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In this work, to enhance the fluorescence quantum yield of carbon dots (CDs), a novel metal-enhanced fluorescence (MEF) structure was designed by decorating CDs on silver nanoparticle (AgNPs) film. The glass slide-AgNPs (GS-AgNPs) structure was fabricated using the electrostatic adsorption method, and the AgNPs-CDs structures were prepared by the direct drying method, which then formed the GS-AgNPs-CDs composite structure. In this structure, the MEF effect was found to be size dependent by changing the 5 types of AgNPs with different sizes. And the MEF effect also decreased as the distance between the AgNPs and CDs increased by using polyvinylpyrrolidone (PVP) to separate the AgNPs and CDs. This hybrid structure can be used as a fluorescence detection platform and the recorded fluorescence intensity of GS-AgNPs 428 nm-CDs achieved a maximum enhancement factor (EF) of 31.72. Considering the high enhancement factor, this system may become promising to find potential applications in biochemical assay fields.

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Understanding driving forces for dissipative, i.e., out of equilibrium, assembly of nanoparticles from colloidal solution at liquid-solid interfaces provides the ability to design external cues for reconfigurable device response. Here electrohydrodynamic flow (EHD) at an electrode-liquid interface is investigated as a dissipative driving force for tuning optical response. EHD results from an oscillatory electric field in a liquid cell between two electrodes and drives assembly of gold nanoparticles (NP) into two-dimensional clusters on electrode surfaces. Clusters are chemically crosslinked during assembly to freeze assemblies for electron microscopy characterization in order to understand how to 'nucleate' cluster formation. Electron microscopy images show deposition with a potential having an amplitude of 5 V and frequency of 100 Hz produces surfaces with isolated NP, which can seed EHD flow. A second deposition step at 5 V and 500 Hz produces a high density of quadramers on surfaces. When exciting near the local surface plasmon resonance of the Au NP clusters formed during assembly, Au NPs serve as in situ nanoantenna reporters of assembly and disassembly. Surface enhanced Raman scattering (SERS) measurements of Au NP capped with 4-mercaptobenzoic acid show order of magnitude signal enhancements occur during cluster formation in the presence of an oscillatory field, which occurs on a time scale of seconds. Confocal fluorescence spectroscopy is used to monitor the dissipative assembly of Au NP over multiple cycles. Results provide insight on how electrical stimuli and seeding local perturbations affects formation of NP clusters and resultant optical response provides insight on how to tune response of optically active surfaces.

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Point-of-care testing (POCT) for low-concentration protein biomarkers remains challenging due to limitations in biosensor sensitivity and platform integration. This study addresses this gap by presenting a novel approach that integrates a metal-enhanced fluorescence (MEF) biosensor within a capillary flow-driven microfluidic cartridge (CFMC) for the ultrasensitive detection of the Parkinson's disease biomarker, aminoacyl-tRNA synthetase complex interacting multi-functional protein 2 (AIMP-2). Crucial point to this approach is the orientation-controlled immobilization of capture antibody on a nanodimple-structured MEF substrate within the CFMC. This strategy significantly enhances fluorescence signals without quenching, enabling accurate quantification of low-concentration AIMP-2 using a simple digital fluorescence microscope with a light-emitting diode excitation source and a digital camera. The resulting platform exhibits exceptional sensitivity, achieving a limit of detection in the pg/mL range for AIMP-2 in human serum. Additionally, the CFMC design incorporates a capillary-driven passive sample transport mechanism, eliminating the need for external pumps and further simplifying the detection process. Overall, this work demonstrates the successful integration of MEF biosensing with capillary microfluidics for point-of-care applications.

