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Development of a ZnCdS@ZnS quantum dotsa??based label-free electrochemiluminescence immunosensor for sensitive determination of aflatoxin B1 in lotus seed
摘要： In this study, we designed a ZnCdS@ZnS quantum dots (QDs)–based label-free electrochemiluminescence (ECL) immunosensor for sensitive determination of aflatoxin B1 (AFB1). A Nafion solution assembled abundant QDs on the surface of a Au electrode as ECL signal probes, with specially coupled anti-AFB1 antibodies as the capturing element. As the reduction reaction between S2O8 2? in the electrolyte and QDs on the electrode led to ECL emission, the decreased ECL signals resulting from target AFB1 in the samples were recorded for quantification. We evaluated electrochemical impedance spectroscopy and ECL measurements along each step in the construction of the proposed immunosensor. After systematic optimization of crucial parameters, the ECL immunosensor exhibited a good sensitivity, with a low detection limit of 0.01 ng/mL for AFB1 in a wide concentration range of 0.05–100 ng/mL. Testing with lotus seed samples confirmed the satisfactory selectivity, stability, and reproducibility of the developed ECL immunosensor for rapid, efficient, and sensitive detection of AFB1 at trace levels in complex matrices. This study provides a powerful and universal analytical platform for a variety of analytes that can be used in broad applications for real-time analysis, such as food and traditional Chinese medicine safety testing, environmental pollution monitoring, and disease diagnostics.
关键词： Electrochemiluminescence immunosensor,Nafion,Lotus seed,Aflatoxin B1,ZnCdS@ZnS quantum dots,Label-free
更新于2025-09-23 15:19:57
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Mechanisms of Fano-resonant biosensing: Mechanical loading of plasmonic oscillators
摘要： Distinctively narrow and asymmetric line shape Fano resonances arise due to resonant interactions of sub-radiant and super-radiant modes in plasmonic nanostructures and metamaterials. A number of recent experimental studies have shown unique opportunities provided by highly dispersive Fano resonances in biosensing applications. However, there is limited understanding of Fano resonant optical response to biomolecular accumulation. Here, we introduce a phenomenological model that can precisely describe the intricate nature of the Fano resonances in plasmonic nanohole arrays and provide unambiguous physical insights into biosensing experiments. Using rigorous electromagnetic simulations and experimental measurements as benchmarking tools, we show that the non-trivial contribution of molecular accumulation to Fano resonant plasmonic response can be incorporated as a mechanical loading effect in a coupled-oscillator model. Quite remarkably, our phenomenological approach captures the complex spectral response of the Fano resonance profile and asymmetric linewidth broadening upon molecular accumulation. Furthermore, in strong agreement with our experimental measurements, we show that our parameterized model has predictive power in fine tuning the Fano resonant extraordinary light transmission lineshape using structural design parameters without resorting to electromagnetic simulations. Our phenomenological model provides a general analytical method that can be adapted to understand biomolecular detection measurements in different plasmonic and metamaterial systems.
关键词： Biosensing,Extraordinary light transmission,Plasmonics,Plasmonic nanoholes,Fano resonances,Label-free detection
更新于2025-09-23 15:19:57
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A Nanostructured Gold/Graphene Microfluidic Device for Direct and Plasmonic-Assisted Impedimetric Detection of Bacteria
摘要： Hierarchical 3D gold nano-micro islands (NMIs) are favourably structured for direct and probe-free capture of bacteria in optical and electrochemical sensors. Moreover, their unique plasmonic properties make them a suitable candidate for plasmonic-assisted electrochemical sensors, yet the charge transfer needs to be improved. In the present study, we propose a novel plasmonic-assisted electrochemical impedimetric detection platform based on hybrid structures of 3D gold NMIs and graphene (Gr) nanosheets for probe-free capture and label-free detection of bacteria. The inclusion of Gr nanosheets significantly improves the charge transfer, addressing the central issue of using 3D gold NMIs. Notably, the 3D gold NMIs/Gr detection platform successfully distinguishes between various types of bacteria including Escherichia coli (E. coli) K12, Pseudomonas putida (P. putida) and Staphylococcus epidermidis (S. epidermidis) when electrochemical impedance spectroscopy is applied under visible light. We show that distinguishable and label-free impedimetric detection is due to dissimilar electron charge transfer caused by various sizes, morphologies, and compositions of the cells. In addition, the finite-difference time-domain (FDTD) simulation of the electric field indicates the intensity of charge distribution at the edge of the NMI structures. Furthermore, the wettability studies demonstrated that contact angle is a characteristic feature of each type of captured bacteria on the 3D gold NMIs, which strongly depends on the shape, morphology, and size of the cells. Ultimately, exposing the platform to various dilutions of the three bacteria strains, revealed the ability to detect dilutions as low as ~20 CFU/mL in a wide linear range of detection of 2(cid:3)101-105, 2(cid:3)101-104 and 1(cid:3)102-1(cid:3)105 CFU/mL for E. coli, P. putida, and S. epidermidis, respectively. The proposed hybrid structure of 3D gold NMIs and Gr combined by novel plasmonic and conventional impedance spectroscopy techniques open interesting avenues in ultrasensitive label-free detection of bacteria with low cost and high stability.
