Hybrid nanogels by direct mixing of chitosan, tannic acid and magnetite nanoparticles: processes involved in their formation and potential catalytic properties.

Magnetite (Fe3O4) nanoparticles (MNPs) as nanocatalysts have drawn considerable attention because of their unique properties such as peroxidase-like activity. However, their biodistribution and availability for specific treatments still need to be improved. In this study, a simple and convenient strategy for the synthesis of hybrid nanogels (NGs) is described, which involves direct mixing of biomaterials such as chitosan (Ch) and tannic acid (TA), with the incorporation of MNPs, under oxidising conditions, using the inverse nanoemulsion method. The different processes involved in the formation of these hybrid nanosystems as well as their morphological and chemical structure are investigated using optical, spectroscopic, and electron microscopic techniques (DLS, UV-VIS, FT-IR, XPS, TEM, and SEM-EDS). It is demonstrated that ∼11 nm synthesized MNPs, post-functionalized with oxidised TA, act as covalent crosslinkers. As a proof of concept, the potential use of these materials in nanocatalytic medicine was evaluated using a colorimetric method based on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) in hydrogen peroxide. The results show that these hybrid nanogels have the same peroxidase-like activity as bare MNPs, indicating that the organic nanostructure stabilises the inorganic nanoparticles without any significant change in the catalytic properties. Therefore, this kind of nanomaterial has promising potential for use in nanocatalytic medicine with improved biocompatibility and biodistribution.

Colloidal gold clusters formation and chemometrics for direct SERS determination of bioanalytes in complex media.

In this work, we report the sensitive and selective sensing of the purine bases adenine and guanine in urine matrix by using surface-enhanced Raman spectroscopy (SERS) and a colloidal SERS substrate. To identify suitable conditions for quantitative analysis, the pH dependence of spectra of adenine, guanine, urine simulant and their mixtures was studied on gold nanoparticles suspension. Interestingly, although the urine matrix promotes the analytes signal suppression and overlapping bands, it can also cause an improvement in repeatability of the SERS measurements. This effect was associated to the relatively controlled formation of small-sized gold clusters and it was investigated both experimentally and theoretically. Furthermore, a correlation constrained multivariate curve resolution-alternating least squares (MCR-ALS) method was developed to resolve overlapping SERS bands and to quantify physiologically relevant (micromolar) concentrations of the bioanalytes. The performance of the proposed MCR-ALS approach (assessed in terms of figures of merit) was similar to that obtained by using partial least squares regression, but with the additional advantage of retrieving valuable spectral information. Therefore, this method can be used for improving selectivity of colloidal clusters in qualitative and quantitative SERS analysis of complex media, avoiding the need for tedious nanoparticle-surface modification or preliminary chromatographic separation.

A Plasmonic Approach to Study Protein Interaction Kinetics through the Dimerization of Functionalized Ag Nanoparticles.

Understanding the kinetics of protein interactions plays a key role in biology with significant implications for the design of analytical methods for disease monitoring and diagnosis in medical care, research and industrial applications. Herein, we introduce a novel plasmonic approach to study the binding kinetics of protein-ligand interactions following the formation of silver nanoparticles (Ag NPs) dimers by UV-Vis spectroscopy that can be used as probes for antigen detection and quantification. To illustrate and test the method, the kinetics of the prototype biotin-streptavidin (Biot-STV) pair interaction was studied. Controlled aggregates (dimers) of STV functionalized Ag NPs were produced by adding stoichiometric quantities of gliadin-specific biotinylated antibodies (IgG-Biot). The dimerization kinetics was studied in a systematic way as a function of Ag NPs size and at different concentrations of IgG-Biot. The kinetics data have shown to be consistent with a complex reaction mechanism in which only the Ag NPs attached to the IgG-Biot located in a specific STV site are able to form dimers. These results help in elucidating a complex reaction mechanism involved in the dimerization kinetics of functionalized Ag NPs, which can serve as probes in surface plasmon resonance-based bioassays for the detection and quantification of different biomarkers or analytes of interest.

Nanoparticle-cell interactions induced apoptosis: a case study with nanoconjugated epidermal growth factor.

