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The over- and under-expression of certain proteins in extracellular vesicles has been observed in many physiological and pathological conditions; however, a simple method to sort vesicles based on contrast in protein content is yet to be developed. We herein present a nonaffinity-based method for rapid and inexpensive isolation of lipid vesicles based on their membrane protein content. Based on a composition-specific thermophysical property change of vesicles at different protein contents, an acoustic property change that enabled an acoustophoretic separation was observed. This change was demonstrated in a thermally modulated acoustofluidic device in the form of a shift in vesicle migration from the nodal plane to antinodal plane at a specific temperature known as the acoustic contrast temperature (TΦ). Using phosphatidylcholine vesicles containing the membrane proteins gramicidin D, alamethicin, and melittin at molar contents ranging from 0.001% to 10%, we observed that increasing the membrane protein content brought about conformational changes in the membrane which afforded the vesicles distinctive acoustic properties. Then, by establishing an acoustic contrast temperature window, vesicles with the same protein but different molar content were successfully separated. The efficiency of the separation was studied for various vesicle mixtures and a separation efficiency as high as 97% was accomplished. In order to confirm the technique's applicability for biological samples, sheep red blood cells with various melittin peptide contents similarly demonstrated the depressing effects of melittin on membrane bending modulus and depressed the TΦ of the cells. This method holds promise for a myriad of applications in the biomedical field, especially in bioanalytical research.
Assuntos
Acústica , Separação Celular , Proteínas de Membrana/química , Técnicas Analíticas Microfluídicas , Temperatura , Acústica/instrumentação , Alameticina/análise , Animais , Separação Celular/instrumentação , Eritrócitos/química , Gramicidina/análise , Lipídeos/química , Meliteno/análise , Técnicas Analíticas Microfluídicas/instrumentação , Estrutura Molecular , Tamanho da Partícula , Ovinos , Propriedades de SuperfícieRESUMO
Gel polymer electrolytes (GPE) and composite GPE (cGPE) using one-dimensional glass microfillers have been developed for their use in lithium-oxygen batteries. Using glass microfillers, tetraglyme solvent, UV-curable polymer, and lithium salt at various concentrations, the preparation of cGPE yielded free-standing films. These cGPEs, with 1 wt % of microfillers, demonstrated increased ionic conductivity and lithium transference number over GPEs at various concentrations of lithium salt. Improvements as high as 50% and 28% in lithium transference number were observed for 0.1 and 1.0 mol kg-1 salt concentrations, respectively. Lithium-oxygen batteries containing cGPE similarly showed superior charge/discharge cycling for 500 mAh g-1 cycle capacity with as high as 86% and 400% increase in cycles for cGPE with 1.0 and 0.1 mol kg-1 over GPE. Results using electrochemical impedance spectroscopy, Raman spectroscopy, and scanning electron microscopy revealed that the source of the improvement was the reduction of the rate of lithium carbonates formation on the surface of the cathode. This reduction in formation rate afforded by cGPE-containing batteries was possible due to the reduction of the rate of electrolyte decomposition. The increase in solvated to paired Li+ ratio at the cathode, afforded by increased lithium transference number, helped reduce the probability of superoxide radicals reacting with the tetraglyme solvent. This stabilization during cycling helped prolong the cycling life of the batteries.
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Biomechanical properties of cells such as cellular stiffness have been increasingly considered as biomarkers for diseases. For instance, stiffness of cancer cells has been correlated to the malignant potential in certain cell lines. In cells, the cholesterol content plays a crucial role in determining stiffness. Changes in the cholesterol content in cellular membranes can be an indication of pathological disorders. Acoustophoresis as a separation and diagnostic tool is well positioned to help in the separation and diagnosis of cells taking advantage of its unique separation criteria of density and compressibility. However, under the same conditions, cells and vesicles secreted by these cells often have a positive contrast factor sign and thus do not yield simple separations. Thermally-assisted acoustophoresis, also referred to as thermo-acoustophoresis, solves this problem by adding a temperature dimension to the separation. In this work, we evaluate the acoustic contrast temperature (TΦ) of vesicles at different cholesterol molar ratios (Xchol) and develop a multi-stage lab-on-a-chip method to accomplish for the first time the separation of a three-vesicle mixture. Using Xchol = 0.1, 0.2, and 0.3 vesicles, we have obtained separation efficiencies exceeding 93%. The simplicity, rapidity, and label-free nature of this approach holds promise as a diagnostic and separation tool for cells and extracellular vesicles such as exosomes and microvesicles.
