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Reactive oxygen species (ROS) are key regulators in numerous pathological contexts, including cancer or inflammation. Their role is complex, which justifies the need for methods enabling their quantitative and time-resolved monitoring in vivo, in the perspective to profile tissues of individual patients. However, current ROS detection methods do not provide these features. Here, we propose a new method based on the imaging of lanthanide-ion nanoparticles (GdVO4:Eu), whose photoluminescence is modulated by the surrounding ROS concentration. We monitored their luminescence after intradermic injection in a mouse ear submitted to an inflammation-inducing topical stimulus. Based on this approach, we quantified the ROS concentration after inflammation induction and identified a two-step kinetics of ROS production, which may be attributed to the response of resident immune cells and their further recruitment at the inflammation locus.
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Lateral Flow Assays (LFAs) have been extensively used on-site to rapidly detect analytes, possibly in complex media. However, standard gold nanoparticle-based LFAs lack sensitivity and cannot provide quantitative measurements with high accuracy. To overcome these limitations, we image lanthanide-doped nanoparticles (YVO4:Eu 40%) as new luminescent LFA probes, using a homemade reader coupled to a smartphone and propose an original image analysis allowing strip quantification regardless of the shape of the test band signal. This method is demonstrated for the detection of staphylococcal enterotoxins SEA, SEG, SEH, and SEI. A systematic comparison to state-of-the-art gold nanoparticle-based LFA revealed an analytical sensitivity enhancement of at least one order of magnitude. We furthermore provided measurements of absolute toxin concentration over two orders of magnitude and demonstrated simultaneous quantitative detection of multiple toxins with unaltered sensitivity. In particular, we reached concentrations 100 times lower than the ones reported in the literature for on-site multiplexed LFA targeting enterotoxins. Altogether, these results highlight that our luminescent nanoparticle-based method provides a powerful and versatile on-site framework to detect multiple biomolecules with sensitivity approaching that obtained by ELISA. This paves the way to a change of paradigm in the field of analytical immunoassays by providing fast in situ quantitative high sensitivity detection of biomarkers or pathogens.
Asunto(s)
Elementos de la Serie de los Lantanoides , Nanopartículas del Metal , Enterotoxinas , Oro , InmunoensayoRESUMEN
Trapping of NaYF4:Er/Yb/Gd nanorods using an original optical fiber-tip tweezers is reported. Depending on their length, nanorods are reproducibly trapped in single or dual fiber tip configurations. Short rods of 600 nm length are trapped with two fiber tips facing each other. In contrary, long rods (1.9 µm) can be stably trapped at the apex of one single fiber tip and at a second stable trapping position 5 µm away from the tip. The up-conversion emission of trapped long nanorods is studied as a function of the position on the nanorod and in three orthogonal directions. The experimental results are discussed using numerical simulations based on exact Maxwell Stress Tensor approach.
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Metal dielectric nanocomposites exhibit a broad range of physical properties that can be tuned by varying the metallic volume fraction (Ï), in particular near the percolation threshold. The study and exploitation of the so-called critical properties at this threshold are currently limited by the inability to finely tune Ï in a continuous way, for example in physical mixtures. Here we present a novel chemical fabrication process for metal dielectric coatings consisting of TiO2 nanoparticles dispersed in a mesoporous silver silicate host matrix. UV light irradiation of these active films allows for the photocatalytic formation of silver nanoparticles with a dose dependent, measured incremental variation in Ï as small as 0.01% up to a total volume fraction of 20% (i.e. exceeding the 3D percolation threshold). Moreover, this is achieved in situ, that is to say, while measuring the optical or electrical properties of the nanocomposite. Simple examples of hysteretic resistance measurements and optical absorption/reflection as a function of Ï are presented, demonstrating the utility of this nanocomposite system for the study of percolation phenomena as well as its potential for applications such as sensitive strain sensors.
