The ongoing global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has led to active research in its associated diagnostics and medical treatments. While quantitative reverse transcription polymerase chain reaction (qRT-PCR) is the most reliable method to detect viral genes of SARS-CoV-2, serological tests for specific antiviral antibodies are also important as they identify false negative qRT-PCR responses, track how effectively the patient’s immune system is fighting the infection, and are potentially helpful for plasma transfusion therapies. In this work, based on the principle of localized surface plasmon resonance (LSPR), we develop an opto-microfluidic sensing platform with gold nanospikes, fabricated by electrodeposition, to detect the presence and amount of antibodies specific to the SARS-CoV-2 spike protein in 1 L of human plasma diluted in 1mL of buffer solution, within 30min. The target antibody concentration can be correlated with the LSPR wavelength peak shift of gold nanospikes caused by the local refractive index change due to the antigen–antibody binding. This label-free microfluidic platform achieves a limit of detection of 0.08ng/mL ( 0.5pM), falling under the clinical relevant concentration range. We demonstrate that our opto-microfluidic platform offers a promising point-of-care testing tool to complement standard serological assays and make SARS-CoV-2 quantitative diagnostics easier, cheaper, and faster.
DNA polymerase catalyzes the replication of DNA, one of the key steps in cell division. The control and understanding of this reaction owns great potential for the fundamental study of DNA-enzyme interactions. In this context, we developed a label-free microfluidic biosensor platform based on the principle of localized surface plasmon resonance (LSPR) to detect the DNA-polymerase reaction in real-time. Our microfluidic LSPR chip integrates a polydimethylsiloxane (PDMS) channel bonded with a nanoplasmonic substrate, which consists of densely packed mushroom-like nanostructures with silicon dioxide stems (~40 nm) and gold caps (~22 nm), with an average spacing of 19 nm. The LSPR chip was functionalized with single-stranded DNA (ssDNA) template (T30), spaced with hexanedithiol (HDT) in a molar ratio of 1:1. The DNA primer (P8) was then attached to T30, and the second strand was subsequently elongated by DNA polymerase assembling nucleotides from the surrounding fluid. All reaction steps were detected in-situ inside the microfluidic LSPR chip, at room temperature, in real-time, and label-free. In addition, the sensor response was successfully correlated with the amount of DNA and HDT molecules immobilized on the LSPR sensor surface. Our platform represents a benchmark in developing microfluidic LSPR chips for DNA-enzyme interactions, further driving innovations in biosensing technologies.
The recent advances in nanofabrication processes offer encouraging opportunities for designing highly sensitive detection tools. One example is metal-enhanced fluorescence (MEF)-based biosensors: by effectively coupling the metal nanostructures with the fluorescent dye used for the detection of the target molecule, MEF-based sensors exhibit higher sensitivity and lower limit of detection in comparison to traditional optical biosensors. Ordered arrays of nanostructures with coupled fluorophores can potentially achieve thousand-fold enhancement in fluorescence intensity. However, nanofabrication techniques required for ordered nanostructures tend to be time consuming and costly. On the other hand, with moderate enhancement, randomly assembled nanoplasmonic arrays on large-scale substrates can be realized by easy, cost-effective, and reliable methodologies in a relatively short time, thus being more suitable for mass production. In this work, we develop a MEF-based immunosensor involving randomly assembled plasmonic arrays of gold nanoparticles fabricated by a simple and versatile three-step process to modulate the size, the interparticle distance, and the optical properties of the gold nanostructures to achieve the optimal fluorescence enhancement. Specifically, we have tested three different fluorescent dyes (Alexa Fluor 488, Alexa Fluor 546, and PE-Cy7) coupled with optimized gold nanostructures and achieved up to ≈170-fold enhancement in the fluorescence emission, significantly better than those achieved by metal nanostructures in solutions and by randomly assembled nanoplasmonic arrays. Our MEF immunosensor is then employed for detecting immunoglobulins in a model antigen−antibody system, achieving a limit of detection of 4.3 ng/mL. This value is lower than that of similar immunoglobulin detection assays and offers promising opportunities for a wide range of biosensing applications.
Microbial biofilms possess intrinsic resistance against conventional antibiotics and cleaning procedures; thus, a better understanding of their complex biological structures is crucial in both medical and industrial applications. Existing laboratory methodologies have focused on macroscopic and mostly indirect characterization of mechanical and microbiological properties of biofilms adhered on a given substrate. However, the kinetics underlying the biofilm formation is not well understood, while such information is critical to understanding how drugs and chemicals influence the biofilm formation. Herein, we report the use of localized surface plasmon resonance (LSPR) for real-time, label-free monitoring of E. coli biofilm assembly on a nanoplasmonic substrate consisting of gold mushroom-like structures. Our LSPR sensor is able to capture the signatures of biofilm formation in real-time by measuring the wavelength shift in the LSPR resonance peak with high temporal resolution. We employ this sensor feature to elucidate how biofilm formation is affected by different drugs, including conventional antibiotics (kanamycin and ampicillin) as well as rifapentine, a molecule preventing cell adhesion yet barely affecting bacterial viability and vitality. Due to its flexibility and simplicity, our LSPR based platform can be used on a wide variety of clinically relevant bacteria, thus representing a valuable tool in biofilm characterization and drug screening.
In this paper, the interesting hydrogen sensing properties of a Pd-gate AlGaN/GaN Schottky diode are investigated. A significantly low detection limit of 850 ppb H2/air gas can be observed with increasing the temperature to 423 K. The experimental results indicate that hydrogen molecules cause great influences on the diode breakdown voltage. Also, the diode exhibits an ultrahigh sensing response of 2.04×105 at 423 K when exposure to a 9660 ppm H2/air gas. The transient response time and reversibility of the studied device can be improved by increasing the operating temperature.
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