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Matrix metalloproteinases (MMPs) are attractive biomarkers for cancer diagnosis and treatment, while it is still a challenge to precise analysis of MMP activities owing to their very low abundance in the biological samples, especially at the early stages of tumors. Herein, a peptide microarray-based metal-enhanced fluorescence assay (PMMEFA) is proposed to simultaneously detect MMP-1, -2, -3, -7, -9, and -13 activities. The assay involves immobilization of Förster resonance energy transfer dye pair decorated peptides (FRET-peptides) on a poly(glycidyl methacrylate-co-2-hydroxyethyl methacrylate) coated gold nanorod modified glass slide (GNR@P(GMA-HEMA)). To fabricate the GNR@P(GMA-HEMA) slide, GNRs are self-assembled onto an aminated glass slide, and a polymer brush (P(GMA-HEMA)) is grown through a surface-initiated atom transfer radical polymerization reaction (SI-ATRP). Upon the addition of MMPs, the FRET pairs are broken due to the specific cleavage of FRET-peptides by enzymes, resulting in the recovery of fluorescence signals and further enhancement by the MEF of GNRs. The fluorescence recovery degree provides a direct indicator for MMP activity. The PMMEFA exhibits excellent sensitivity, which enables to detect MMP-1, -2, -3, -7, -9, and -13 activities, with low limits of detection (LODs) of 1.7 fg mL-1, 0.3 fg mL-1, 2.0 fg mL-1, 1.8 fg mL-1, 2.2 fg mL-1 and 14.0 fg mL-1, respectively. To substantiate the practicability of PMMEFA, MMP activities were measured in a range of matrices, encompassing cell culture medium, serum, and tumor tissue homogenate, and MMP activities can be detected only in 0.15 µL serum and 0.025 mg tumor tissue.

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The implementation of metal enhanced fluorescence (MEF) as an efficient detection tool, especially in the near-infrared region of the electromagnetic spectrum, is a rather new direction for diagnostic analytical technologies. In this context, we propose a novel microfluidic plasmonic design based on paper for efficient MEF detection of the "proof-of-concept" biotin-streptavidin recognition interaction. Our design made use of the benefits of gold nanobipyramids (AuBPs), considering the strong enhanced electromagnetic field present at their sharp tips, and filter paper to operate as a natural microfluidic channel due to excellent wicking abilities. The calligraphed plasmonic paper, obtained using a commercial pen filled with AuBPs, was integrated in a robust sandwich optically transparent polydimethylsiloxane chip, exhibiting portability and flexibility while preserving the chip's properties. To place the Alexa 680 fluorophore at an optimal distance from the nanobipyramid substrate, the human IgG-anti-IgG-conjugated biotin sandwich reaction was employed. Thus, upon the capture of Alexa 680-conjugated streptavidin by the biotinylated system, a 1.3-fold average enhancement of the fluorophore's emission was determined by bulk fluorescence measurements. However, the local enhancement factor was considerably higher with values spanning from 5 to 6.3, as proven by mapping the fluorescence emission under both re-scan microscopy and fluorescence lifetime imaging, endorsing the proposed chip's feasibility for bulk MEF biosensing as well as high-resolution MEF bioimaging. Finally, the versatility of our chip was demonstrated by adapting the biosensing protocol for cardiac troponin I biomarker detection, validated using 10 plasma samples collected from pediatric patients and corroborated with a conventional ELISA assay.

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In this communication luminescent bioconjugated human serum albumin nanostructures (HSA NPs) with tiny ultraluminescent gold core-shell silica nanoparticles (Au@SiO2-Fl) were designed with enhanced bi-coloured luminescence properties. The HSA NPs were obtained from Human Serum Albumin free (HSA free) through the desolvation method, and Au@SiO2-Fl, through modified Turkevich and Störber methods. In this manner, porous HSA Nanostructures of 150.0-200 nm and Au@SiO2-Fl 45.0 nm final diameters were obtained. Both methodologies and structures were conjugated to obtain modified Nanocomposites based on tiny gold cores of 15 nm surrounded with well spatial Nanostructured architectures of HSA (d15 Au@SiO2-Fl-HSA) that generated variable nanopatterns depending on the modified methodology of synthesis applied within colloidal dispersions. Therefore, three methodologies of non-covalent conjugation were developed. In optimal conditions, through Transmission Electronic Microscopy (TEM), well resolved multilayered nano-architectures with a size 190.0-200 nm in average with variable contrast depending of the focused nanomaterial within the nanocomposite were shown. Optimized nanoarchitecture was based on a template tiny gold core-shell surrounded by nanostructured HSA NPs (d15 Au@SiO2-Fl-HSA). In this manner, the NanoImaging generated by laser fluorescence microscopy permitted to record improved optical properties and functionalities, such as: (i) enhanced ultraluminescent d15 Au@SiO2-Fl-HSA composites in comparison to individual components based on Metal Enhanced Fluorescence (MEF); (ii) diminished photobleaching; (iii) higher dispersibility; (iv) higher resolution of single bright nano-emitters of 210.0 nm sizes; and (v) enhanced bi-coloured Bio-MEF coupling with potential non-classical light delivery towards other non-optical active biostructures for varied applications. The characterization of these nanocomposites allowed the comparison, evaluation and discussion focused on new properties generated and functionalities based on the incorporation of different types of tuneable materials. In this context, the biocompatibility, Cargo confined spaces, protein-based materials, optical transparent could be highlighted, as well as optical active materials. Thus, the potential applications of nanotechnology to both nanomedicine and nano-pharmaceutics were discussed.