关键词： Label-free bacteria detection,Hierarchical gold nano-micro islands,Surface properties,Impedance spectroscopy,plasmonic-assisted electrochemical detection platform
更新于2025-09-23 15:19:57
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Fungal In Situ Assembly Gives Novel Properties to CdS <sub/><i>x</i> </sub> Se <sub/> 1a?? <i>x</i> </sub> Quantum Dots for Sensitive Label-Free Detection of Chloramphenicol
摘要： Quantum dots (QDs) are attracting significant attention for the development of sensitive detection methods because of their unique optical properties. Biosynthetic QDs using organisms is a promising alternative route to chemical synthesis because it is a simple and eco-friendly process. Herein, we developed a straightforward and green system for the biological assembly of CdSxSe1?x QDs by Phomopsis sp. XP-8 within 6 h, a more efficient approach than those reported in other organisms. The QDs were shown to have a CdS0.75Se0.25@oligopeptide transporter structure, and the production process was shown to be strongly influenced by intracellular glutathione content. The QDs were monodispersed with a uniform spherical shape of 3.22 ± 0.07 nm in diameter. They exhibited good water solubility and excellent fluorescence properties. The QDs could be extracted and used directly as a sensitive chloramphenicol (CAP) probe via static fluorescence quenching in the linear range from 3.13 to 500 μg/L with a detection limit of 0.89 μg/L. The detection method was highly selective for CAP with minimal interference from other antibiotics and was used to successfully detect CAP in milk samples. Overall, this work has great significance for the development of a fast and simple QD synthesis system via biological assembly.
关键词： Phomopsis sp. XP-8,Chloramphenicol,CdSxSe1?x quantum dots,Biosynthesis,Label-free detection
更新于2025-09-23 15:19:57
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DNA-MnO2 Nanosheets as Washing- and Label-Free Platform for Array-Based Differentiation of Cell Types
摘要： Accurate and facile differentiation of cell types is critical for accurate diagnosis and therapy of diseases. However, it remains challenging due to low specificity, requirement of sophisticated instruments, and tedious operation steps. Herein, a simple, washing- and label-free chemical tongue was constructed for differentiation of cell types. In the array-based sensing platform, DNA-ligand ensembles adsorbed on the surface of MnO2 nanosheets were used as sensing probes. Instead of aptamers from cell-SELEX, the randomly designed DNA strands were used, offering versatile interactions with cells. The property that MnO2 nanosheets can be degraded by intracellular glutathione makes the platform avoid the washing step. Eight types of cell lines were distinguished from each other after the data were treated with principal component analysis (PCA). In addition, a 95% of identification accuracy for the randomly selected unknown samples was achieved. The strategy shows an excellent performance not only in distinguishing cell lines but also in the identification of unknown cell samples.
关键词： label-free,pattern recognition,cell types,DNA-MnO2 nanosheets,washing-free
更新于2025-09-19 17:15:36
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Principles for Sensitive and Robust Biomolecular Interaction Analysis: The Limits of Detection and Resolution of Diffraction-Limited Focal Molography
摘要： Label-free biosensors enable the monitoring of biomolecular interactions in real time, which is key to the analysis of the binding characteristics of biomolecules. While refractometric optical biosensors such as surface plasmon resonance (SPR) are sensitive and well-established, they are susceptible to any change of the refractive index in the sensing volume caused by minute variations in composition of the sample buffer, temperature drifts, and most importantly nonspecific binding to the sensor surface in complex fluids such as blood. The limitations arise because refractometric sensors measure the refractive index of the entire sensing volume. Conversely, diffractometric biosensors–for example, focal molography–only detect the diffracted light from a coherent assembly of analyte molecules. Thus any refractive index distribution that is noncoherent with respect to this molecular assembly does not add to the coherent signal. This makes diffractometric biosensors inherently robust and enables sensitive measurements without reference channels or temperature stabilization. The coherent assembly is generated by selective binding of the analyte molecules to a synthetic binding pattern–the mologram. Focal molography has been introduced theoretically [C. Fattinger, Phys. Rev. X 4, 031024 (2014)] and verified experimentally [V. Gatterdam, A. Frutiger, K.-P. Stengele, D. Heindl, T. Lübbes, J. V?r?s, and C. Fattinger, Nat. Nanotechnol. 12, 1089 (2017)] in previous papers. However, further understanding of the underlying physics and a diffraction-limited readout is needed to unveil its full potential. This paper introduces refined theoretical models, which can accurately quantify the amount of biological matter bound to the mologram from the diffracted intensity. In addition, it presents measurements of diffraction-limited molographic foci, i.e., Airy discs. These improvements enable us to demonstrate a resolution in real-time binding experiments comparable to the best SPR sensors without the need for temperature stabilization or drift correction and to detect low-molecular-weight compounds label free in an endpoint format. The presented experiments exemplify the robustness and sensitivity of the diffractometric sensor principle.
