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Thin-core fiber-optic biosensor for DNA hybridization detection
摘要： A real-time label-free DNA biosensor based on thin-core fiber (TCF) interferometer is demonstrated experimentally. The proposed biosensor is constructed by splicing a TCF between two segments of single mode fibers (SMFs) and integrated into a microfluidic channel. By modifying the TCF surface with monolayer poly-l-lysine (PLL) and single-stranded deoxyribonucleic acid (ssDNA) probes, the target DNA molecules can be captured in the microfluidic channel. The transmission spectra of the biosensor are measured and theoretically analyzed under different biosensing reaction processes. The results show that the wavelength has a blue-shift with the process of the DNA hybridization. Due to the advantages of low cost, simple operation as well as good detection effect on DNA molecules hybridization, the proposed biosensor has great application prospects in the fields of gene sequencing, medical diagnosis, cancer detection and environmental engineering.
关键词： thin-core fiber,biosensor,microfluidic channel,modal interference,DNA hybridization
更新于2025-11-28 14:23:57
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Mapping Neurotransmitter Identity in the Whole-Mount <i>Drosophila</i> Brain Using Multiplex High-Throughput Fluorescence <i>in Situ</i> Hybridization
摘要： Identifying the neurotransmitters used by specific neurons is a critical step in understanding the function of neural circuits. However, methods for the consistent and efficient detection of neurotransmitter markers remain limited. Fluorescence in situ hybridization (FISH) enables direct labeling of type-specific mRNA in neurons. Recent advances in FISH allow this technique to be carried out in intact tissue samples such as whole-mount Drosophila melanogaster brains. Here, we present a FISH platform for high-throughput detection of eight common neurotransmitter phenotypes in Drosophila brains. We greatly increase FISH throughput by processing samples mounted on coverslips and optimizing fluorophore choice for each probe to facilitate multiplexing. As application examples, we demonstrate cases of neurotransmitter co-expression, reveal neurotransmitter phenotypes of specific cell types and explore the onset of neurotransmitter expression in the developing optic lobe. Beyond neurotransmitter markers, our protocols can in principle be used for large scale FISH detection of any mRNA in whole-mount fly brains.
关键词： Neurotransmitter,mRNA,Fluorescence in situ hybridization,Gene expression,Drosophila
更新于2025-11-21 11:08:12
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Ultrathin Ti3C2 Nanosheets based “off-on” Fluorescent nanoprobe for Rapid and Sensitive Detection of HPV Infection
摘要： MXenes as a new class of 2D materials have recently been widely applied in energy storage, electrocatalysis, sensors, adsorption, water purification, and so on, due to their tunable versatile properties. Herein, we demonstrate a simple, rapid and highly-sensitive sensing platform based on ultrathin two-dimensional MXene Ti3C2 nanosheets (Ti3C2 NSs) for selective analysis of Human papillomavirus (HPV), a major human pathogens and causative agents of cervical cancer. Ultrathin Ti3C2 NSs, obtained by exfoliating their layered HF-etched powder, exhibit high fluorescence quenching ability to dye-labeled single-stranded DNA (ssDNA) and different affinities for ssDNA and double-stranded DNA (dsDNA). Under the fluorescence quenching effect of Ti3C2 NSs, ssDNA probe (P) shows the minimal fluorescent emission. After the formation of duplex structure with its complementary target, ssDNA (T), the fluorescence intensity enhances evidently. Exonuclease III (Exo III) was used to improve the sensitivity by promoting more fluorescence enhancement. This magnified fluorescent sensor for HPV-18 detection shows a low detection limit of 100 pM and a high specificity. Furthermore, the developed DNA sensor can be employed to determine PCR amplified HPV-18 from cervical scrapes samples. It highlights ultrathin Ti3C2 NSs as a potential candidate for construction of fluorescence DNA biosensors with excellent performances.