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Acústica/instrumentação , Colesterol/química , Vesículas Citoplasmáticas/química , Dispositivos Lab-On-A-Chip , Colesterol/análise , Modelos Biológicos , Fosfatidilcolinas/química , TemperaturaRESUMO
We report on a newly-developed membrane stiffness-based separation of vesicles using a thermally-assisted acoustophoretic approach. By tuning the temperature, we achieved the separation of vesicles of the same size, shape, and charge but with different stiffness values. It was observed that at a specific transition point, the acoustic contrast factor of vesicles changed sign from positive to negative. This change was mainly due to the change in the acoustic compressibility of the vesicles, which is inversely proportional to stiffness. The acoustic contrast temperature, corresponding to the temperature at which the acoustic contrast factor switches sign, was determined to be unique to the composition of the vesicles. This unique temperature signature allowed us to develop a separation method of vesicles with distinct membrane stiffness with target outlet purities exceeding 95%. Our studies suggest that this method may be applied for the separation of cells affected by diseases that affect the cellular stiffness.
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Temperatura , Ondas Ultrassônicas , Lipossomas UnilamelaresRESUMO
Over the past several years, nanomaterials derived from a group of uniform materials based on organic salts (GUMBOS) have been introduced into the scientific literature involving many analytical, biological, and technological applications. In this regard, these nanoGUMBOS have been shown to display a number of unique properties including fluorescence, magnetism, tumor targeting, and optoelectronic. To date, however, little focus has been placed on developing and refining approaches for generation of size-controlled nanoGUMBOS from GUMBOS building blocks. Herein, we describe a systematic effort to define various strategies for the production of well-defined nanoGUMBOS. Specifically, we describe methods based on (i) sonochemical, (ii) microwave-assisted, (iii) cyclodextrin-assisted, and (iv) surfactant-assisted syntheses of nanoGUMBOS, evaluating the efficiency of each technique in controlling the size, sphericity, and uniformity of nanoGUMBOS produced. The effect of systematic variation in experimental parameters such as concentration, cation-to-anion ratio, as well as presence and type of template introduced for formation of nanoGUMBOS is also investigated.
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Ciclodextrinas/química , Nanopartículas/química , Compostos Orgânicos/química , Sais/química , Tensoativos/química , Fluorescência , Microscopia de Fluorescência , Micro-Ondas , Propriedades de SuperfícieRESUMO
A new kind of material called nanoGUMBOS, comprised entirely of cations and anions, has been developed by pairing various functional ions that exhibit fluorescence activity with biocompatible ions, in a process very much akin to that employed in ionic liquid chemistry. In the present study, spectral and biological properties of NIR absorbing nanoGUMBOS were evaluated using electron microscopy, dynamic light scattering, absorbance, thermal imaging, and live/dead fluorescence assays in conjunction with malignant MDA-MB-231 and non-malignant HS-578-BST epithelial human breast cells. The primary focus of this study was to maximize heat generation using NIR laser irradiation and minimize non-specific cytotoxicity using biocompatible constituent ions (e.g. amino acids, vitamins, or organic acids). Concurrently, in order to generate highly responsive nanomaterials for NIR-laser-triggered hyperthermia, optimization of the nanoparticle size, shape, and uniformity was carried out. Evaluation of data from hyperthermal studies of NIR absorbing nanoGUMBOS shows that these materials can achieve temperatures above the threshold for killing cancerous cells. Additionally, in vitro cell based assays demonstrated their promising hyperthermal effects on cancer derived epithelial cells.