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When exposed to specific light wavelengths, carbon dots (CDs), which tend to be fluorescent, can emit colorful light. It provides them with a lot of adaptability for different applications including bioimaging, optoelectronics, and even environmental sensing. Poly(ethylenimine) (PEI) coated carbon dots (PEI-CDs) with a long emission wavelength were synthesized via the hydrothermal method. The resultant CDs show strong fluorescence with quantum yield up to 20.2%. The PEI-CDs exist with distinct pH-sensitive features with pH values in the range of 2-14. The optical characteristics of CDs are pH-responsive due to the presence of different amine groups on PEI, which is a functional polycationic polymer. One of the most widely employed nanoparticles for improving the fluorescence plasmonic characteristics of a nanocomposite is gold. Gold nanoparticles were coupled with PEI-CDs in this assay by using the EDC-NHS coupling to increase the photoluminescence property of the PEI-CDs by using the metal-enhanced fluorescence approach. In the presence of gold nanoparticles, the fluorescence is enhanced 5-6 times. The likely mechanism in our investigation was primarily derived from enhancement of the intrinsic radiative decay rate rather than the local electric field impact. Moreover, PEI-CDs can be used as a bioimaging agent, as these molecules are nontoxic to the cells, and the positively charged PEI-CDs have the potential for nuclear targeting, allowing for electrostatic contact with DNA in the nucleus. This finding will expand the application that the PEI-CDs can be used in the future for targeted imaging applications.

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Metal-enhanced photoluminescence is able to provide a robust signal even from a single emitter and is promising in applications in biosensors and optoelectronic devices. However, its realization with semiconductor nanocrystals (e.g., quantum dots, QDs) is not always straightforward due to the hidden and not fully described interactions between plasmonic nanoparticles and an emitter. Here, we demonstrate nonclassical enhancement (i.e., not a conventional electromagnetic mechanism) of the QD photoluminescence at nonplasmonic conditions and correlate it with the charge exchange processes in the system, particularly with high efficiency of the hot-hole generation in gold nanoparticles and the possibility of their transfer to QDs. The hole injection returns a QD from a charged nonemitting state caused by hole trapping by surface and/or interfacial traps into an uncharged emitting state, which leads to an increased photoluminescence intensity. These results open new insights into metal-enhanced photoluminescence, showing the importance of the QD surface states in this process.

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Rapid diagnostic tests based on the lateral flow immunoassay (LFI) enable early identification of viral infection, owing to simple interpretation, short turnaround time, and timely isolation of patients to minimize viral transmission among communities. However, the LFI system requires improvement in the detection sensitivity to match the accuracy of nucleic acid amplification tests. Fluorescence-based LFIs are more sensitive and specific than absorption-based LFIs, but their performance is significantly affected by fundamental issues related to the quantum yield and photobleaching of fluorophores. Metal-enhanced fluorescence (MEF), which is a plasmonic effect in the vicinity of metallic nanoparticles, can be an effective strategy to improve the detection sensitivity of fluorescence-based LFIs. The key factors for obtaining a strong plasmonic effect include the distance and spectral overlap of the metal and fluorophore in the MEF system. In this study, MEF probes were designed based on core-shell nanostructures employing a gold nanorod core, mesoporous silica shell, and cyanine 5 fluorophore. To optimize the efficiency of MEF probes incorporated on the LFI platform (MEF-LFI), we experimentally and theoretically investigated the distance dependence of plasmonic coupling between cyanine 5 and gold nanorods by adjusting the shell thickness, resulting in significant fluorescence enhancement. The proposed MEF-LFI enabled highly sensitive detection of influenza A virus (IAV) nucleocapsid protein with a detection limit of 0.52 pg mL-1 within 20 min and showed high specificity and accuracy for determining IAV clinical samples. Overall, our findings demonstrate the potential of this method as an effective tool for molecular diagnosis under emergency conditions.

