filters and Lectin-Conjugated Semiconductor Nanorods.

Total bacterial count in food is one of important food safety criteria. The current plate count method (Heterotrophic Plate Count) for food analysis requires microbiology lab facilities and at least 2 days turnover time. We developed a rapid fluorescence-based total bacterial count method that utilises semiconductor nanorods (SNRs) conjugated with a lectin Griffonia simplicifolia II (GSII-SNRs) to stain bacterial cells captured on syringe filters, via the common N-acetylglucosamine molecules on bacterial cell wall. This "Filter-and-Stain" detection method has a rapid turnover time of 20 min. The fluorescence emission can be seen under UV light with minimum interference from food sample background. The fluorescence intensity quantified through image analysis is proportional to the bacterial concentration with a limit of detection of 1000 CFU/mL, for total bacterial count assessment in food safety. Moreover, the GSII-SNRs do not bind to heat inactivated bacterial cells, and thus can differentiate live and dead bacteria. Our method has been validated with representative food (coffee powder, raw spinach leaves, and ready-to-eat tomato salsa) to demonstrate its high potential for on-site food safety assessment, especially in places with no immediate access to microbiology labs.

Skin-Attachable Ink-Dispenser-Printed Paper Fluidic Sensor Patch for Colorimetric Sweat Analysis.

In situ analysis of sweat biomarkers potentially provides noninvasive lifestyle monitoring and early diagnosis. Quantitative detection of sweat rate is crucial for thermoregulation and preventing heat injuries. Here, a skin-attachable paper fluidic patch is reported for in situ colorimetric sensing of multiple sweat markers (pH, glucose, lactate, and uric acid) with concurrent sweat rate tracking. Two sets of fluidic patterns-multiplexed detection zones and a longitudinal sweat rate channel-are directly printed by an automated ink dispenser from a specially developed ceramic-based ink. The ceramic ink thermal-cures into an impervious barrier, confining sweat within the channels. The ceramic-ink-printed boundary achieves higher pattern resolution, prevents fluid leakage, attains pattern thermal stability, and resistant to organic solvents. The cellulose matrix of the detection zones is modified with nanoparticles to improve the color homogeneity and sweat sensor sensitivity. The sweat rate channel is made moisture sensitive by incorporating a metal-salt-based dye. The change in saturation/color of the detection zones and/or channels upon sweat addition can be visually detected or quantified by a smartphone camera. A cost-effective way is provided to fabricate paper fluidic sensor patches, successfully demonstrating on-body multiplexed evaluation of sweat analytes. Such skin wearables offer on-site analysis, meaningful to an increasingly health-conscious population.

A rapid total bacterial count method using gold nanoparticles conjugated with an aptamer for water quality assessment.

Total bacterial count is a routine parameter in microbial safety assessment used in many fields, such as drinking water and industrial water testing. The current gold standard method for counting bacteria is the plate culture method (or heterotrophic plate count) that requires a microbiology laboratory and a long turnover time of at least 24 hours. To tackle these shortcomings, we developed a rapid total bacterial count method that relies on gold nanoparticles (AuNPs) conjugated with affinity ligands to stain bacterial cells captured on a syringe filter. Two affinity ligands were exploited, i.e. a DNA aptamer (AB2) and a lectin Griffonia simplicifolia II (GSII) that recognize bacterial cell wall commonalities, i.e. peptidoglycan and its amino sugars. Upon proper formulation with addition of a surfactant, the AB2 conjugated AuNPs (AB2-AuNPs) can selectively stain bacterial cells captured on the filter membrane with a higher sensitivity than GSII-AuNPs. Measuring the staining intensity using an in-house-built handheld detector allowed us to correlate its intensity reading with the total number of bacterial units present. This bacteria quantification method, referred to as "Filter-and-Stain", had an efficient turnover time of 20 min suggesting its potential usage for rapid on-site applications. Additionally, the detection sensitivity provided by the AB2-AuNP nanoreagent offered a limit of detection as low as 100 CFU mL-1. We have demonstrated the use of the AB2-AuNPs for detection of bacteria from environmental water samples.

A Genetic Algorithm for Universal Optimization of Ultrasensitive Surface Plasmon Resonance Sensors with 2D Materials.