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Reactive oxygen species (ROS), and notably hydrogen peroxide H2O2, are cellular second messengers that are known to control a variety of signaling processes. They can finely regulate the dynamics of signal transduction, cell response and ultimately tissue function. However, there are very few local, quantitative and time-resolved descriptions of their cellular organization at the scale of molecular reactions, due to the lack of efficient sensors. We thus developed a novel nanoprobe-based ROS detection system using the simultaneous imaging of single lanthanide nanoparticles (YAG:Ce and chemically reduced Gd0.6Eu0.4VO4). We reveal that both particle luminescence signals are controlled by their H2O2 local environment. By simultaneously tracking their luminescence, we devised a new approach providing a quantitative (0.5 µM accuracy in the 1-10 µM range) H2O2 measurement with a 500 ms time resolution, surpassing all existing methods by two orders of magnitude, and revealing previously inaccessible molecular events controlling ROS concentration. We used this nanoprobe in living cells to track fast signaling pathways, by measuring the dynamics of H2O2 intracellular concentrations, induced by endothelin-1 (ET-1) stimulation. We thus revealed the mechanisms controlling ROS production, notably the activity modulation of the ROS-producing enzyme NADPH oxidase by fast (<10 s) EGFR transactivation, and measured quantitatively their kinetic parameters through a minimal analytical model. Altogether, these results illustrate how lanthanide nanoparticle-based sensors are a powerful tool to dynamically probe molecular mechanisms shaping the oxidative cell response.
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Elementos de la Serie de los Lantanoides/química , Metales de Tierras Raras/química , Miocitos del Músculo Liso/metabolismo , Nanopartículas , Especies Reactivas de Oxígeno/metabolismo , Transducción de Señal , Animales , Endotelina-1/metabolismo , Peróxido de Hidrógeno , Cinética , Ratones Endogámicos C57BL , Músculo Liso Vascular/citología , Cultivo Primario de CélulasRESUMEN
The goal of this study is to elaborate few-micrometer thick optically active coatings based on nanoparticles spray-deposited onto a substrate and to control their scattering properties through a progressive suppression of the coffee-ring effect. The modification of the aggregation state of the nanoparticles to be sprayed induces a change of the surface roughness of the films and consequently of their optical transmission. We draw the counterintuitive conclusion that a nonstable colloidal solution gives a smoother coating than a highly stabilized colloidal solution, leading to a more transparent coating. This phenomenon is demonstrated in the case of commercial TiO(2) nanoparticles, as well as of homemade luminescent YVO(4):Eu nanoparticles, and seems to be generalized to a large range of systems.
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Hydrogen terminations (C-H) confer to diamond layers specific surface properties such as a negative electron affinity and a superficial conductive layer, opening the way to specific functionalization routes. For example, efficient covalent bonding of diazonium salts or of alkene moieties can be performed on hydrogenated diamond thin films, owing to electronic exchanges at the interface. Here, we report on the chemical reactivity of fully hydrogenated High Pressure High Temperature (HPHT) nanodiamonds (H-NDs) towards such grafting, with respect to the reactivity of as-received NDs. Chemical characterizations such as FTIR, XPS analysis and Zeta potential measurements reveal a clear selectivity of such couplings on H-NDs, suggesting that C-H related surface properties remain dominant even on particles at the nanoscale. These results on hydrogenated NDs open up the route to a broad range of new functionalizations for innovative NDs applications development.
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Nanodiamantes/química , Alquenos/química , Compuestos Azo/química , Carbono/química , Hidrógeno/química , Hidrogenación , Espectroscopía de Fotoelectrones , Presión , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , TemperaturaRESUMEN
We present a simple method based on the dispersion of fluorescent quantum dots (QD) into a liquid crystal phase that provides either nanostructured material or isolated QD micelles depending on water concentration. The liquid-crystal phase was obtained by using a gallate amphiphile with a poly(ethylene glycol) chain as the polar headgroup, named I. The hydration of QD/I mixtures resulted in the formation of a composite hexagonal phase identified by small-angle X-ray scattering and by polarized light and fluorescence optical microscopy, showing a homogeneous distribution of fluorescence within hexagonal phase. This composite mesophase can be converted into isolated QD-I micelles by dilution in water. The fluorescent QD-I micelles, purified by size exclusion chromatography, are well monodisperse with a hydrodynamic diameter of 20-30 nm. Moreover, these QD do not show any nonspecific adsorption on lipid or cell membranes. By simply adjusting the water content, the PEG gallate amphiphile I provides a simple method to prepare a self-organized composite phase or pegylated water soluble QD micelles for biological applications.