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- 摘要
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- 实验方案
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Invisible-ink-assisted pattern and written surface-enhanced Raman scattering substrates for versatile chem/biosensing platforms
摘要: In recent years, highly sensitive surface-enhanced Raman scattering (SERS) integrated with flexible substrates has drawn increasing attention for label-free detection. In this study, an invisible ink-inspired process was developed for the fabrication of plasmonic Au-based SERS substrates through an on-site redox strategy. Tannic acid (TNA), a common green reagent, was used not only for fabricating various SERS absorbents through a confinement reduction of a Au-TNA complex, but also for supplying an amphiphilic inorganic–organic surface structure for outstanding SERS enhancement at micromolar to nanomolar concentrations for a wide range of compounds. In addition to label-free sensing, this TNA/Au-based SERS substrate provides a versatile analysis platform for studies of chemical and biological reactions. A combination of TNA ink with different metal ions allows for a reliable procedure for the synthesis of a bimetallic AuAg SERS substrate that further enhances the SERS intensity of analyte molecules and extends the lower limit of detection.
关键词: bimetallic AuAg SERS substrate,invisible ink,tannic acid,label-free detection,SERS,plasmonic Au-based SERS substrates,surface-enhanced Raman scattering
更新于2025-09-23 15:21:21
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Live E. coli bacteria label-free sensing using a microcavity in-line Mach-Zehnder interferometer
摘要: The paper presents the first study to date on selective label-free biosensing with a microcavity in-line Mach-Zehnder interferometer induced in an optical fiber. The sensing structures were fabricated in a single-mode fiber by femtosecond laser micromachining. In contrast to other studies of this sensing scheme, where only the sensitivity to refractive index changes in the cavity was investigated, this research used chemical surface treatment of the sensor to ensure detection specificity. Immobilized MS2 bacteriophages were applied as recognition elements specifically targeting live E. coli C3000 bacteria. It is shown that the sensor allows for real-time monitoring of biological phenomena taking place on the surface of the microcavity. The developed biosensor exhibits ultrahigh refractive index sensitivity of 15,000 nm/RIU and is capable of detecting live E. coli bacteria concentrations as low as 100 colony forming units (CFU)/mL in liquid volume as low as picoliters.
关键词: label-free biosensing,E. coli C3000 bacteria,refractive index sensitivity,MS2 bacteriophages,femtosecond laser micromachining,microcavity in-line Mach-Zehnder interferometer,optical fiber
更新于2025-09-23 15:21:21
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Label-Free Imaging of Single Proteins Secreted from Living Cells via iSCAT Microscopy
摘要: We demonstrate interferometric scattering (iSCAT) microscopy, a method capable of detecting single unlabeled proteins secreted from individual living cells in real time. In this protocol, we cover the fundamental steps to realize an iSCAT microscope and complement it with additional imaging channels to monitor the viability of a cell under study. Following this, we use the method for real-time detection of single proteins as they are secreted from a living cell which we demonstrate with an immortalized B-cell line (Laz388). Necessary steps concerning the preparation of microscope and sample as well as the analysis of the recorded data are discussed. The video protocol demonstrates that iSCAT microscopy offers a straightforward method to study secretion at the single-molecule level.
关键词: imaging,single-protein,real-time,scattering,dynamics,cellular secretion,label-free,iSCAT
更新于2025-09-23 15:21:21
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Optoplasmonic characterisation of reversible disulfide interactions at single thiol sites in the attomolar regime
摘要: Probing individual chemical reactions is key to mapping reaction pathways. Trace analysis of sub-kDa reactants and products is obfuscated by labels, however, as reaction kinetics are inevitably perturbed. The thiol-disulfide exchange reaction is of specific interest as it has many applications in nanotechnology and in nature. Redox cycling of single thiols and disulfides has been unresolvable due to a number of technological limitations, such as an inability to discriminate the leaving group. Here, we demonstrate detection of single-molecule thiol-disulfide exchange using a label-free optoplasmonic sensor. We quantify repeated reactions between sub-kDa thiolated species in real time and at concentrations down to 100’s of attomolar. A unique sensing modality is featured in our measurements, enabling the observation of single disulfide reaction kinetics and pathways on a plasmonic nanoparticle surface. Our technique paves the way towards characterising molecules in terms of their charge, oxidation state, and chirality via optoplasmonics.