We present a general optimization technique for surface plasmon resonance, (SPR) yielding a range of ultrasensitive SPR sensors from a materials database with an enhancement of ∼100%. Applying the algorithm, we propose and demonstrate a novel dual-mode SPR structure coupling SPP and a waveguide mode within GeO2 featuring an anticrossing behavior and an unprecedented sensitivity of 1364 deg/RIU. An SPR sensor operating at wavelengths of 633 nm having a bimetal Al/Ag structure sandwiched between hBN can achieve a sensitivity of 578 deg/RIU. For a wavelength of 785 nm, we optimized a sensor as a Ag layer sandwiched between hBN/MoS2/hBN heterostructures achieving a sensitivity of 676 deg/RIU. Our work provides a guideline and general technique for the design and optimization of high sensitivity SPR sensors for various sensing applications in the future.

Battery-free and AI-enabled multiplexed sensor patches for wound monitoring.

Wound healing is a dynamic process with multiple phases. Rapid profiling and quantitative characterization of inflammation and infection remain challenging. We report a paper-like battery-free in situ AI-enabled multiplexed (PETAL) sensor for holistic wound assessment by leveraging deep learning algorithms. This sensor consists of a wax-printed paper panel with five colorimetric sensors for temperature, pH, trimethylamine, uric acid, and moisture. Sensor images captured by a mobile phone were analyzed by neural network-based machine learning algorithms to determine healing status. For ex situ detection via exudates collected from rat perturbed wounds and burn wounds, the PETAL sensor can classify healing versus nonhealing status with an accuracy as high as 97%. With the sensor patches attached on rat burn wound models, in situ monitoring of wound progression or severity is demonstrated. This PETAL sensor allows early warning of adverse events, which could trigger immediate clinical intervention to facilitate wound care management.

Understanding COVID-19: comparison of spatio-temporal analysis methods used to study epidemic spread patterns in the United States.

This article examines three spatiotemporal methods used for analyzing of infectious diseases, with a focus on COVID-19 in the United States. The methods considered include inverse distance weighting (IDW) interpolation, retrospective spatiotemporal scan statistics and Bayesian spatiotemporal models. The study covers a 12-month period from May 2020 to April 2021, including monthly data from 49 states or regions in the United States. The results show that the spread of COVID-19 pandemic increased rapidly to a high value in winter of 2020, followed by a brief decline that later reverted into another increase. Spatially, the COVID-19 epidemic in the United States exhibited a multi-centre, rapid spread character, with clustering areas represented by states such as New York, North Dakota, Texas and California. By demonstrating the applicability and limitations of different analytical tools in investigating the spatiotemporal dynamics of disease outbreaks, this study contributes to the broader field of epidemiology and helps improve strategies for responding to future major public health events.

Greatly Enhanced Resonant Exciton-Trion Conversion in Electrically Modulated Atomically Thin WS2 at Room Temperature.

Excitonic resonance in atomically thin semiconductors offers a favorite platform to study 2D nanophotonics in both classical and quantum regimes and promises potentials for highly tunable and ultra-compact optical devices. The understanding of charge density dependent exciton-trion conversion is the key for revealing the underlaying physics of optical tunability. Nevertheless, the insufficient and inefficient light-matter interactions hinder the observation of trionic phenomenon and the development of excitonic devices for dynamic power-efficient electro-optical applications. Here, by engaging an optical cavity with atomically thin transition metal dichalcogenides (TMDCs), greatly enhanced exciton-trion conversion is demonstrated at room temperature (RT) and achieve electrical modulation of reflectivity of ≈40% at exciton and 7% at trion state, which correspondingly enables a broadband large phase tuning in monolayer tungsten disulfide. Besides the absorptive conversion, ≈100% photoluminescence conversion from excitons to trions is observed at RT, illustrating a clear physical mechanism of an efficient exciton-trion conversion for extraordinary optical performance. The results indicate that both excitons and trions can play significant roles in electrical modulation of the optical parameters of TMDCs at RT. The work shows the real possibility for realizing electrical tunable and multi-functional ultra-thin optical devices using 2D materials.

Carbon Dot-Doped Hydrogel Sensor Array for Multiplexed Colorimetric Detection of Wound Healing.