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Nanoestruturas/química , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Feminino , Ácido Fólico/química , Humanos , Hipertermia Induzida , Lasers , Microscopia de Fluorescência , Nanoestruturas/toxicidadeRESUMO
Ionic liquid (IL) chemistry has evolved over the past century, such that these organic salts have impacted virtually every area of science and engineering. In the area of chemistry, initial applications of these salts were primarily the domain of chemists or chemical engineers who desired to manipulate the properties of IL solvents for a variety of applications including tuning various chemical processes. Since then, the chemistry of these organic salts has progressed such that changing an important property of a solvent (e.g., melting point or hydrophobicity) often involves simply altering the counterion of the organic salt. It is with this simplicity in mind that we have recently embarked upon the use of such chemistry to manipulate important properties of solid-phase ionic organic materials. To differentiate this chemistry from ionic liquid chemistry, we have coined the acronym GUMBOS (group of uniform materials based on organic salts). In this perspective article, we describe and demonstrate how ionic liquid chemistry can provide distinct and sometimes unique chemistry for solid-phase applications. Solid phase properties which can be manipulated via this chemistry include, but are not limited to, magnetism, melting point, hydrophobicity, fluorescence quantum yields, nanoformulations, material aggregation, viscosity, viscoelasticity, and cytotoxicity. In addition, we discuss a few examples to demonstrate how GUMBOS chemistry, until now, has been beneficial to the general area of materials chemistry and, more broadly, to the field of analytical chemistry. We also project future applications of this technology.
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The photothermal properties of several near-infrared-absorbing nanoparticles derived from group of uniform materials based on organic salts (GUMBOS) and composed of cationic dyes coupled with biocompatible anions are evaluated. These nanoparticles were synthesized using a reprecipitation method performed at various pH values: 2.0, 5.0, 7.0, 9.0, and 11.0. The cations for the nanoparticles derived from GUMBOS (nanoGUMBOS), [1048] and [1061], have absorbance maxima at wavelengths overlapping with human soft tissue absorbance minima. Near-infrared-absorbing nanoGUMBOS excited with a 1064 nm continuous laser led to heat generation, with an average temperature increase of 20.4 ± 2.7 °C. Although the [1061][Deoxycholate] nanoGUMBOS generated the highest temperature increase (23.7 ± 2.4 °C), it was the least photothermally efficient compound (13.0%) due to its relatively large energy band gap of 0.892 eV. The more photothermally efficient compound [1048][Ascorbate] (64.4%) had a smaller energy band gap of 0.861 eV and provided an average photothermal temperature increase of 21.0 ± 2.1 °C.
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OBJECTIVES: Reduction in faecal shedding of Shiga toxin-producing enterohaemorrhagic Escherichia coli (EHEC) in food-producing animals is a viable strategy to minimize human disease initiated by exposure to these microorganisms. To this end, an intervention strategy involving the electrostatic hybridization of two commonly used anti-infective agents for veterinary practice (i.e. chlorhexidine and ampicillin) was evaluated to curtail EHEC-transmitted disease from ruminant sources. Chlorhexidine di-ampicillin is a novel group of uniform material based on organic salts (GUMBOS) with inherent in vitro antibacterial activity that comes from its parent antimicrobial ions, chlorhexidine and ampicillin. METHODS: Antibacterial activities for chlorhexidine diacetate, sodium ampicillin, chlorhexidine di-ampicillin and stoichiometrically equivalent 1â:â2 chlorhexidine diacetateâ:âsodium ampicillin were assessed using the serial 2-fold dilution method and time-kill studies against seven isolates of E. coli O157:H7 and one non-pathogenic E. coli 25922. Further studies to investigate synergistic interactions of reacted and stoichiometrically equivalent unreacted antimicrobial agents at MICs and possible mechanisms were also investigated. RESULTS: Synergism and in vitro antibacterial activities against EHEC were observed in this study, which suggests chlorhexidine di-ampicillin could be a useful reagent in reducing EHEC transmission and minimizing EHEC-associated infections. Likewise, chlorhexidine di-ampicillin reduced HeLa cell toxicity as compared with chlorhexidine diacetate or the stoichiometric combination of antimicrobial agents. Further results suggest that the mechanisms of action of chlorhexidine di-ampicillin and chlorhexidine diacetate against E. coli O157:H7 are similar. CONCLUSIONS: Reacting antimicrobial GUMBOS as indicated in this study may enhance the approach to current combination drug therapeutic strategies for EHEC disease control and prevention.
