Claudia Pacholski

Real-time monitoring of enzyme activity in a mesoporous silicon double layer

A double layer mesoporous silicon with different pore sizes functions as a nano-reactor that can isolate, filter and quantify the kinetics of enzyme reactions in real-time by optical reflectivity. This tiny reactor may be used to rapidly characterize a variety of isolated enzymes in a label-free manner. Activity of certain protease enzymes is often an indicator of disease states such as cancer1,2, stroke2, and neurodegeneracy3, and thus, there is a need for rapid assays that can characterize the kinetics and substrate specificity of enzymatic reactions. Nanostructured membranes can efficiently separate biomolecules4 but coupling a sensitive detection method remains difficult. Here we report a single mesoporous nano-reactor that can isolate and quantify in real-time the reaction products of proteases. The reactor consists of two layers of porous films electrochemically prepared from crystalline silicon. The upper layer with large pore sizes traps the protease enzymes and acts as the reactor while the lower layer with smaller pore sizes excludes the large proteins and captures the reaction products. Infiltration of the digested fragments into the lower layer produces a measurable change in optical reflectivity and this allows label-free quantification of enzyme kinetics in real-time within a volume of approximately 5 nanoliters.

Photonic Crystal Sensors Based on Porous Silicon

Porous silicon has been established as an excellent sensing platform for the optical detection of hazardous chemicals and biomolecular interactions such as DNA hybridization, antigen/antibody binding, and enzymatic reactions. Its porous nature provides a high surface area within a small volume, which can be easily controlled by changing the pore sizes. As the porosity and consequently the refractive index of an etched porous silicon layer depends on the electrochemial etching conditions photonic crystals composed of multilayered porous silicon films with well-resolved and narrow optical reflectivity features can easily be obtained. The prominent optical response of the photonic crystal decreases the detection limit and therefore increases the sensitivity of porous silicon sensors in comparison to sensors utilizing Fabry-Pérot based optical transduction. Development of porous silicon photonic crystal sensors which allow for the detection of analytes by the naked eye using a simple color change or the fabrication of stacked porous silicon photonic crystals showing two distinct optical features which can be utilized for the discrimination of analytes emphasize its high application potential.

Nanopatterning by block copolymer micelle nanolithography and bioinspired applications

This comprehensive overview of block copolymer micelle nanolithography (BCMN) will discuss the synthesis of inorganic nanoparticle arrays by means of micellar diblock copolymer approach and the resulting experimental control of individual structural parameters of the nanopattern, e.g., particle density and particle size. Furthermore, the authors will present a combinational approach of BCMN with conventional fabrication methods, namely, photolithography and electron beam lithography, which combines the advantages of high-resolution micronanopatterning with fast sample processing rates. In addition, the authors will demonstrate how these nanoparticle assemblies can be transferred to polymer substrates with a wide range of elasticity. In the second part of this report the authors will introduce some of the most intriguing applications of BCMN in biology and materials science: The authors will demonstrate how nanoparticle arrays may be used as anchor points to pattern functional proteins with single molecule resolution for studying cellular adhesion and present a technological roadmap to high-performance nanomaterials by highlighting recent applications for biomimetic optics and nanowires.

Self-assembled plasmonic core shell clusters with an isotropic magnetic dipole response in the visible range

We theoretically analyze, fabricate, and characterize a three-dimensional plasmonic nanostructure that exhibits a strong and isotropic magnetic response in the visible spectral domain. Using two different bottom-up approaches that rely on self-organization and colloidal nanochemistry, we fabricate clusters consisting of dielectric core spheres, which are smaller than the wavelength of the incident radiation and are decorated by a large number of metallic nanospheres. Hence, despite having a complicated inner geometry, such a core-shell particle is sufficiently small to be perceived as an individual object in the far field. The optical properties of such complex plasmonic core-shell particles are discussed for two different core diameters.

pH-triggered release of vancomycin from protein-capped porous silicon films

OBJECTIVE An in vitro model system for pH-triggered release of the antibiotic vancomycin from porous Si films is studied. METHOD Vancomycin is infused into a mesoporous Si film from a mixed aqueous/acetonitrile solution and trapped by a capping layer containing the protein bovine serum albumin (BSA). The protein effectively traps vancomycin in the porous nanostructure at pH 4.0; the protein dissolves and vancomycin is released into solution when the pH increases to 7.4. The surface chemistry of porous Si exerts a substantial effect on the efficacy of drug loading. The amount of drug loading is larger in freshly-etched (hydrophobic, hydrogen-terminated) porous Si and smaller in methyl-modified, undecylenic acid-modified and thermally oxidized samples. The quantity of drug loaded in a freshly etched porous Si chip is proportional to the thickness of the porous layer, which exhibits a constant volume loading efficiency of 31% (v/v). Flow-cell experiments designed to mimic the transition from pH 4 to 7 that occurs when material moves from the stomach to the upper intestinal tract were performed on the freshly etched films and vancomycin- and BSA-release rates were quantified from the effluent of the flow cell by high-pressure liquid chromatography analysis. RESULTS & CONCLUSION There is a small, constant rate of vancomycin release at pH 4 that is independent of the amount of drug loaded in the pores. This is attributed to diffusion of vancomycin from the BSA-capping layer. The release rate increases five- to tenfold when the pH of the solution in the flow cell increases to 7.4; 100% of the drug is released within 3 h of this increase.