关键词: Optoplasmonic,Label-free sensing,Attomolar regime,Thiol-disulfide exchange,Single-molecule detection
更新于2025-09-23 15:21:01
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Infrared spectroscopy of live cells from a flowing solution using electrically-biased plasmonic metasurfaces
摘要: Spectral cytopathology (SCP) is a promising label-free technique for diagnosing diseases and monitoring therapeutic outcomes using FTIR spectroscopy. In most cases, cells must be immobilized on a substrate prior to spectroscopic interrogation. This creates significant limitations for high throughput phenotypic whole-cell analysis, especially for the non-adherent cells. Here we demonstrate how metasurface-enhanced infrared reflection spectroscopy (MEIRS) can be applied to a continuous flow of live cell solution by applying AC voltage to metallic metasurfaces. By integrating metasurfaces with microfluidic delivery channels and attracting the cells to the metasurface via dielectrophoretic (DEP) force, we collect the infrared spectra of cells in real time within a minute, and correlate the spectra with simultaneously acquired images of the attracted cells. The resulting DEP-MEIRS technique paves the way for rapid SCP of complex cell-containing body fluids with low cell concentrations, and for the development of a wide range of label-free liquid biopsies.
关键词: dielectrophoretic force,live cells,FTIR spectroscopy,microfluidic delivery channels,Spectral cytopathology,metasurface-enhanced infrared reflection spectroscopy,label-free liquid biopsies
更新于2025-09-23 15:21:01
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Setting Up Surface-Enhanced Raman Scattering Database for Artificial Intelligence-Based Label-Free Discrimination of Tumor Suppressor Genes
摘要: The quality of input data in deep learning is tightly associated with the ultimate performance of machine learner. Taking advantages of unique merits of surface-enhanced Raman scattering (SERS) methodology in the collection and construction of database (e.g., abundant intrinsic fingerprint information, noninvasive data acquisition process, strong anti-interfering ability, etc.), herein we set up SERS-based database of deoxyribonucleic acid (DNA), suitable for artificial intelligence (AI)-based sensing applications. The database is collected and analyzed by silver nanoparticles (Ag NPs)-decorated silicon wafer (Ag NPs@Si) SERS chip, followed by training with a deep neural network (DNN). As proof-of-concept applications, three kinds of representative tumor suppressor genes, i.e., p16, p21 and p53 fragments, are readily discriminated in label-free manners. Prominent and reproducible SERS spectra of these DNA molecules are collected and employed as input data for DNN learning and training, which enables selective discrimination of DNA target(s). The accuracy rate for the recognition of specific DNA target reaches 90.28%.
关键词: surface-enhanced Raman scattering,label-free discrimination,deep neural network,tumor suppressor genes,artificial intelligence
更新于2025-09-23 15:21:01
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Label-Free Biosensing Method for the Detection of a Pancreatic Cancer Biomarker Based on Dielectrophoresis Spectroscopy
摘要: We show that negative dielectrophoresis (DEP) spectroscopy is an effective transduction mechanism of a biosensor for the diagnosis and prognosis of pancreatic cancer using the biomarker CA 19-9. A substantial change in the negative DEP force applied to functionalized polystyrene microspheres (PM) was observed with respect to both the concentration level of the pancreatic cancer biomarker CA 19-9 and the frequency of the electric field produced by a pearl shaped interdigitated gold micro-electrode. The velocity of repulsion of a set of PM functionalized to a monoclonal antibody to CA 19-9 was calculated for several concentration cutoff levels of CA 19-9, including 0 U/mL and 37 U/mL, at the frequency range from 0.5 to 2 MHz. The velocity of repulsion of the PM from the electrode was determined using a side illumination and an automated software using a real-time image processing technique that captures the Mie scattering from the PM. Since negative DEP spectroscopy is an effective transduction mechanism for the detection of the cutoff levels of CA 19-9, it has the potential to be used in the early stage diagnosis and in the prognosis of pancreatic cancer.