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Ampicilina/uso terapêutico , Antibacterianos/uso terapêutico , Clorexidina/uso terapêutico , Desinfetantes/uso terapêutico , Infecções por Escherichia coli/prevenção & controle , Escherichia coli O157 , Antibacterianos/síntese química , Antibacterianos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Combinação de Medicamentos , Sinergismo Farmacológico , Quimioterapia Combinada , Microbiologia de Alimentos , Células HeLa , Humanos , Técnicas de Diluição do Indicador , Cinética , Testes de Sensibilidade Microbiana , Sais , Toxina Shiga/metabolismo , Escherichia coli Shiga Toxigênica/metabolismoRESUMO
A novel, simple and totally recyclable method has been developed for the synthesis of nontoxic, biocompatible and biodegradable composite materials from cellulose and chitosan. In this method, [BMIm+Cl-], an ionic liquid (IL), was used as a solvent to dissolve and synthesize the [CEL+CS] composite materials. Since the IL can be removed from the materials by washing them with water, and recovered from the washed solution, the method is totally recyclable. XRD, FTIR, NIR and SEM were used to characterize the materials and to confirm that CEL and CS were successfully regenerated by the method without any chemical transformation. More importantly, we have successfully demonstrated that [CEL+CS] material can serve as an effective adsorbent for removal of various endocrine disruptors including polychlorophenols and bisphenol A. This is because the composites have combined advantages of their components, namely superior chemical stability and mechanical stability (from CEL) and excellent adsorption capability for pollutants (from CS).
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We report on the synthesis and characterization of a PEGylated IR786 GUMBOS (Group of Uniform Materials Based on Organic Salts). The synthesis of this material was accomplished using a three step protocol: (1) substitution of chloride on the cyclohexenyl ring in the heptamethine chain of IR786 by 6-aminohexanoic acid, (2) grafting of methoxy polyethylene glycol (MeOPEG) onto the 6-aminohexanoic acid via an esterification reaction, and (3) anion exchange between [PEG786][I] and lithium bis(trifluoromethylsulfonyl)imide (LiNTf(2)) or sodium bis(2-ethylhexyl)sulfosuccinate (AOT) in order to obtain PEG786 GUMBOS. Examination of spectroscopic data for this PEG786 GUMBOS indicates a large stokes shift (122 nm). It was observed that this PEG786 GUMBOS associates in aqueous solution to form nano- and mesoscale self-assemblies with sizes ranging from 100 to 220 nm. These nano- and mesoscale GUMBOS are also able to resist nonspecific binding to proteins. PEGylation of the original IR786 leads to reduced cytotoxicity. In addition, it was noted that anions, such as NTf(2) and AOT, play a significant role in improving the photostability of PEG786 GUMBOS. Irradiation-induced J-aggregation in [PEG786][NTf(2)] and to some extent in [PEG786][AOT] produced enhanced photostability. This observation was supported by use of both steady state and time-resolved fluorescence measurements.
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Carbocianinas/química , Corantes/química , Ácido Dioctil Sulfossuccínico/química , Imidas/química , Indóis/química , Raios Infravermelhos , Nanopartículas/química , Polietilenoglicóis/química , Sulfonamidas/química , Adsorção , Ácido Aminocaproico/química , Animais , Bovinos , Ácido Dioctil Sulfossuccínico/toxicidade , Estabilidade de Medicamentos , Células HeLa , Humanos , Imidas/toxicidade , Nanopartículas/toxicidade , Soroalbumina Bovina/química , Solubilidade , Espectrometria de Fluorescência , Sulfonamidas/toxicidade , Água/químicaRESUMO
The ability to control the morphologies and spectral properties of organic low-dimensional nanomaterials is of paramount importance. The research reported herein demonstrates a template-free approach to tailored morphological and optical properties for a novel class of pseudoisocyanine (PIC)-based fluorescent organic nanoparticles derived from a group of uniform materials based on organic salts (GUMBOS). The synthesized nanoscale PIC-based particles (termed nanoGUMBOS), [PIC][NTf(2)] and [PIC][BETI], exhibit interesting adaptability as a function of the associated anion. The diamond-shaped nanostructures of [PIC][NTf(2)] and [PIC][BETI] nanorods exhibit enhanced fluorescence quantum yields relative to the parent compound, [PIC][I]. As supported by fluorescence lifetime measurements, these enhanced spectral properties can be attributed to differences in molecular self-assembly ordering (e.g., H- vs. J-aggregation) and restricted molecular rotation leading to reduced twisted intramolecular charge transfer in the nanoGUMBOS. The electrochemical properties of the PIC-based GUMBOS suggest their potential use in dye-sensitized solar cells.