Effective wound care and treatment require a quick and comprehensive assessment of healing status. Here, we develop a carbon dot-doped hydrogel sensor array in polydimethylsiloxane (PDMS) for simultaneous colorimetric detections of five wound biomarkers and/or wound condition indicators (pH, glucose, urea, uric acid, and total protein), leading to the holistic assessment of inflammation and infection. A biogenic carbon dot synthesized using an amino acid and a polymer precursor is doped in an agarose hydrogel matrix for constructing enzymatic sensors (glucose, urea, and uric acid) and dye-based sensors (pH and total protein). The encapsulated enzymes in such a matrix exhibit improved enzyme kinetics and stability compared to those in pure hydrogels. Such a matrix also provides stable colorimetric responses for all five sensors. The sensor array exhibits high accuracy (recovery rates of 91.5-113.1%) and clinically relevant detection ranges for all five wound markers. The sensor array is established for simulated wound fluids and validated with rat wound fluids from perturbed wound models. Distinct color patterns are obtained that can clearly distinguish healing vs nonhealing wounds visually and quantitatively. This hydrogel sensor array shows great potential for on-site wound sensing due to its long-term stability, lightweight, and flexibility.

Antibody conjugated Au/Ir@Cu/Zn-MOF probe for bacterial lateral flow immunoassay and precise synergistic antibacterial treatment.

Staphylococcus aureus is one of the most prevalent threats to public health. Rapid detection with high sensitivity and targeted killing is crucial to curb its spread. Herein, a metal-bearing nanocomposite, consisting of a bimetallic nanoparticle and a metal-organic framework (Au/Ir@Cu/Zn-MOF) was constructed. Upon conjugation with anti-S. aureus antibody, this nanocomposite (Ab-Au/Ir@Cu/Zn-MOF) was exploited for its dual functions, i.e. as a reporting probe in a lateral flow immunoassay and a high efficiency antibacterial reagent. Benefiting from the enrichment of Au/Ir NPs by the Cu/Zn-MOF, the Au/Ir@Cu/Zn-MOF-based lateral flow immunoassay sensor exhibited a visual limit of detection of 103 CFU/mL, which was100 times more sensitive than Au/Ir-based sensor. Moreover, the Ab-Au/Ir@Cu/Zn-MOF probe possessed synergistic photothermal-chemodynamic bactericidal effect that specifically targeted against S. aureus. Under a co-treatment by H2O2 (0.4 mM) and 808 nm near infrared irradiation (1 W/cm2, 5 min), complete sterilization of 5 × 105-106 CFU/mL S. aureus was achieved at a nanocomposite concentration as low as 6.25 µg/mL. The superior antibacterial efficiency was attributable to the three-fold properties of the Ab-Au/Ir@Cu/Zn-MOF probe: (1) enhanced multi-enzyme mimicking activities that promote reactive oxygen species generation, (2) high photothermal activity (efficiency of 53.70%), and (3) bacteria targeting ability via the antibody coating. By changing the antibody, this nanocomposite can be tailored to target a wide range of bacteria species, for detection and for precise antibacterial treatment.

Stimuli-Activable Metal-Bearing Nanomaterials and Precise On-Demand Antibacterial Strategies.

Bacterial infections remain the leading cause of death worldwide today. The emergence of antibiotic resistance has urged the development of alternative antibacterial technologies to complement or replace traditional antibiotic treatments. In this regard, metal nanomaterials have attracted great attention for their controllable antibacterial functions that are less prone to resistance. This review discusses a particular family of stimuli-activable metal-bearing nanomaterials (denoted as SAMNs) and the associated on-demand antibacterial strategies. The various SAMN-enabled antibacterial strategies stem from basic light and magnet activation, with the addition of bacterial microenvironment responsiveness and/or bacteria-targeting selectivity and therefore offer higher spatiotemporal controllability. The discussion focuses on nanomaterial design principles, antibacterial mechanisms, and antibacterial performance, as well as emerging applications that desire on-demand and selective activation (i.e., medical antibacterial treatments, surface anti-biofilm, water disinfection, and wearable antibacterial materials). The review concludes with the authors' perspectives on the challenges and future directions for developing industrial translatable next-generation antibacterial strategies.

Celebrating 25 Years of IMRE: Research Highlights on Nanomaterials and Nanotechnologies.

The Institute of Materials Research and Engineering (IMRE) is a research institute of the Science and Engineering Research Council (SERC), Agency for Science, Technology and Research (A*STAR). IMRE was established in September 1997. Over the past 25 years, IMRE has developed core competencies and interdisciplinary teams for material development from fundamental discoveries to industrial translation. Currently, with over 400 researchers and state-of-the-art research facilities, IMRE conducts world class research in important material and material technology fields, including polymer composites, optical materials, electronic materials, soft materials, structural materials, energy materials, biomaterials, quantum technologies, as well as advanced characterization. As a material-centered research institute in Singapore, IMRE has played important roles in pushing science boundaries and developing cutting-edge technologies. One of the key strategies is to partner international organizations, research institutes, and industry to fulfill its vision to be a leading research institute to accelerate materials research, moving from "Made in Singapore" toward "Created in Singapore".