关键词: spectroscopy,pancreatic cancer,dielectrophoresis,label free,bioelectronics
更新于2025-09-23 15:21:01
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EDC-Mediated Oligonucleotide Immobilization on a Long Period Grating Optical Biosensor
摘要: We present the development and simplification of label-free fiber optic biosensors based on immobilization of oligonucleotides on dual-peak long period gratings (dLPGs). This improvement is the result of a simplification of biofunctionalization methodology. A one-step 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)-mediated reaction has been developed for the straightforward immobilization of unmodified oligonucleotides on the glass fiber surface along the grating region, leading to covalent attachment of a 5′-phosphorylated probe oligonucleotide to the amino-derivatized fiber grating surface. Immobilization is achieved via a 5′phosphate-specific linkage, leaving the remainder of the oligonucleotide accessible for binding reactions. The dLPG has been tested in different external media to demonstrate its inherent ultrahigh sensitivity to the surrounding-medium refractive index (RI) achieving 50-fold improvement in RI sensitivity over the previously-published LPG sensor in media with RI’s relevant to biological assays. After functionalization, the dLPG biosensor was used to monitor the hybridization of complementary oligonucleotides showing a detectable oligonucleotide concentration of 4 nM. The proposed one-step EDC reaction approach can be further extended to develop fiber optic biosensors for disease analysis and medical diagnosis with the advances of label-free, real-time, multiplex, high sensitivity and specificity.
关键词: EDC-mediated reaction,Label-free,DNA,Oligonucleotide,Optical fiber biosensor,Long period grating
更新于2025-09-23 15:21:01
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Highly selective and label-free Love-mode surface acoustic wave biosensor for carcinoembryonic antigen detection using a self-assembled monolayer bioreceptor
摘要: A love-mode surface acoustic wave (SAW) biosensor based on ST-cut quartz was developed for highly selective and label-free detection of carcinoembryonic antigen (CEA). The delay line area of an interdigital transducer (IDT) based SAW device was coated with gold and then chemically modified through thioglycolic acid–EDC/NHS reaction mechanism. A self-assembled monolayer of anti-CEA was further immobilized on the bioreceptors through the coupling layer. The biosensing capability of the SAW device was evaluated using solutions of CEA with various concentrations and limit of detection was obtained at 0.31 ng/ml of CEA, which is better than the results reported by the literatures available for CEA detection using SAW device. The real-time detection capability of the biosensor was evaluated using clinical serum samples and selectivity was evaluated using mixed solutions of CEA with other common tumor marking proteins. Long-term stability of the biosensor was also evaluated over a period of 30 days and the immunoassay response has shown only 8% decrease in performance within the whole period. The binding of CEA onto the bioreceptor was evaluated through Langmuir and Freundlich sorption isotherm kinetic studies as well.
关键词: SAW,Piezoelectricity,Biosensing,CEA,Label free,SAM
更新于2025-09-23 15:19:57
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A fully electronically tunable millimeter wave lab-in-waveguide nano-fluidic sensor
摘要: In this paper, we report an ultra-sensitive millimeter wave (mm-wave) reflection liquid sensor, which is electronically tunable, non-invasive, and label-free. The reflection liquid sensor is based on the impedance match technique. It is implemented in a rectangular waveguide structure, which makes the interaction between mm-wave and liquid under test (LUT) in a closed environment, forms a nano-fluidic lab-in-waveguide liquid sensor, and isolates the interference of surroundings. The reflection liquid sensor shows high sensitivity to permittivity change of the LUTs. Taking the ethanol-distilled water as benchmark, a more than 62 dB ethanol-distilled water contrast is experimentally achieved. The detection limit is 0.05% ethanol in volumetric concentration with a 4 dB signal difference, which is the best performance of the microwave and mm-wave liquid sensors publicly reported until now. The ultra-sensitive reflection sensor can be used for bio-chemical sensing. The lab-in-waveguide biosensor is also compatible with microfluidics.
关键词: impedance match,electronically tunable,liquid sensor,label-free,millimeter wave,non-invasive
更新于2025-09-23 15:19:57