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Ânions/química , Carbocianinas/química , Nanoestruturas/química , Fármacos Fotossensibilizantes/química , Técnicas Eletroquímicas , Polarização de Fluorescência , Teoria Quântica , Energia SolarRESUMO
Groups of uniform materials based on organic salts (GUMBOS), derived from thiacarbocyanine (TC)-based dyes with increasing methyne chain lengths, were prepared through a single-step metathesis reaction between the iodide form of the TC dye and lithium bis(perfluoroethylsulfonyl)imide as the lipophilic anion source. Ionic self-assembly of these fluorescent hydrophobic GUMBOS resulted in aqueous dispersions of one-dimensional micro-and nano-scale molecular aggregates. Blended binary and ternary aggregates containing multiple TC GUMBOS were also prepared. These nanostructures exhibited a variety of aspect ratios, affording tunable Förster resonance energy transfer (FRET) and aggregation-dependent spectroscopic properties.
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Microstructures of sodium deoxycholate hydrogels were altered considerably in the presence of variable tris(hydroxymethyl)aminomethane (TRIS) concentrations. These observations were confirmed by use of X-ray diffraction, polarized optical microscopy, rheology, and differential scanning calorimetry measurements. Our studies reveal enhanced gel crystallinity and rigidity with increasing TRIS concentrations. The tunable hydrogel microstructures obtained under various conditions have been successfully utilized as templates to synthesize cyanine-based fluorescent nanoGUMBOS (nanoparticles from a group of uniform materials based on organic salts). A systematic variation in size (70-200 nm), with relatively low polydispersity and tunable spectral properties of [HMT][AOT] nanoGUMBOS, was achieved by use of these modified hydrogels. The gel microstructures are observed to direct the size as well as molecular self-assembly of the nanomaterials, thereby tuning their spectral properties. These modified hydrogels were also found to possess other interesting properties such as variable morphologies ranging from fibrous to spherulitic, variable degrees of crystallinity, rigidity, optical activity, and release profiles which can be exploited for a multitude of applications. Hence, this study demonstrates a novel method for modification of sodium deoxycholate hydrogels, their applications as templates for nanomaterials synthesis, as well as their potential applications in biotechnology and drug delivery.
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Ácido Desoxicólico/química , Hidrogéis , Nanopartículas , Fluorescência , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Difração de Raios XRESUMO
Multifunctional phosphonium-lanthanide compounds that simultaneously possess paramagnetism, luminescence, and tumor mitochondrial targeting properties were prepared by use of a facile method. These compounds were fully characterized by use of (1)H, (13)C, (31)P NMR, FT-IR, and elemental analyses. The thermal properties of these compounds including melting points and decomposition temperatures were investigated using DSC and TGA analyses. In addition, the paramagnetism, luminescence, and tumor targeting properties of these multifunctional compounds were confirmed by respective use of SQUID, fluorescence, and cell cytotoxicity studies. All compounds exhibited paramagnetism at room temperature, which could provide target delivery of these compounds to parts of the body containing tumor cells using a strong external magnetic field. In addition, these compounds display two major characteristic emissions originating from Dy(3+), which can be utilized for imaging tumor cells. The IC(50) values of these compounds measured against normal breast cell line (Hs578Bst) are significantly greater than those measured against the corresponding carcinoma breast cell line (Hs578T), clearly indicating the selective tumor targeting properties of these compounds. Confocal fluorescence microscopy studies were used to confirm the yellowish-green fluorescence corresponding to the emission of dysprosium thiocyanate anion within cancer cells upon exposure of cancer cell lines such as human pancreatic carcinoma cell line (MIAPaCa-2) and human breast carcinoma (MDA-MB-231) to a solution of these phosphonium-dysprosium compounds.