In addition to the intrinsic toxicity associated with the chemical composition of nanoparticles (NP) and their ligands, biofunctionalized NP can perturb specific cellular processes through NP-cell interactions and induce programmed cell death (apoptosis). In the case of the epidermal growth factor (EGF), nanoconjugation has been shown to enhance the apoptotic efficacy of the ligand, but the critical aspects of the underlying mechanism and its dependence on the NP morphology remain unclear. In this manuscript we characterize the apoptotic efficacy of nanoconjugated EGF as a function of NP size (with sphere diameters in the range 20-80 nm), aspect ratio (A.R., in the range of 4.5 to 8.6), and EGF surface loading in EGFR overexpressing MDA-MB-468 cells. We demonstrate a significant size and morphology dependence in this relatively narrow parameter space with spherical NP with a diameter of approx. 80 nm being much more efficient in inducing apoptosis than smaller spherical NP or rod-shaped NP with comparable EGF loading. The nanoconjugated EGF is found to trigger an EGFR-dependent increase in cytoplasmic reactive oxygen species (ROS) levels but no indications of increased mitochondrial ROS levels or mitochondrial membrane damage are detected at early time points of the apoptosis induction. The increase in cytoplasmic ROS is accompanied by a perturbation of the intracellular glutathione homeostasis, which represents an important check-point for NP-EGF mediated apoptosis. Abrogation of the oxidative stress through the inhibition of EGFR signaling by the EGFR inhibitor AG1478 or addition of antioxidants N-acetyl cysteine (NAC) or tempol, but not trolox, successfully suppressed the apoptotic effect of nanoconjugated EGF. A model to account for the observed morphology dependence of EGF nanoconjugation enhanced apoptosis and the underlying NP-cell interactions is discussed.

Enzyme-Free Immunoassay Using Silver Nanoparticles for Detection of Gliadin at Ultralow Concentrations.

Determination of biomarkers in clinical or food samples is of crucial importance for monitoring, prevention, and care of public health. The standard procedure used for this purpose is the enzyme-linked immunosorbent assay (ELISA), which makes use of the specific antibody-antigen biorecognition and the catalytic effect of the enzymes. One of the main shortcomings of this technique is the use of enzymes that often present low chemical and thermal stabilities compared to other chemicals. Other drawbacks include the nonspecific binding process that could lead to false-positive results, the use of relatively large amounts of the sample, and the number of time-consuming steps involved. Recently, an enzyme-free and ultrasensitive analytical method for antigen detection denoted as intensity depletion immunolinked assay (IDILA) has been proposed by our laboratory. The assay is based on the inhibition to form Ag nanosphere dimers linked by a specific antibody in the presence of the corresponding antigen. In this work, we go a step further demonstrating how the performance of this method could be improved by using silver nanoparticles (Ag NPs) of different diameters (58 and 78 nm). The experiments are performed for detecting gliadin, an antigen of utmost importance in celiac disease, and the results are compared with ELISA, the standard technique homologated by the Food Codex Alimentarius. It is found that the IDILA assay could be around 1000 or 10 000 times more sensitive than ELISA, also having lower limits of detection, depending on the conditions explored (fraction of dimers and Ag NP diameter). Using the appropriate conditions, the IDILA assay is shown to be able to detect femtomolar concentrations of the antigen, besides being robust, reliable, cheap, rapid (around 2 h), and of easy implementation using the standard equipment and biomolecular reagents used for the ELISA assay.

Design of a novel plasmonic nanoconjugated analytical tool for ultrasensitive antigen quantification.

To date, while various diagnostic approaches for antigen detection have been proposed, most are too expensive, lengthy and limited in sensitivity for clinical use. Nanoparticle systems with unique material properties, however, circumvent these problems and offer improved accuracy and sensitivity over current methods like the enzyme-linked immunosorbent assay (ELISA). Herein, we present a novel functionalization strategy of plasmonic nanoparticle probes capable of specific quantification of antigens directly in clinical samples. A nanoconjugation strategy that allows one to perform an intensity depletion immuno-linked assay (IDILA), involving specific antibodies that target the antigen of interest was designed to obtain a calibration curve and achieve the quantification of the antigen in clinical samples in the same experiment using a microplate reader (i.e., an UV-vis spectrophotometer). Finally, the IDILA methodology allowed specific detection of various clinically relevant antigens, with significantly improved sensitivity over the ELISA. Furthermore, the assay was shown to be robust, reliable, cheap and rapid, diagnosing antigens in clinical serum samples within 2 hours.

Plasmonic interactions: from molecular plasmonics and Fano resonances to ferroplasmons.

