Optical Monitoring of the Capillary Filling Dynamics Variation in Nanoporous Anodic Alumina toward Sensing Applications.

Fluid imbibition-coupled laser interferometry (FICLI) is a technique in which the kinetics of a fluid infiltrating a nanoporous anodic alumina (NAA) membrane is monitored by the interference of a laser beam at the membrane top and bottom surfaces. Further processing of the measured data results in an estimate of the pore radius. In this work, we study the accuracy of FICLI in the detection of small changes in pore radius, and we evaluate the possibility of using such detection as a sensing paradigm. The accuracy is estimated by measuring samples with increasing pore radius, obtained by successive wet etching steps, and repeatability is evaluated by using different liquids. For decreasing pore radius, samples obtained by the successive deposition of polyelectrolyte double layers are used. With the aim of evaluating the possibility of the FICLI method to sense biological binding events, BSA attachment detection is demonstrated by applying FICLI to samples before and after immobilization of the protein. Results show that the technique permits an accurate estimation of the pore radius, the pore-etching rate (with a radius variation of retch,DI = 1.05 nm/min ± 0.11 nm/min), and the polyelectrolyte double layer thickness (with a radius variation of rPAH/PSS = 3.2 nm ± 0.2 nm per polyelectrolyte double layer). Furthermore, the pore radius reduction measured after BSA immobilization (dBSA = 4.9 nm ± 1.1 nm) is in good agreement with the protein size, as reported in the literature. With these results, we provide a sound basis for the applicability of FICLI as a sensitive technique for the characterization of NAA pore radius modifications.

Recent Advances in Nanoporous Anodic Alumina: Principles, Engineering, and Applications.

The development of aluminum anodization technology features many stages. With the story stretching for almost a century, rather straightforward-from current perspective-technology, raised into an iconic nanofabrication technique. The intrinsic properties of alumina porous structures constitute the vast utility in distinct fields. Nanoporous anodic alumina can be a starting point for: Templates, photonic structures, membranes, drug delivery platforms or nanoparticles, and more. Current state of the art would not be possible without decades of consecutive findings, during which, step by step, the technique was more understood. This review aims at providing an update regarding recent discoveries-improvements in the fabrication technology, a deeper understanding of the process, and a practical application of the material-providing a narrative supported with a proper background.

Supported Ultra-Thin Alumina Membranes with Graphene as Efficient Interference Enhanced Raman Scattering Platforms for Sensing.

The detection of Raman signals from diluted molecules or biomaterials in complex media is still a challenge. Besides the widely studied Raman enhancement by nanoparticle plasmons, interference mechanisms provide an interesting option. A novel approach for amplification platforms based on supported thin alumina membranes was designed and fabricated to optimize the interference processes. The dielectric layer is the extremely thin alumina membrane itself and, its metallic aluminum support, the reflecting medium. A CVD (chemical vapor deposition) single-layer graphene is transferred on the membrane to serve as substrate to deposit the analyte. Experimental results and simulations of the interference processes were employed to determine the relevant parameters of the structure to optimize the Raman enhancement factor (E.F.). Highly homogeneous E.F. over the platform surface are obtained, typically 370 ± (5%), for membranes with ~100 nm pore depth, ~18 nm pore diameter and the complete elimination of the Al2O3 bottom barrier layer. The combined surface enhanced Raman scattering (SERS) and interference amplification is also demonstrated by depositing ultra-small silver nanoparticles. This new approach to amplify the Raman signal of analytes is easily obtained, low-cost and robust with useful enhancement factors (~400) and allows only interference or combined enhancement mechanisms, depending on the analyte requirements.

Tunable Nanoporous Anodic Alumina Photonic Crystals by Gaussian Pulse Anodization.