Delivery of nanogram payloads using magnetic porous silicon microcarriers

A transport and delivery system for nanogram quantities of molecular species that does not use microfluidic channels, pumps, or valves is described. Microparticles consisting of magnetic porous silicon are prepared, and loading and delivery of an enzymatic payload are demonstrated. The high porosity (60%) porous Si host particles are made magnetic by infusion of superparamagnetic iron oxide nanoparticles (30 nm-diameter magnetite, Fe(3)O(4)) under oxidative conditions. After magnetite incorporation, the porous microparticle is still empty enough to accommodate nanogram quantities of a molecular payload; the enzymes horseradish peroxidase or pronase E are used in the present work. The assembly can be transported to a microliter water droplet containing the enzyme substrate with the aid of an external magnetic field. The enzyme is released into the droplet upon contact. The particles can be transported through air or a hydrocarbon liquid without loss in enzymatic activity of the payload.

Fabrication of porous silicon by metal-assisted etching using highly ordered gold nanoparticle arrays

A simple method for the fabrication of porous silicon (Si) by metal-assisted etching was developed using gold nanoparticles as catalytic sites. The etching masks were prepared by spin-coating of colloidal gold nanoparticles onto Si. An appropriate functionalization of the gold nanoparticle surface prior to the deposition step enabled the formation of quasi-hexagonally ordered arrays by self-assembly which were translated into an array of pores by subsequent etching in HF solution containing H2O2. The quality of the pattern transfer depended on the chosen preparation conditions for the gold nanoparticle etching mask. The influence of the Si surface properties was investigated by using either hydrophilic or hydrophobic Si substrates resulting from piranha solution or HF treatment, respectively. The polymer-coated gold nanoparticles had to be thermally treated in order to provide a direct contact at the metal/Si interface which is required for the following metal-assisted etching. Plasma treatment as well as flame annealing was successfully applied. The best results were obtained for Si substrates which were flame annealed in order to remove the polymer matrix - independent of the substrate surface properties prior to spin-coating (hydrophilic or hydrophobic). The presented method opens up new resources for the fabrication of porous silicon by metal-assisted etching. Here, a vast variety of metal nanoparticles accessible by well-established wet-chemical synthesis can be employed for the fabrication of the etching masks.

Contact Line Motion on Nanorough Surfaces: A Thermally Activated Process

The motion of a solid-liquid-liquid contact line over nanorough surfaces is investigated. The surface nanodefects are varied in size, density, and shape. The dynamics of the three-phase contact line on all nanorough substrates studied is thermally activated. However, unlike the motion of a liquid-vapor interface over smooth surfaces, this thermally activated process is not adequately described by the molecular kinetic theory. The molecular parameters extracted from the experiments suggest that on the nanorough surfaces, the motion of the contact line is unlikely to simply consist of molecular adsorption-desorption steps. Thermally activated pinning-depinning events on the surface nanodefects are also important. We investigate the effect of surface nanotopography on the relative importance of these two mechanisms in governing contact line motion. Using a derivation for the hysteresis energy based on Joanny and de Genness model, we evaluate the effect of nanotopographical features on the wetting activation free energy and contact line friction. Our results suggest that both solid-liquid interactions and surface pinning strength contribute to the energy barriers hindering the three-phase contact line motion. For relatively low nanodefect densities, the activation free energy of wetting can be expressed as a sum of surface wettability and surface topography contributions, thus providing a direct link between contact line dynamics and roughness parameters.

Simulating different manufactured antireflective sub-wavelength structures considering the influence of local topographic variations

Laterally structured antireflective sub-wavelength structures show unique properties with respect to broadband performance, damage threshold and thermal stability. Thus they are superior to classical layer based antireflective coatings for a number of applications. Dependent on the selected fabrication technology the local topography of the periodic structure may deviate from the perfect repetition of a sub-wavelength unit cell. We used rigorous coupled-wave analysis (RCWA) to simulate the efficiency losses due to scattering effects based on height and displacement variations between the individual protuberances. In these simulations we chose conical and Super-Gaussian shapes to approximate the real profile of fabricated structures. The simulation results are in accordance with the experimentally determined optical properties of sub-wavelength structures over a broad wavelength range. Especially the transmittance reduction in the deep-UV could be ascribed to these variations in the sub-wavelength structures.

Enhanced sputter yields of ion irradiated Au nano particles: energy and size dependence

Hexagonally arranged Au nanoparticles exhibiting a broad Gaussian-shaped size distribution ranging from 30 nm to 80 nm were deposited on Si substrates and irradiated with Ar(+) and Ga(+) ions with various energies from 20 to 350 keV and 1 to 30 keV, respectively. The size and energy dependence of the sputter yield were measured using high-resolution scanning electron microscopy image analysis. These results were compared to simulation results obtained by iradina, a Monte Carlo code, which takes the specifics of the nano geometry into account. The experimental sputter yields are significantly higher than simulated sputter yields for both bulk and the nano geometry. The difference can be clearly attributed to thermally driven effects, which significantly increase the measured sputter yields.