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In this study, a facile one-pot solid-state synthesis method is developed to shed light on the metal-enhanced fluorescence (MEF) effect in carbon quantum dots (CQDs) and gold nanoparticles (AuNPs) hybrid materials. This is one of the few studies on the solid-state synthesis of N-doped CQDs/gold hybrid nanomaterials. We have conducted various sets of experiments to reveal the role of individual reagents during the nucleation and growth of nanoparticles. We have demonstrated that the addition of a small amount of gold salt illustrates a paramount effect (103-fold) in photoluminescence intensity. This effect is ascribed to MEF, which is caused due to interactions between the excited-state fluorophores and the free surface electrons of metal nanoparticles. It is interesting to note that a further increase of gold yields fluorescence quenching due to a large number of formed AuNPs causing fluorescence resonance energy transfer. By adjusting the volume ratio of gold salt and CD precursors, it is possible to obtain the CQDs-AuNPs hybrid with the highest fluorescence, which produces extensive visible light under 460 nm excitation. Synthesized materials have been successfully used for imaging human dermal fibroblasts and A549 lung epithelial cells. The dose-dependent cytotoxicity studies reveal that the hybrid structures do not have cytotoxicity.

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The purpose of this paper is to provide an in-depth review of plasmonic metal nanoparticles made from rhodium, platinum, gold, or silver. We describe fundamental concepts, synthesis methods, and optical sensing applications of these nanoparticles. Plasmonic metal nanoparticles have received a lot of interest due to various applications, such as optical sensors, single-molecule detection, single-cell detection, pathogen detection, environmental contaminant monitoring, cancer diagnostics, biomedicine, and food and health safety monitoring. They provide a promising platform for highly sensitive detection of various analytes. Due to strongly localized optical fields in the hot-spot region near metal nanoparticles, they have the potential for plasmon-enhanced optical sensing applications, including metal-enhanced fluorescence (MEF), surface-enhanced Raman scattering (SERS), and biomedical imaging. We explain the plasmonic enhancement through electromagnetic theory and confirm it with finite-difference time-domain numerical simulations. Moreover, we examine how the localized surface plasmon resonance effects of gold and silver nanoparticles have been utilized for the detection and biosensing of various analytes. Specifically, we discuss the syntheses and applications of rhodium and platinum nanoparticles for the UV plasmonics such as UV-MEF and UV-SERS. Finally, we provide an overview of chemical, physical, and green methods for synthesizing these nanoparticles. We hope that this paper will promote further interest in the optical sensing applications of plasmonic metal nanoparticles in the UV and visible ranges.

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A competitive fluorescent immunoassay is described for the ultrasensitive determination of amyloid beta peptide1-42 (Aß1-42), a biomarker for early diagnosis of Alzheimer's disease. N, S-doped graphene quantum dots (N, S-GQDs) were freely assembled on the surface of Ag@SiO2 nanoparticles to obtain a composite (Ag@SiO2@N, S-GQD nanocomposite), which was successfully prepared and characterized. By theoretical study, the optical properties of nanocomposites are improved compared with GQDs, due to the advantages of combining N, S co-doping and metal-enhanced fluorescence (MEF) effect of Ag NPs. In addition, Aß1-42 was modified by Ag@SiO2@N, S-GQDs to prepare a probe with high photoluminescence properties (Ag@SiO2@N, S-GQDs-Aß1-42). In the presence of Aß1-42, a competitive reaction towards anti-Aß1-42 fixed on the ELISA plate was proceeded between Aß1-42 and Ag@SiO2@N, S-GQDs-Aß1-42 by specific capture of antigen-antibody. The emission peak of Ag@SiO2@N, S-GQDs-Aß1-42 (400 nm emission) was used for the quantitative determination of Aß1-42. Under the optimal conditions, the fluorescent immunoassay exhibited a linear range of 0.32 pg·mL-1-5 ng·mL-1 with a detection limit of 0.098 pg·mL-1. The results show that the immunoassay has good analytical ability and can provide a new method for the clinical determination of Aß1-42.

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Metal-enhanced fluorescence (MEF) is an important fluorescence technology due to its ability to significantly improve the fluorescence intensity. Here, we present a new MEF configuration of the bionic nanorod array illuminated by radially polarized vector beam (RVB). The bionic nanorod array is fabricated via a nanoimprinting method by using the wings of the Chinese cicada "meimuna mongolica" as bio-templates, and later coating gold film by ion sputtering deposition method. The MEF performance of the prepared substrate is tested by a home-made optical system. The experiment results show that, in the case of RVB excitation, the intensity of fluorescence is more than 10 times stronger with the nano-imprinted substrate than that with glass. Using the bionic nanoarray as a substrate, the intensity of fluorescence is ~2 times stronger via RVB than that by the linearly polarized beam. In addition, the prepared substrate is verified to have good uniformity.