关键词： biomolecular interaction analysis,diffractometric biosensors,focal molography,diffraction-limited foci,robustness,label-free detection,sensitivity
更新于2025-09-19 17:15:36
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High-throughput label-free molecular fingerprinting flow cytometry
摘要： Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm?1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics of Haematococcus lacustris.
关键词： high-throughput,astaxanthin,label-free,single-cell analysis,microfluidics,flow cytometry,Raman spectroscopy,Haematococcus lacustris,coherent anti-Stokes Raman scattering
更新于2025-09-19 17:15:36
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Protein-functionalized WO3 nanorods–based impedimetric platform for sensitive and label-free detection of a cardiac biomarker
摘要： We report the development of a sensitive and a label-free electrochemical immunosensing platform for the detection of cardiac biomarker troponin I (cTnI) using tungsten trioxide nanorods (WO3 NRs). The low-temperature hydrothermal technique was employed for the controlled synthesis of WO3 NRs. Thin films of 3-aminopropyltriethoxy saline (APTES)-functionalized WO3 NRs were deposited on indium tin oxide (ITO)-coated glass substrate (0.5 cm × 1 cm) using electrophoretic deposition technique. The covalent immobilization of cTnI antibody onto functionalized WO3 NRs electrode was accomplished using EDC-NHS [1-(3-(dimethylamino)-propyl)-3-ethylcarbodiimide hydrochloride and N-hydroxysulfosuccinimide] chemistry. The structural and morphological characterizations of WO3 NRs and functionalized WO3 NRs were studied using X-ray diffraction, field emission scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and electrochemical techniques. The impedimetric response study of the proposed immunosensor demonstrates high sensitivity [6.81 KΩ·mL/(ng·cm2)] in a linear detection range of 0.01–10 ng/mL. The excellent selectivity, good reproducibility, and long-term stability of the proposed immunosensing platform indicate WO3 NRs as a suitable platform for the development of a point-of-care biosensing device for cardiac detection.
关键词： electrochemical impedance spectroscopy,tungsten trioxide nanorods,immunosensor,cardiac biomarker,label-free detection
更新于2025-09-19 17:15:36
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Label-free fluorescence assay coupled exonuclease reaction and SYBR Green I for the detection of T4 polynucleotide kinase activity
摘要： A sensitive label-free ?uorescence assay for monitoring T4 polynucleotide kinase (T4 PNK) activity and inhibition was developed based on a coupled l exonuclease cleavage reaction and SYBR Green I. In this assay, a double-stranded DNA (dsDNA) was stained with SYBR Green I and used as a substrate for T4 PNK. After the 50-hydroxyl termini of the dsDNA was phosphorylated by the T4 PNK, the coupled l exonuclease began to digest the dsDNA to form mononucletides and single-stranded DNA (ssDNA). At this moment, the ?uorescence intensity of the SYBR Green I decreased because of less a?nity with ssDNA than dsDNA. The decreasing extent was proportional to the concentration of the T4 PNK. After optimization of the detection conditions, including the concentration of ATP, amount of l exonuclease and reaction time, the activity of T4 PNK was monitored by the ?uorescence measurement, with the limit of detection of 0.11 U mL(cid:2)1 and good linear correlation between 0.25–1.00 U mL(cid:2)1 (R2 ? 0.9896). In this assay, no label was needed for ?uorescence detection. Moreover, the inhibition behaviors of the T4 PNK's inhibitors were evaluated by this assay. The result indicated the potential of using this assay for monitoring of the phosphorylation-related process.
关键词： SYBR Green I,label-free ?uorescence assay,T4 polynucleotide kinase activity,exonuclease reaction
更新于2025-09-19 17:13:59
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OPTICAL BIOSENSORS PROSPECTIVE BASED ON BRAGG GRATING POLYMER WAVEGUIDE
摘要： In this work, we demonstrate the potential of Bragg grating polymer waveguide as an optical biosensor. Visible wavelength region at 650 nm is used as a centre wavelength because it is commonly used in biological and chemical sensing for both label and label-free sensing. The Bragg polymer waveguide structure is simulated using RSoft optical design and analysis software. The results show that there is a transmission drop with a 3 dB bandwidth of 661.0 nm when the surrounding refractive index is 1.33. The specific wavelength (transmission drop) is shifted to 724.2 nm when we increased the surrounding medium into 1.43 to mimic the bioanalytes solution. Simulation result shows that the wavelength shift was approximately 63.2 nm for every 0.1 increasing of surrounding refractive index. The Bragg grating polymer waveguide was fabricated by using electron beam lithography. Then, the fabricated devices were easily integrated within microfluidic systems in order to validate the wavelength shift. From the experiments, the wavelength shift occurred approximately 20.3 nm over 0.1 increment of refractive index. The discrepancies were likely due to the accumulation of sucrose solution on top and sidewall of the sensing area, the insertion loss between input and output coupling of the waveguide interface that induced the noise to signal ratio. Where we know that, is impossible to happen in simulation. Thus both simulation and experimental results strongly indicate that Bragg grating polymer waveguide structure at visible wavelength region have a potential for label or label-free optical biosensing applications.
关键词： simulation,label-free sensing,polymer waveguide,Optical biosensor,Bragg grating
更新于2025-09-19 17:13:59