Detecting bacterial infections in wounds: a review of biosensors and wearable sensors in comparison with conventional laboratory methods.

Bacterial infection is a common impediment towards wound healing. Detecting bacterial infections is important to promote wound healing and curb chronic non-healing wounds. In this review, we firstly discuss bacterial communities, including aerobic and anaerobic bacteria in various types of wounds. Following the discussion of wound sampling methods (swab, biopsy) for different wounds, we then discuss laboratory based conventional methods (bacteria cultures, Gram staining, analytical profile index systems, polymerase chain reaction, and gas chromatography coupled with mass spectrometry), focusing on their recent improvement. After that we discussed the contemporary biosensor methods, including e-Nose, electrochemical sensors, surface enhanced Raman spectroscopy, and nucleic acid lateral flow immunoassay. Biosensors embedded into wound dressing, termed wearable sensors or smart wound dressing, are also discussed for their ability of enabling bacteria detection directly from wound sites without the need for obtaining swab/biopsy samples. We have compared all the detection methods for their performance according to their respective targets (either bacteria cells or volatile/non-volatile metabolites); after that we evaluate the suitability of various methods in providing timely and accurate diagnostic results towards real-time, point-of-care testing of bacterial infections.

Additive-Free, Visible-Light-Enabled Decarboxylative Alkylation of Enamides.

Enamides are versatile precursors for synthesizing bioactive compounds. As their alkylations often require perstoichiometric amounts of oxidants, transition metals, or photocatalysts, we herein report a simple alternative for their alkylations by just using visible light to irradiate the mixture of the readily available N-hydroxyphthalimide esters and enamides without an additive. The reaction involves the photoactivation of a π-π stacking EDA complex between the substrates.

A portable SERS sensor for pyocyanin detection in simulated wound fluid and through swab sampling.

A portable surface-enhanced Raman spectroscopy (SERS) sensor for detecting pyocyanin (PYO) in simulated wound fluid and from bacteria samples was developed. Solution-phase SERS detection protocols are designed to be compatible with two different clinical practices for wound exudate collection, namely negative pressure liquid collection and swabbing. For citrate-coated metal nanoparticles of three different compositions, i.e. gold (AuNPs), alloyed silver/gold (AgAuNPs), and silver (AgNPs), we firstly confirmed their interaction with PYO in the complex wound fluid, using fluorescence quenching experiments, which rationalized the Raman enhancement effects. We then demonstrated the Raman enhancement effects of the metal nanoparticles in the order of AgNPs > AgAuNPs > AuNPs. The limit of detection (LOD) achieved for PYO is 1.1 µM (in a linear range of 0.1-25 µM by the AgNPs), 10.9 µM (in a linear range of 5-100 µM, by the AgAuNPs), and 17.7 µM (in a linear range of 10-100 µM by the AuNPs). The AgNP and AgAuNP sensors together cover the sensitivity and dynamic range requirements for the clinical detection of wound infection, where PYO is present at a concentration of 1-50 µM. In addition, sterilized cotton swabs were used to collect wound fluid and transfer samples into AgNP solution for SERS measurements. This detection protocol was completed within 5 minutes with a LOD of 23.1 µM (in a linear range of 15-100 µM). The SERS sensing protocol was validated by its successful detection of PYO in cultured Pseudomonas aeruginosa bacteria. The findings presented in this work pave the way towards point-of-care diagnostics of wound infections.

Visible-Light-Induced Selective Photolysis of Phosphonium Iodide Salts for Monofluoromethylations.

The sigma (σ)-hole effect has emerged as a promising tool to construct novel architectures endowed with new properties. A simple yet effective strategy for the generation of monofluoromethyl radicals is a continuing challenge within the synthetic community. Fluoromethylphosphonium salts are easily available, air- and thermally stable, as well as simple-to-handle. Herein, we report the ability of the σ-hole effect to facilitate the visible-light-triggered photolysis of phosphonium iodide salts, a charge-transfer complex, selectively giving fluoromethyl radicals. The usefulness and versatility of this new protocol are demonstrated through the mono-, di-, and trifluoromethylation of a variety of alkenes.

Gold Nanoparticle-based "Mix and Measure" Fluorimetric Assays to Quantify Antibody Titer.