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Conventional cellular protein detection techniques such as immunocytochemistry and flow cytometry require abundant cells, posing multiple challenges, including difficulty and cost for obtaining enough cells and the potential for clogging the instrument when using flow cytometry. Also, it is challenging to conduct cellular protein imaging and quantification simultaneously from a single experiment. We present a novel 3D platform, which integrates highly biocompatible cell-entrapped alginate hydrogel droplet array with gold-nanoparticle (AuNP)-based metal enhanced fluorescence (MEF), to achieve simultaneous imaging and quantification of proteins in intact cells in a sensitive manner. Compared to 2D immunocytochemistry, this 3D system allows for a higher cell loading capacity per unit area; together with the MEF-based signal enhancement from the embedded AuNPs, sensitive protein quantification was realized. Furthermore, compared to flow cytometry, this platform allows for protein imaging from individual cells. Taking the detection of EpCAM protein in ovarian cancer cells as a model, we optimized the AuNP size and concentration for optimal fluorescent signals. The 5 nm AuNPs at 6.54 × 1013 particles/mL proved to be the most effective in signal enhancement, providing 2.4-fold higher signals compared to that without AuNPs and 6.4-fold higher signals than that of 2D immunocytochemistry. The number of cells required in our technology is 1-3 orders of magnitude smaller than that of conventional methods. This AuNP-embedded hydrogel platform combines the benefits of immunocytochemistry and flow cytometry, providing increased assay sensitivity while also allowing for qualitative analysis through imaging, suitable for protein determination in a variety of cells.

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Fluorescence can be enhanced or quenched depending on the distance between the surface of a metal nanoparticle and the fluorophore molecule. Fluorescence enhancement by nearby metal particles is called metal-enhanced fluorescence (MEF). MEF shows promising potential in the field of fluorescence-based biological sensing. MEF-based biosensor systems generally fall into two platform categories: (1) a two/three-dimensional scaffold, or (2) a colloidal suspension. This review briefly summarizes the application studies using wavelength-dependent carbon dots (UV-VIS), noble metals (VIS), and upconversion nanoparticles (NIR to VIS), representative nanomaterials that contribute to the enhancement of fluorescence through the resonance energy transfer modulation and then presents a perspective on this topic.

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Fluorescence-based biosensors have widely been used in the life-sciences and biomedical applications due to their low limit of detection and a diverse selection of fluorophores that enable simultaneous measurements of multiple biomarkers. Recent research effort has been made to implement fluorescent biosensors into the exploding field of point-of-care testing (POCT), which uses cost-effective strategies for rapid and affordable diagnostic testing. However, fluorescence-based assays often suffer from their feeble signal at low analyte concentrations, which often requires sophisticated, costly, and bulky instrumentation to maintain high detection sensitivity. Metal- and metal oxide-based nanostructures offer a simple solution to increase the output signal from fluorescent biosensors due to the generation of high field enhancements close to a metal or metal oxide surface, which has been shown to improve the excitation rate, quantum yield, photostability, and radiation pattern of fluorophores. This article provides an overview of existing biosensors that employ various strategies for fluorescence enhancement via nanostructures and have demonstrated the potential for use as POCT. Biosensors using nanostructures such as planar substrates, freestanding nanoparticles, and metal-dielectric-metal nanocavities are discussed with an emphasis placed on technologies that have shown promise towards POCT applications without the need for centralized laboratories.

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To improve the direct quantification of Carcinoembryonic Antigen (CEA) from body fluids by immunofluorescence, a surface acoustic wave (SAW) based biosensor was developed combined with an optimized silver nanostructure at the sensing region. Fluorescence signal amplification is achieved by patterning silver nanostructures using the rapid thermal annealing (RTA) method. In addition, the problem of background noise interference from nonspecific binding in human plasma is addressed by Rayleigh wave streaming at the immunoassay region, which shows a reduction in the limit of detection. The results show that the silver nanostructures significantly increase the sensor sensitivity by 49.99-fold and lower the limit of detection of CEA in phosphate buffered saline (PBS) solution to 101.94 pg/mL. The limit of detection of CEA biomarker in human plasma was successfully brought down to 11.81 ng/mL by reducing background noise using Rayleigh SAW streaming. This allows for a point-of-need sensor system to be realized in various clinical biosensing applications.