Plasmon interactions are a subject of great interest from both the technological as well as the fundamental points of view. In this Perspective, we outline the great variety of physical phenomena that are produced by the interactions of localized surface plasmon resonance with molecular excitons; with other plasmonic nanostructures, particularly the Fano effect; and with nonplasmonic nanoparticles, such as the just-reported interaction with ferromagnetic nanoparticles. The theoretical as well as experimental challenges remaining to be elucidated are discussed.

Identification, localization, and quantification of neuronal cell membrane receptors with plasmonic probes: role of protein kinase D1 in their distribution.

Detecting, imaging, and being able to localize the distribution of several cell membrane receptors on a single neuron are very important topics in neuroscience research. In the present work, the distribution of metabotropic glutamate receptor 1a (mGluR1a) density on neuron cells on subcellular length scales is determined by evaluating the role played by protein kinase D1 (PKD1) in the trafficking of membrane proteins, comparing the distribution of mGluR1a in experiments performed in endogenous PKD1 expression with those in the presence of kinase-inactive protein kinase D1 (PKD1-kd). The localization, distribution, and density of cell surface mGluR1a were evaluated using 90 nm diameter Au nanoparticle (NP) probes specifically functionalized with a high-affinity and multivalent labeling function, which allows not only imaging NPs where this receptor is present but also quantifying by optical means the NP density. This is so because the NP generates a density (ρ)-dependent SERS response that facilitated a spatial mapping of the mGluR1a density distribution on subcellular length scales (dendrites and axons) in an optical microscope. The measured ρ values were found to be significantly higher on dendrites than on axons for endogenous PKD1, while an increase of ρ on axons was observed when PKD1 is altered. The spatial distribution of the NP immunolabels through scanning electron microscopy (SEM) confirmed the results obtained by fluorescence bright-field analysis and dark-field spectroscopy and provided additional structural details. In addition, it is shown using electrodynamic simulations that SERS spectroscopy could be a very sensitive tool for the spatial mapping of cell membrane receptors on subcellular length scales, as SERS signals are almost linearly dependent on NP density and therefore give indirect information on the distribution of cell membrane proteins. This result is important since the calibration of the ρ-dependent near-field enhancement of the Au immunolabels through correlation of SERS and SEM paves the way toward quantitative immunolabeling studies of cell membrane proteins involved in neuron polarity. From the molecular biology point of view, this study shows that in cultured hippocampal pyramidal cells mGluR1a is predominantly transported to dendrites and excluded from axons. Expression of kinase-inactive protein kinase D1 (PKD1-kd) dramatically and selectively alters the intracellular trafficking and membrane delivery of mGluR1a-containing vesicles.

One-step/one-pot decoration of oxide microparticles with silver nanoparticles.

HYPOTHESIS: Heterogeneous nucleation of silver oxide (Ag2O) onto oxide microparticles (OMPs) followed by spontaneous thermal decomposition produce nanostructures made of OMPs decorated with silver nanoparticles (OMP|AgNPs). EXPERIMENTS: Colloidal chemistry methods have been used to produce the decoration of OMPs with silver nanoparticles (AgNPs), by carrying out the Ag2O precipitation/thermal decomposition. The process is driven in water enriched acetone medium containing NaOH, NH3, AgNO3 and SiO2MPs as substrate. Optical and morphological properties of OMP|AgNPs were characterized by using STEM, EDS, HRTEM and Raman spectroscopy. FINDINGS: A new synthetic method to decorate OMPs (TiO2, SiO2) with metallic AgNPs in a single step/single pot reaction is proven effective to produce OMP|AgNPs either in aqueous or water enriched media.

Quantitative understanding of the optical properties of a single, complex-shaped gold nanoparticle from experiment and theory.

We report on a combined study of Rayleigh and Raman scattering spectroscopy, 3D electron tomography, and discrete dipole approximation (DDA) calculations of a single, complex-shaped gold nanoparticle (NP). Using the exact reconstructed 3D morphology of the NP as input for the DDA calculations, the experimental results can be reproduced with unprecedented precision and detail. We find that not only the exact NP morphology but also the surroundings including the points of contact with the substrate are of crucial importance for a correct prediction of the NP optical properties. The achieved accuracy of the calculations allows determining how many of the adsorbed molecules have a major contribution to the Raman signal, a fact that has important implications for analyzing experiments and designing sensing applications.

Quantum dynamical simulations of local field enhancement in metal nanoparticles.