关键词： DNA hybridization,Ti3C2 nanosheets,HPV,Fluorescent detection
更新于2025-11-19 16:46:39
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Enzyme-free “on-off-on” photoelectrochemical biosensor based on cascaded quadratic amplification strategy for miRNA 141 detection
摘要： MicroRNAs (miRNAs) assay is of great significance for early diagnosis of diseases, so an enzyme-free “on-off-on” PEC biosensor has been developed for sensitive miRNA 141 determination. Manganese-doped cadmium sulfide coupled with zinc sulfide quantum dots (Mn:CdS@ZnS QDs) and manganese porphyrin (MnPP) have been used as photoelectric material and photosensitizer, respectively. And a high photocurrent of approximately 70.0 μA has been obtained. Cascaded quadratic amplification strategy has been applied in the system. Mn:CdS@ZnS QDs was characterized by transmission electron microscopy (TEM), scanning electron microscope (SEM), X-ray photoelectron spectroscopy (XPS) and energy dispersive X-ray spectroscopy (EDX). Photoelectrochemical and electrochemical technologies were used to monitor the fabrication process of the biosensor. The sensing platform exhibits recommendable stability and good selectivity, miRNA 141 can be accurately quantified with a linear range of 1.00 × 10-14 to 1.00 × 10-8 mol·L-1 and the detection limit of 3.30 fmol·L-1. This method provides promising potential to explore sensitive detection models for various biological molecules.
关键词： Hybridization chain reaction,Catalytic hairpin assembly,Manganese-doped cadmium sulfide coupled with zinc sulfide quantum dots,MiRNA 141,Photoelectrochemistry,On-off-on
更新于2025-11-14 17:03:37
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Reproduction of surface-enhanced resonant Raman scattering and fluorescence spectra of a strong coupling system composed of a single silver nanoparticle dimer and a few dye molecules
摘要： The spectral changes in surface-enhanced resonant Raman scattering (SERRS) and surface enhanced fluorescence (SEF) of single silver nanoparticle dimers adsorbed by near-single dye molecules are reproduced under strong coupling regimes. For the reproduction, the enhancement and quenching factors in SERRS and SEF are derived from the Purcell factors including both radiative and nonradiative plasmon modes. The Purcell factors are estimated using the coupling energies obtained by analyzing the spectral changes in plasmon resonance during SERRS and SEF decay processes on the basis of a classical hybridization model. The model is composed of a plasmon and a molecular exciton with phonon replicas accurately representing the molecular multi-level system. The reproduced SERRS spectral changes are consistent with the experimental ones. Furthermore, the calculated SEF spectral changes can reproduce the experimental ones by phenomenologically assuming transitions from ultra-fast SEF to conventional SEF with decreasing coupling energies.
关键词： strong coupling,hybridization model,silver nanoparticle dimer,dye molecules,Purcell factors,surface enhanced fluorescence,surface-enhanced resonant Raman scattering
更新于2025-09-23 15:23:52
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Determination of 17β-estradiol by surface-enhanced Raman spectroscopy merged with hybridization chain reaction amplification on Au@Ag core-shell nanoparticles
摘要： The authors describe an aptamer-based assay for 17β-estradiol. It relies on the combined use of surface enhanced Raman scattering (SERS) and hybridization chain reaction (HCR). The aptamer against 17β-estradiol is applied as the recognition probes, and this results in excellent specificity. Specific recognition of target 17β-estradiol induce the freedom of DNA 2, which will open the stem-loop structure of probe 1 on the Au@Ag and form the partial dsDNA structure. With the nicking enzyme, the partial dsDNA will be hydrolyzed and the reside ssDNA on Au@Ag will form a small stem-loop structure. With the help of the other probe 2 modified Au@Ag and pre-immobilized probe 3 on the well of the microplate, an enzyme-free HCR can occur and tremendous Au@Ag can be assembled along the formed dsDNA in HCR, which can act as the excellent substrate for Raman measurement and greatly amplify the Raman signal of R6G on the Au@Ag. Afterwards, the key factor, ratio between probe 2-Au@Ag (P2) and probe1-Au@Ag (P1), affects the detection sensitivity is systematically optimized for the best sensing performance. The SERS signal of R6G, best measured at 1651 cm?1, increases linearly in the wide range from 1 pM to 10 nM. The detection limit can be as low as 0.1 pM.
关键词： Estrogen,Hybridization chain reaction,SERS,Food safety,Aptamer,Gold nanoparticle,Signal amplification,Environment monitoring
更新于2025-09-23 15:23:52
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Graphene oxide-quenching-based fluorescence in situ hybridization (G-FISH) to detect RNA in tissue: Simple and fast tissue RNA diagnostics
摘要： FISH-based RNA detection in paraffin-embedded tissue can be challenging, with complicated procedures producing uncertain results and poor image quality. Here, we developed a robust RNA detection method based on graphene oxide (GO) quenching and recovery of fluorescence in situ hybridization (G-FISH) in formalin-fixed paraffin-embedded (FFPE) tissues. Using a fluorophore-labeled peptide nucleic acid (PNA) attached to GO, the endogenous long noncoding RNA BC1, the constitutive protein β-actin mRNA, and miR-124a and miR-21 could be detected in the cytoplasm of a normal mouse brain, primary cultured hippocampal neurons, an Alzheimer’s disease model mouse brain, and glioblastoma multiforme tumor tissues, respectively. Coding and non-coding RNAs, either long or short, could be detected in deparaffinized FFPE or frozen tissues, as well as in clear lipid-exchanged anatomically rigid imaging/immunostaining-compatible tissue hydrogel (CLARITY)-transparent brain tissues. The fluorescence recovered by G-FISH correlated highly with the amount of miR-21, as measured by quantitative real time RT-PCR. We propose G-FISH as a simple, fast, inexpensive, and sensitive method for RNA detection, with a very low background, which could be applied to a variety of research or diagnostic purposes.