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Antineoplásicos/síntese química , Sobrevivência Celular/efeitos dos fármacos , Complexos de Coordenação/síntese química , Disprósio/química , Oniocompostos/química , Compostos Organofosforados/química , Antineoplásicos/química , Antineoplásicos/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Complexos de Coordenação/química , Complexos de Coordenação/metabolismo , Complexos de Coordenação/farmacologia , Humanos , Concentração Inibidora 50 , Magnetometria , Microscopia Confocal , Microscopia de FluorescênciaRESUMO
The interactions of the negatively charged achiral molecular micelle, poly (sodium N-undecanoyl sulfate) (poly-SUS), with four different proteins using intrinsic and extrinsic fluorescence spectroscopic probes, are studied. A comparison of poly-SUS with the conventional surfactant, sodium dodecyl sulfate (SDS), and the monomeric species, SUS, is also reported. In this work, we observed that poly-SUS preferentially binds to acidic proteins, exhibiting positive cooperativity at concentrations less than 1 mM for all proteins studied. Moreover, it appears that the hydrophobic microdomain formed through polymerization of the terminal vinyl group of the monomer, SUS, is largely responsible for the superior binding capacity of poly-SUS. From these results, we conclude that the interactions of poly-SUS with the acidic proteins are predominantly hydrophobic and postulate that poly-SUS would produce superior interactions relative to SDS at low concentrations in polyacrylamide gel electrophoresis (PAGE). As predicted, use of poly-SUS allowed separation of the His-tagged tumor suppressor protein, p53, at sample buffer concentrations as low as 0.08% w/v (2.9 mM), which is 24 times lower than required for SDS in the standard reducing PAGE protocol. This work highlights the use of poly-SUS as an effective surfactant in 1D biochemical analysis.
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Quimotripsinogênio/química , Lactalbumina/química , Ovalbumina/química , Polivinil/química , Soroalbumina Bovina/química , Dodecilsulfato de Sódio/química , Ésteres do Ácido Sulfúrico/química , Micelas , Estrutura Molecular , Tamanho da Partícula , Polivinil/síntese química , Ésteres do Ácido Sulfúrico/síntese química , Propriedades de SuperfícieRESUMO
Herein we report the synthesis of a fluorescent organic salt through anion exchange and the subsequent fabrication of 1D-nanostructures via a facile templating method.
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Corantes Fluorescentes/química , Nanoestruturas/química , Sais/química , Óxido de Alumínio/química , Resinas de Troca Aniônica/química , Rodaminas/química , Espectrofotometria Ultravioleta , Tetrafenilborato/químicaRESUMO
Excessive production of reactive oxygen species can lead to alteration of cellular functions responsible for many diseases including cardiovascular diseases, neurodegenerative diseases, cancer, and aging. Hydroxyl radical is a short-lived radical which is considered very aggressive due to its high reactivity toward biological molecules. In this study, a COumarin-NEutral Red (CONER) nanoprobe was developed for detection of hydroxyl radical based on the ratiometric fluorescence signal between 7-hydroxy coumarin 3-carboxylic acid and neutral red dyes. Biocompatible poly lactide-co-glycolide (PLGA) nanoparticles containing encapsulated neutral red were produced using a coumarin 3-carboxylic acid conjugated poly(sodium N-undecylenyl-Nε-lysinate) (C3C-poly-Nε-SUK) as moiety reactive to hydroxyl radicals. The response of the CONER nanoprobe was dependent on various parameters such as reaction time and nanoparticle concentration. The probe was selective for hydroxyl radicals as compared with other reactive oxygen species including O(2)(â¢-), H(2)O(2), (1)O(2), and OCl(-). Furthermore, the CONER nanoprobe was used to detect hydroxyl radicals in vitro using viable breast cancer cells exposed to oxidative stress. The results suggest that this nanoprobe represents a promising approach for detection of hydroxyl radicals in biological systems.