This study presents a Gaussian pulse anodization approach to generate nanoporous photonic crystals with highly tunable and controllable optical properties across the visible-NIR spectrum. Nanoporous anodic alumina Gaussian photonic crystals (NAA-GPCs) are fabricated in oxalic acid electrolyte by Gaussian pulse anodization, a novel form of pulse-like anodization. The effect of the Gaussian pulse width in the anodization profile on the optical properties of these photonic crystals is assessed by systematically varying this fabrication parameter from 5 to 60 s. The optical features of the characteristic photonic stopband (PSB) of NAA-GPCs-the position of the central wavelength, full width at half-maximum, and intensity-are found to be highly dependent on the Gaussian pulse width, the angle of incidence of incoming photons, and the nanopore diameter of NAA-GPCs. The effective medium of NAA-GPCs is assessed by monitoring spectral shifts in their characteristic PSB upon infiltration of their nanoporous structure with analytical solutions of d-glucose of varying concentration (0.0125-1 M). Experimental results are validated and mechanistically described by theoretical simulations, using the Looyenga-Landau-Lifshitz effective medium approximation model. Our findings demonstrate that Gaussian pulse anodization is an effective nanofabrication approach to producing highly sensitive NAA-based PC structures with versatile and tunable PSBs across the spectral regions. The findings provide new exiting opportunities to integrate these unique PC structures into photonic sensors and other platform materials for light-based technologies.

Real-Time Monitoring of Biotinylated Molecules Detection Dynamics in Nanoporous Anodic Alumina for Bio-Sensing.

The chemical modification, or functionalization, of the surfaces of nanomaterials is a key step to achieve biosensors with the best sensitivity and selectivity. The surface modification of biosensors usually comprises several modification steps that have to be optimized. Real-time monitoring of all the reactions taking place during such modification steps can be a highly helpful tool for optimization. In this work, we propose nanoporous anodic alumina (NAA) functionalized with the streptavidin-biotin complex as a platform towards label-free biosensors. Using reflective interferometric spectroscopy (RIfS), the streptavidin-biotin complex formation, using biotinylated thrombin as a molecule model, was monitored in real-time. The study compared the performance of different NAA pore sizes in order to achieve the highest response. Furthermore, the optimal streptavidin concentration that enabled the efficient detection of the biotinylated thrombin attachment was estimated. Finally, the ability of the NAA-RIfS system to quantify the concentration of biotinylated thrombin was evaluated. This study provides an optimized characterization method to monitor the chemical reactions that take place during the biotinylated molecules attachment within the NAA pores.

Stacked Nanoporous Anodic Alumina Gradient-Index Filters with Tunable Multispectral Photonic Stopbands as Sensing Platforms.

This study presents the development and optical engineering of stacked nanoporous anodic alumina gradient-index (NAA-GIFs) filters with tunable multispectral photonic stopbands for sensing applications. The structure of these photonic crystals (PC) is formed by stacked layers of NAA produced with sinusoidally modified effective medium. The progressive modification of the sinusoidal period during the anodization process enables the generation and precise tuning of the characteristic photonic stopbands (PSB) (i.e., one per sinusoidal period in the anodization profile) of these PC structures. Four types of NAA-GIFs featuring three distinctive PSBs positioned within the visible spectral region are developed. The sensitivity of the effective medium of these NAA-GIFs is systematically assessed by measuring spectral shifts in the characteristic PSBs upon infiltration of their nanoporous structure with analytical solutions of d-glucose with several concentrations (0.025-1 M). This study provides new insights into the intrinsic relationship between the nanoporous architecture of these PCs and their optical properties, generating opportunities to fabricate advanced optical sensing systems for high-throughput and multiplexed detection of analytes in a single sensing platform.

Selective and Sensitive Probe Based in Oligonucleotide-Capped Nanoporous Alumina for the Rapid Screening of Infection Produced by Candida albicans.

A robust, sensitive, and time-competitive system to detect Candida albicans in less than 30 min in clinical samples based in capped nanoporous anodic alumina (NAA) is developed. In the proposed design, NAA pores are loaded with rhodamine B and then blocked with an oligonucleotide that is able to recognize C. albicans DNA. The capped material shows negligible cargo release, whereas dye delivery is selectively accomplished when genomic DNA from C. albicans is present. This procedure has been successfully applied to detect C. albicans in clinical samples from patients infected with this yeast. When compared with classical C. albicans detection methods, the proposed probe has a short assay time, high sensitivity and selectivity, demonstrating the high potential of this simple design for the diagnosis of infection produced by C. albicans.

Nanoporous Anodic Alumina Surface Modification by Electrostatic, Covalent, and Immune Complexation Binding Investigated by Capillary Filling.