关键词： glioblastoma multiforme tumor,tissue RNA diagnostics,Graphene oxide-quenching-based fluorescence in situ hybridization (G-FISH),Alzheimer’s disease,formalin-fixed paraffin-embedded (FFPE) tissue
更新于2025-09-23 15:23:52
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Preparation of fluorescent in situ hybridisation probes without the need for optimisation of fragmentation
摘要： DNA-fluorescence in situ hybridisation (DNA-FISH) allows visualisation of chromosome organisation and fluorescently labelled DNA fragments that are often produced from rearrangement. FISH probes are pools of short template plasmids that contain large genomic inserts. For effective sample penetration and target hybridisation it is critical that probe fragments are between 200 and 500bp. Production of these short probes requires significant optimisation and can be confounded access to expensive sonication equipment or inherent sequence features that influence enzymatic fragmentation or amplification. Here we demonstrate that effective FISH probes can be prepared without the need for optimisation of fragmentation using a cocktail of two the 4bp recognition sequence restriction enzymes CviQI and AluI.
关键词： Cancer,Fluorescence in situ hybridization,Translocation,FISH,Probe
更新于2025-09-23 15:23:52
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Peptide Nucleic Acid–Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study
摘要： PURPOSE. Rapid identi?cation of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-speci?c medication. The current study reports a new molecular application of peptide nucleic acid–?uorescence in situ hybridization (PNA-FISH) with Staphylococcus-speci?c molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis. METHODS. An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n ? 15), along with broth as negative controls (n ? 5) were used. Inoculum was prepared to a ?nal concentration of 1 3 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were ?xed and hybridized using QuickFISH protocol with probes for Staphylococcus. RESULTS. With PNA-FISH technique, Staphylococcus aureus was identi?ed in 9 of 10 samples and coagulase-negative staphylococci were identi?ed in 10 of 10 samples. Detection time was 20 minutes. CONCLUSIONS. This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identi?cation of isolates from patients with endophthalmitis.
关键词： peptide nucleic acid (PNA),endophthalmitis,rapid identification,fluorescence in situ hybridization (FISH)
更新于2025-09-23 15:22:29
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Signal-on Electrochemiluminescence Aptasensor for Bisphenol A based on Hybridization Chain Reaction and Electrically Heated Electrode
摘要： A simple and sensitive electrochemiluminescence (ECL) aptasensor has been developed for bisphenol A (BPA) detection. The capture DNA (CDNA) was modified on the heated indium-tin-oxide (ITO) working electrode surface firstly and then hybridized with BPA aptamer to form double strand DNA (dsDNA). The presence of target can cause the releasing of aptamer from the electrode surface since the aptamer prefers to switch its configuration to combine with BPA. Subsequently, the free CDNA will induce hybridization chain reaction (HCR) to produce long dsDNA on the electrode surface. Ru(phen)3 2+ can integrate into the grooves of dsDNA to act as an ECL reagent, thus enhanced ECL signal can be detected. The temperature control during the processes of target recognition and HCR were realized through the heated electrode instead of the bulk solution heating. Furthermore, the performance of the ECL aptasensor can be further enhanced at elevated electrode temperature. Under the optimized conditions, the ECL intensity of the system has a linear relationship with the logarithm of BPA concentration in the range of 2.0 pM-50 nM. The limit of detection (LOD) at 55 °C (electrode surface temperature) was calculated to be 1.5 pM, which was approximately 6.5-fold lower than that at 25 °C. The proposed biosensor has been applied to detect the BPA in drink samples with satisfactory results.
关键词： electrochemiluminescence,hybridization chain reaction,heated indium-tin-oxide electrode,aptamer,bisphenol A
更新于2025-09-23 15:22:29