Field enhancements (Γ) around small Ag nanoparticles (NPs) are calculated using a quantum dynamical simulation formalism and the results are compared with electrodynamic simulations using the discrete dipole approximation (DDA) in order to address the important issue of the intrinsic atomistic structure of NPs. Quite remarkably, in both quantum and classical approaches the highest values of Γ are located in the same regions around single NPs. However, by introducing a complete atomistic description of the metallic NPs in optical simulations, a different pattern of the Γ distribution is obtained. Knowing the correct pattern of the Γ distribution around NPs is crucial for understanding the spectroscopic features of molecules inside hot spots. The enhancement produced by surface plasmon coupling is studied by using both approaches in NP dimers for different inter-particle distances. The results show that the trend of the variation of Γ versus inter-particle distance is different for classical and quantum simulations. This difference is explained in terms of a charge transfer mechanism that cannot be obtained with classical electrodynamics. Finally, time dependent distribution of the enhancement factor is simulated by introducing a time dependent field perturbation into the Hamiltonian, allowing an assessment of the localized surface plasmon resonance quantum dynamics.

Preparation of controlled gold nanoparticle aggregates using a dendronization strategy.

In this work, a dendronization strategy was used to control interparticle spacing and the optical properties of gold nanoparticle (NP) aggregates in aqueous media. To achieve this goal, two dendritic disulfides bearing different functionalities on their periphery were synthesized and used as ligands to dendronize gold NPs. The dendronized NPs then undergo aggregation; this process was followed by UV-vis spectroscopy, dynamic light scattering (DLS), and transmission electronic microscopy (TEM) measurements and correlated with Generalized Mie Theory electrodynamics calculations. For comparison, NP functionalization was also studied using a nondendritic ligand. It was found that the use of dendritic disulfides allows for the preparation of controlled NP aggregates. This study demonstrates how different dendronization parameters, such as disulfide concentration, temperature, time and nature of the ligand (dendritic vs nondendritic), determine the control exerted over the size and stability of the NP aggregates.

Rational design of plasmonic nanostructures for biomolecular detection: interplay between theory and experiments.

In this work, we report a simple strategy to obtain ultrasensitive SERS nanostructures by self-assembly and bioconjugation of Au nanospheres (NSs). Homodimer aggregates with an interparticle gap of around 8 nm are generated in aqueous dispersions by the highly specific molecular recognition of biotinylated Au NSs to streptavidin (STV), while random Au NS aggregates with a gap of 5 nm are formed in the absence of STV due to hydrogen bonding among biotinylated NSs. Both types of aggregates depict SERS analytical enhancement factors (AEF) of around 10(7) and the capability to detect biotin concentrations lower than 1 × 10(-12) M. Quite interesting, the AEF for an external analyte, Rhodamine 6G (RH6G), using the dimer aggregates is 1 order of magnitude greater (10(5)) than using random aggregates (around 10(4)). The dependence on the wavelength and the differences of the AEF for Au random aggregates and dimers are rationalized with rigorous electrodynamic simulations. The dimers obtained afford a new type of an in situ self-calibrated and reliable SERS substrate where biotinylated molecules can selectively be "trapped" by STV and located in the nanogap enhanced plasmonic field. Using this concept, powerful molecular-recognition-based SERS assays can be carried out. The capability of the dimeric structures for analytical applications is demonstrated using SPR spectroscopy to detect biotinylated immunoglobulin G at very low concentrations.

Retrieving the spatial distribution of cavity modes in dielectric resonators by near-field imaging and electrodynamics simulations.

For good performance of photonic devices whose working principle is based on the enhancement of electromagnetic fields obtained by confining light into dielectric resonators with dimensions in the nanometre length scale, a detailed knowledge of the optical mode structure becomes essential. However, this information is usually lacking and can only be indirectly obtained by conventional spectroscopic techniques. Here we unraveled the influence of wire size, incident wavelength, degree of polarization and the presence of a substrate on the optical near fields generated by cavity modes of individual hexagonal ZnO nanowires by combining scanning near-field optical microscopy (SNOM) with electrodynamics calculations within the discrete dipole approximation (DDA). The near-field patterns obtained with very high spatial resolution, better than 50 nm, exhibit striking size and spatial-dispersion effects, which are well accounted for within DDA, using a wavevector-dependent dipolar interaction and considering the dielectric anisotropy of ZnO. Our results show that both SNOM and DDA simulations are powerful tools for the design of optoelectronic devices able to manipulate light at the nanoscale.

Optical properties of metallic nanoparticles: manipulating light, heat and forces at the nanoscale.