The fluid imbibition-coupled laser interferometry (FICLI) technique has been applied to detect and quantify surface changes and pore dimension variations in nanoporous anodic alumina (NAA) structures. FICLI is a noninvasive optical technique that permits the determination of the NAA average pore radius with high accuracy. In this work, the technique is applied after each step of different surface modification paths of the NAA pores: (i) electrostatic immobilization of bovine serum albumin (BSA), (ii) covalent attachment of streptavidin via (3-aminipropyl)-triethoxysilane and glutaraldehyde grafting, and (iii) immune complexation. Results show that BSA attachment can be detected as a reduction in estimated radius from FICLI with high accuracy and reproducibility. In the case of the covalent attachment of streptavidin, FICLI is able to recognize a multilayer formation of the silane and the protein. For immune complexation, the technique is able to detect different antibody-antigen bindings and distinguish different dynamics among different immune species.

3D Nanoporous Anodic Alumina Structures for Sustained Drug Release.

The use of nanoporous anodic alumina (NAA) for the development of drug delivery systems has gained much attention in recent years. The release of drugs loaded inside NAA pores is complex and depends on the morphology of the pores. In this study, NAA, with different three-dimensional (3D) pore structures (cylindrical pores with several pore diameters, multilayered nanofunnels, and multilayered inverted funnels) were fabricated, and their respective drug delivery rates were studied and modeled using doxorubicin as a model drug. The obtained results reveal optimal modeling of all 3D pore structures, differentiating two drug release stages. Thus, an initial short-term and a sustained long-term release were successfully modeled by the Higuchi and the Korsmeyer-Peppas equations, respectively. This study demonstrates the influence of pore geometries on drug release rates, and further presents a sustained long-term drug release that exceeds 60 days without an undesired initial burst.

A Mycoplasma Genomic DNA Probe using Gated Nanoporous Anodic Alumina.

A nanoporous anodic alumina (NAA)-based sensor system for the detection of Mycoplasma was developed through the implementation of "molecular gates" selective to the presence of this bacterium. The capped support showed a negligible cargo release, while presence of Mycoplasma genomic DNA resulted in the release of rhodamine B fluorescent dye. This sensor system presents a limit of detection of 20 genomic DNA copies⋅µL-1 and was applied to the detection of Mycoplasma bacteria in competitive environments, such as culture cell media.

Sustained, Controlled and Stimuli-Responsive Drug Release Systems Based on Nanoporous Anodic Alumina with Layer-by-Layer Polyelectrolyte.

Controlled drug delivery systems are an encouraging solution to some drug disadvantages such as reduced solubility, deprived biodistribution, tissue damage, fast breakdown of the drug, cytotoxicity, or side effects. Self-ordered nanoporous anodic alumina is an auspicious material for drug delivery due to its biocompatibility, stability, and controllable pore geometry. Its use in drug delivery applications has been explored in several fields, including therapeutic devices for bone and dental tissue engineering, coronary stent implants, and carriers for transplanted cells. In this work, we have created and analyzed a stimuli-responsive drug delivery system based on layer-by-layer pH-responsive polyelectrolyte and nanoporous anodic alumina. The results demonstrate that it is possible to control the drug release using a polyelectrolyte multilayer coating that will act as a gate.

Nanoporous Anodic Alumina 3D FDTD Modelling for a Broad Range of Inter-pore Distances.

The capability of the finite difference time domain (FDTD) method for the numerical modelling of the optical properties of nanoporous anodic alumina (NAA) in a broad range of inter-pore distances is evaluated. FDTD permits taking into account in the same numerical framework all the structural features of NAA, such as the texturization of the interfaces or the incorporation of electrolyte anions in the aluminium oxide host. The evaluation is carried out by comparing reflectance measurements from two samples with two very different inter-pore distances with the simulation results. Results show that considering the texturization is crucial to obtain good agreement with the measurements. On the other hand, including the anionic layer in the model leads to a second-order contribution to the reflectance spectrum.

Molecular gated nanoporous anodic alumina for the detection of cocaine.

We present herein the use of nanoporous anodic alumina (NAA) as a suitable support to implement "molecular gates" for sensing applications. In our design, a NAA support is loaded with a fluorescent reporter (rhodamine B) and functionalized with a short single-stranded DNA. Then pores are blocked by the subsequent hybridisation of a specific cocaine aptamer. The response of the gated material was studied in aqueous solution. In a typical experiment, the support was immersed in hybridisation buffer solution in the absence or presence of cocaine. At certain times, the release of rhodamine B from pore voids was measured by fluorescence spectroscopy. The capped NAA support showed poor cargo delivery, but presence of cocaine in the solution selectively induced rhodamine B release. By this simple procedure a limit of detection as low as 5 × 10-7 M was calculated for cocaine. The gated NAA was successfully applied to detect cocaine in saliva samples and the possible re-use of the nanostructures was assessed. Based on these results, we believe that NAA could be a suitable support to prepare optical gated probes with a synergic combination of the favourable features of selected gated sensing systems and NAA.