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Plasmonic micro/nanobeads exhibit unique physicochemical properties attributed to their localized surface plasmon resonance (LSPR), enabling use in sensitive suspension array assays and matrix-assisted laser deposition/ionization mass spectrometry (MALDI-MS) analysis. Herein, we report a facile method for the preparation of magnetic plasmonic micro/nanobeads by the combination of Shirasu porous glass (SPG) membrane emulsification and polydopamine (PDA)-assisted in-situ reduction. The magnetic responsiveness properties endowed by doped Fe3O4 nanoparticles result in easy and complete separation of unwanted components during the preparation and bio-reaction processes. In addition, the coverage degree of the plasmonic shell can be flexibly controlled. As a result of the significant metal-enhanced fluorescence effect, as-prepared plasmonic microbeads enable the sensitive detection of alpha-fetoprotein (AFP) and deoxyribonucleotide (DNA) in suspension array with detection limits of 0.11 ng mL-1 and 1.65 fmol mL-1, respectively, 8.6 times and 2 orders of magnitude higher than unmodified microbeads. Furthermore, as-prepared plasmonic nanobeads can be used as a matrix for MALDI-MS to allow the detection of low molecular weight biological molecules. As little as 0.2 pmol of proline and serine can be detected in a sample as small as 0.5 µL. This work provides a general strategy for the design of multifunctional plasmonic micro/nanomaterials that will help promote further advancements in sample analysis.

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Dye-doped nanoparticles have been investigated as bright, fluorescent probes for localization-based super-resolution microscopy. Nanoparticle size is important in super-resolution microscopy to get an accurate size of the object of interest from image analysis. Due to their self-blinking behavior and metal-enhanced fluorescence (MEF), Ag@SiO2 and Au@Ag@SiO2 nanoparticles have shown promise as probes for localization-based super-resolution microscopy. Here, several noble metal-based dye-doped core-shell nanoparticles have been investigated as self-blinking nanomaterial probes. It was observed that both the gold- and silver-plated nanoparticle cores exhibit weak luminescence under certain conditions due to the surface plasmon resonance bands produced by each metal, and the gold cores exhibit blinking behavior which enhances the blinking and fluorescence of the dye-doped nanoparticle. However, the silver-plated nanoparticle cores, while weakly luminescent, did not exhibit any blinking; the dye-doped nanoparticle exhibited the same behavior as the core fluorescent, but did not blink. Because of the blinking behavior, stochastic optical reconstruction microscopy (STORM) super-resolution analysis was able to be performed with performed on the gold core nanoparticles. A preliminary study on the use of these nanoparticles for localization-based super-resolution showed that these nanoparticles are suitable for use in STORM super resolution. Resolution enhancement was two times better than the diffraction limited images, with core sizes reduced to 15 nm using the hybrid Au-Ag cores.

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Exosomes, carrying specific molecular information of their parent cells, have been regarded as a kind of promising noninvasive biomarker for liquid biopsy. Plentiful fluorescence methods have been proposed for exosome assay. However, most of them are dependent on nucleic acid signal amplification strategies, which require complicated sequence design and experimental operation. Herein, a metal-enhanced fluorescence (MEF) biochip based on shell-isolated Au@MnO2 nanoparticle array was designed for simple and sensitive assay of exosomes. The designed method consists of only two parts: signal conversion and MEF amplification. The conversion of exosome signals to DNA signals was realized by means of chain displacement reaction. The subtle conversion effectively averts the effect of steric hindrance on MEF while amplifying the signal easily for the first time. The MEF biochip based on shell-isolated Au@MnO2 nanoparticle array achieves a second signal amplification in a simple way. Profiting from the two signal amplifications, this strategy displays high sensitivity toward exosomes with a detection limit of 4.5 × 103 particles µL-1. Compared with the result without MEF, the sensitivity is enhanced about thirty times. As far as we know, this is the first attempt for exosome assay by using MEF strategy. In addition to the favorable fluorescence enhancement, both shell-isolated Au@MnO2 nanoparticles and Au@MnO2 nanoparticle array show excellent stability in buffer solutions, which is conducive to practical application. Moreover, the proposed method is able to distinguish breast cancer patients from healthy people, showing its potential for exosome-based liquid biopsy.

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