Monoclonal antibodies (mAbs) for treatment of human diseases are typically human or humanized Immunoglobulin G (IgG) produced in mammalian cell lines. A rapid, less tedious, and high throughput method to quantify mAbs is in demand to accelerate mAb production efficiency. To quantify mAb titer, we developed gold nanoparticle (AuNPs)-based "mix and measure" fluorimetric assays by exploiting AuNPs' fluorescence quenching ability. The AuNPs are functionalized by an Fc binding protein, i. e. protein G, which binds human IgG and fluorescently labeled rat IgG (Alexa Fluor 488-rat IgG) with differential affinity. The assays can be in competition or displacement format. The competitive binding of human IgG drug and the labelled rat IgG to protein G-coated AuNP lead to varied fluorescent intensity that is proportional to the amount of human IgG analte; or the displacement of the labelled rat IgG from protein G-coated AuNP by human IgG can lead to fluorescent recovery that is also proportionally related to human IgG concentration. The assays can quantify therapeutic mAbs in the range of 10-1,000 mg/L, demonstrated for Herceptin, Avastin, and Humira in cell culture media. The assays have fast turn over time (within 15 min). They can be performed in microplates and are suitable for high throughput "on-line" or "at-line" measurement in mAbs production lines.

Structure-selective differentiation of deletion mutations in circulating tumor DNA using dual probe-based isothermal amplification.

A rapid dual probe-based fluorimetric assay was developed to detect deletion mutations in circulating tumor DNA using structure-selective isothermal amplification and pattern recognition. This method could detect both homozygous and heterozygous deletion configurations in a one-set experiment and achieved picomolar detection limits with high selectivity within 2 hours. It was promising for point-of-care cancer diagnosis in hospital settings.

Epitope-Functionalized Gold Nanoparticles for Rapid and Selective Detection of SARS-CoV-2 IgG Antibodies.

Rapid and inexpensive immunodiagnostic assays to monitor severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seroconversion are essential for conducting large-scale COVID-19 epidemiological surveillance and profiling humoral responses against SARS-CoV-2 infections or immunizations. Herein, a colorimetic serological assay to detect SARS-CoV-2 IgGs in patients' plasma was developed using short antigenic epitopes conjugated to gold nanoparticles (AuNPs). Four immunodominant linear B-cell epitopes, located on the spike (S) and nucleocapsid (N) proteins of SARS-CoV-2, were characterized for their IgG binding affinity and used as highly specific biological motifs on the nanoparticle to recognize target antibodies. Specific bivalent binding between SARS-CoV-2 antibodies and epitope-functionalized AuNPs trigger nanoparticle aggregation, which manifests as a distinct optical transition in the AuNPs' plasmon characteristics within 30 min of antibody introduction. Co-immobilization of two epitopes improved the assay sensitivity relative to single-epitope AuNPs with a limit of detection of 3.2 nM, commensurate with IgG levels in convalescent COVID-19-infected patients. A passivation strategy was further pursued to preserve the sensing response in human plasma medium. When tested against 35 clinical plasma samples of varying illness severity, the optimized nanosensor assay can successfully identify SARS-CoV-2 infection with 100% specificity and 83% sensitivity. As the epitopes are conserved within the circulating COVID-19 variants, the proposed platform holds great potential to serve as a cost-effective and highly specific alternative to classical immunoassays employing recombinant viral proteins. These epitope-enabled nanosensors further expand the serodiagnostic toolbox for COVID-19 epidemiological study, humoral response monitoring, or vaccine efficiency assessment.

Lithographic Processes for the Scalable Fabrication of Micro- and Nanostructures for Biochips and Biosensors.

Since the early 2000s, extensive research has been performed to address numerous challenges in biochip and biosensor fabrication in order to use them for various biomedical applications. These biochips and biosensor devices either integrate biological elements (e.g., DNA, proteins or cells) in the fabrication processes or experience post fabrication of biofunctionalization for different downstream applications, including sensing, diagnostics, drug screening, and therapy. Scalable lithographic techniques that are well established in the semiconductor industry are now being harnessed for large-scale production of such devices, with additional development to meet the demand of precise deposition of various biological elements on device substrates with retained biological activities and precisely specified topography. In this review, the lithographic methods that are capable of large-scale and mass fabrication of biochips and biosensors will be discussed. In particular, those allowing patterning of large areas from 10 cm2 to m2, maintaining cost effectiveness, high throughput (>100 cm2 h-1), high resolution (from micrometer down to nanometer scale), accuracy, and reproducibility. This review will compare various fabrication technologies and comment on their resolution limit and throughput, and how they can be related to the device performance, including sensitivity, detection limit, reproducibility, and robustness.