We present to a general readership an overview of the rich variety of phenomena and applications that arise from the interaction of metallic nanoparticles with light. First, we present the fundamental physics of localized surface plasmon resonances, the most relevant theories and numerical methods, as well as optical detection schemes. Finally, we explain how the localized surface plasmon resonances are currently exploited for the nanoscale manipulation of light, heat and forces in various applications and experimental investigations.

Using highly accurate 3D nanometrology to model the optical properties of highly irregular nanoparticles: a powerful tool for rational design of plasmonic devices.

The realization of materials at the nanometer scale creates new challenges for quantitative characterization and modeling as many physical and chemical properties at the nanoscale are highly size and shape-dependent. In particular, the accurate nanometrological characterization of noble metal nanoparticles (NPs) is crucial for understanding their optical response that is determined by the collective excitation of conduction electrons, known as localized surface plasmons. Its manipulation gives place to a variety of applications in ultrasensitive spectroscopies, photonics, improved photovoltaics, imaging, and cancer therapy. Here we show that by combining electron tomography with electrodynamic simulations an accurate optical model of a highly irregular gold NP synthesized by chemical methods could be achieved. This constitutes a novel and rigorous tool for understanding the plasmonic properties of real three-dimensional nano-objects.

Near-field enhancement of multipole plasmon resonances in Ag and Au nanowires.

In this paper, we investigate theoretically the electromagnetic field enhancement arising from excitation of silver and gold nanowires (NWs) of finite length, capable of sustaining surface plasmon resonances of different multipole order, using the Discrete Dipole Approximation (DDA). The influence of NW length on the degree of enhancement and confinement of the electromagnetic field for each surface plasmon mode is analyzed by a 3D mapping of the near field for different planes around the NW as well by calculating its variation with distance along two different directions, one parallel to and the other perpendicular to the NW axis, outside of the NW. It was found that the enhancement is still significant at relative large distances from the NW end, its decay being of much longer range than that predicted by a simple dipole approximation, especially at near-infrared wavelengths.

Accounting for the dependence of P(E',E) on the maximum impact parameter in classical trajectory calculations: application to the H2O-H2O collisional relaxation.

In this work we report a novel methodology that is able to predict how energy transfer transition probability density functions [P(E',E)] change with the maximum impact parameter (bmax) used in trajectory calculations (TC's). The method assumes that P(E',E) can be described by a sum of exponential functions and that all the trajectories with an initial impact parameter beyond a certain critical value will contribute only to the elastic peak [P(E',E) for E'=E]. This approach is applied to H2O-H2O collisions at different initial vibrational energies of the excited molecules and temperatures of bath gas. The results show that it is possible to reproduce with high accuracy the whole P(E',E) obtained from a given bmax, using the results of TC's performed at another bmax. The new methodology also leads us to propose a new criterion to choose the value of bmax.

Building transition probabilities for any condition using reduced cumulative energy transfer functions in H2O-H2O collisions.

The energy transfer process between highly vibrationally excited H(2)O in thermal equilibrium with a gas bath of H(2)O at different internal energies and temperatures has been studied by classical trajectory calculations. The results were analyzed using a cumulative probability distribution Q(DeltaE) of the amount of energy transferred, obtained by direct count of the number of trajectories that transfer an amount of energy equal to or greater than a certain value DeltaE. Scaling Q(DeltaE) in terms of the mean down and up energies transferred for each group of trajectories results in a unique distribution. This fact and the use of detailed balance constrains were used to propose a methodology that make it possible to build the whole P(E('),E) for any condition by knowing DeltaE and a series of parameters that depend only on the system under study.

Fitting complex potential energy surfaces to simple model potentials: application of the simplex-annealing method.

A stochastic method of optimization, which combines simulated annealing with simplex, is implemented to fit the parameters of a simple model potential. The main characteristic of the method is that it explores the whole space of the parameters of the model potential, and therefore it is very efficient in locating the global minimum of the cost function, in addition to being independent of the initial guess of the parameters. The method is employed to fit the complex intermolecular potential energy surface of the dimer of water, using as a reference the spectroscopic quality anisotropic site-site potential of Feller et al. The simple model potential chosen for its reparameterization is the MCY model potential of Clementi et al. The quality of the fit is assessed by comparing the geometry of the minimum, the harmonic frequencies, and the second virial coefficients of the parameterized potential with the reference one. Finally, to prove more rigorously the robustness of this method, it is compared with standard nonstochastic methods of optimization.