In Vitro Biocompatibility of Surface-Modified Porous Alumina Particles for HepG2 Tumor Cells: Toward Early Diagnosis and Targeted Treatment.

Porous alumina photoluminescence-inherent particles are produced and proposed for the development of biomarkers detectors and localized treatment of HepG2 cells. Nanoporous alumina particles (NPAPs) are amorphous, consist of hexagonally ordered nanometric pores in an alumina matrix, have high chemical stability in physiological pH, and exhibit a high inherent photoluminescence in the visible spectrum independently of their size, selectable from nanometers to tens of micrometers. The surface of NPAPs is chemically modified using two different functionalization methods, a multistep method with (3-aminopropyl)triethoxysilane (APTES) and glutaraldehyde (GLTA) and a novel simplified-step method with silane-PEG-NHS. Fourier Transform infrared spectroscopy analysis confirmed the proper surface modification of the particles for both functionalization methods. HepG2 cells were cultured during different times with growing concentrations of particles. The analysis of cytotoxicity and cell viability of HepG2 cells confirmed the good biocompatibility of NPAPs in all culture conditions. The results prove the suitability of NPAPs for developing new label-free biomarker detectors and advantageous carriers for localized drug delivery.

Protein attachment to nanoporous anodic alumina for biotechnological applications: influence of pore size, protein size and functionalization path.

Nanoporous anodic alumina (NAA) is a material with great interest in nanotechnology and with promising applications to biotechnology. Obtaining specific and regularly functionalized NAA surfaces is essential to obtain meaningful results and applications. Silane-PEG-NHS (triethoxysilane-polyethylene-glycol-N-hydroxysuccinimide) is a covalent linker commonly used for single-molecule studies. We investigate the functionalization of NAA with silane-PEG-NHS and compared with two common, but not single-molecule, grafting agents, APTMS (3-aminopropylotrimethoxysilane) as an electrostatic linker, and APTMS-GTA (3-aminopropylotrimethoxysilane-glutaraldehyde) as covalent. Another outcome of this study is to show how two proteins (collagen and bovine serum albumin, BSA) with different properties differentially arrange for different functionalizations and NAA pore sizes. FTIR is used to demonstrate the surface modification steps and fluorescence confocal microscopy reveals that silane-PEG-NHS results in a more homogeneous protein distribution in comparison to the other linkers. Reflection interference Fourier transform spectroscopy confirms the confocal fluorescence microscopy results and permits to estimate the amounts of linker and linked proteins within the pores. These results permit to obtain uniformly chemical modified NAA supports with a great value in biosensing, drug delivery and cell biology.

Mirror-image-induced magnetic modes.

Reflection in a mirror changes the handedness of the real world, and right-handed objects turn left-handed and vice versa (M. Gardner, The Ambidextrous Universe, Penguin Books, 1964). Also, we learn from electromagnetism textbooks that a flat metallic mirror transforms an electric charge into a virtual opposite charge. Consequently, the mirror image of a magnet is another parallel virtual magnet as the mirror image changes both the charge sign and the curl handedness. Here we report the dramatic modification in the optical response of a silicon nanocavity induced by the interaction with its image through a flat metallic mirror. The system of real and virtual dipoles can be interpreted as an effective magnetic dipole responsible for a strong enhancement of the cavity scattering cross section.

Porous silicon microcavities: synthesis, characterization, and application to photonic barcode devices.

We have recently developed a new type of porous silicon we name as porous silicon colloids. They consist of almost perfect spherical silicon nanoparticles with a very smooth surface, able to scatter (and also trap) light very efficiently in a large-span frequency range. Porous silicon colloids have unique properties because of the following: (a) they behave as optical microcavities with a high refractive index, and (b) the intrinsic photoluminescence (PL) emission is coupled to the optical modes of the microcavity resulting in a unique luminescence spectrum profile. The PL spectrum constitutes an optical fingerprint identifying each particle, with application for biosensing.In this paper, we review the synthesis of silicon colloids for developing porous nanoparticles. We also report on the optical properties with special emphasis in the PL emission of porous silicon microcavities. Finally, we present the photonic barcode concept.