High-capacity and high-intensity DNA microarray spots using oxygen-plasma nanotextured polystyrene slides

Commercially available polystyrene (PS) slides were plasma nanotextured (nano-roughened) through treatment in oxygen plasma discharges to create substrates with increased surface area for microarray applications. Conditions of plasma treatment were determined for maximum and uniform oligonucleotide immobilization on these nanotextured PS slides. Oligonucleotides were immobilized onto the surface in the form of biotinylated oligonucleotide/streptavidin conjugates to take advantage of increased protein binding capacity of the substrate. It was found that the amount of oligonucleotides that could be immobilized was increased up to ten times on plasma treated as compared with untreated slides. The sensitivity of detection of labelled hybridized probes was improved by a factor of 20. Optimized nanotextured PS slides were subsequently used to develop a microarray for the detection of three deleterious BRCA1 gene mutations by immobilizing oligonucleotides corresponding to wild and mutant-type sequences. The microarray developed on the nanotextured PS slides provided higher specific hybridization signal and discrimination ratios as compared with flat untreated PS slides.     

Surface potential mapping of dispersed proteins

We describe a method for detecting proteins after transfer to PVDF membranes, based on the surface potential attributed to each protein. Proteins separated by classical two-dimensional polyacrylamide gel electrophoresis could be detected by scanning the membrane surface with a vibrating capacitor (also called a Kelvin probe) on the basis of differences between their surface potential and that of the membrane. Coupled to colloidal gold staining, the technique enables detection of proteins previously undetectable by classical staining methods. Plotting variations of the surface potential in two dimensions visualizes proteins which migrate close together. Finally, we demonstrate that the Kelvin probe detects proteins over a concentration range from micro to sub-nanogram with increased sensitivity at lower concentrations, and unlike other methods, appears to be similar for all proteins tested so far. The method described is fast, reliable, and it can be automated for high throughput.     

Visible-laser desorption/ionization on gold nanostructures

In this report, we describe the visible-laser desorption/ionization of biomolecules deposited on gold-coated porous silicon and gold nanorod arrays. The porous silicon made by electrochemical etching was coated with gold using argon ion sputtering. The gold nanorod arrays were fabricated by electrodepositing gold onto a porous alumina template, and the subsequent partial removal of the alumina template. A frequency-doubled/tripled Nd : YAG laser was used to irradiate the gold nanostructured substrate, and the desorbed molecular ions were mass-analyzed by a time-of-flight mass spectrometer. The desorption/ionization of biomolecules for both substrates was favored by the use of the 532-nm visible-laser, which is in the range of the localized surface plasmon resonance of the gold nanostructure. The present technique offers a potential analytical method for low-molecular-weight analytes that are rather difficult to handle in the conventional matrix-assisted laser desorption/ionization (MALDI) mass spectrometry.     

Substrate-independent dip-pen nanolithography based on reactive coatings

We report that nanostructuring via dip-pen nanolithography can be used for modification of a broad range of different substrates (polystyrene, Teflon, stainless steel, glass, silicon, rubber, etc.) without the need for reconfiguring the underlying printing technology. This is made possible through the use of vapor-based coatings that can be deposited on these substrates with excellent conformity, while providing functional groups for subsequent spatially directed click chemistry via dip-pen nanolithography. Pattern quality has been compared on six different substrates demonstrating that this approach indeed results in a surface modification protocol with potential use for a wide range of biotechnological applications.     

A reversible microarray immobilization strategy based on thiol-quinone reaction

Microarray technology has been widely applied in biomedical research. The key to microarray study is to develop efficient immobilization method. In this study, we designed a new reversible microarray immobilization method based on thiol-quinone reaction. A quinone-functionalized slide was fabricated through H₂O₂ treatment of dopamine-coated slides. Various thiol-containing molecules can be anchored onto the quinone-functionalized slides via thioether linker, which could be ...     

Theoretical investigation of substrate effect on deliquescence relative humidity of NaCl particles

A theoretical investigation is conducted for the first time to explore the deliquescence of particles deposited on a substrate. The formulation incorporates the Kelvin effect with the assumption that the dry and wet particles are both spherical caps in shape. Unlike the deposited particles larger than 500 nm, the deliquescence relative humidity (DRH) of smaller particles can substantially depend on the particle size, the contact angles, and the surface tension between the particle and the atmosphere. At certain contact angles, small particles depositing on a substrate could deliquesce at a much lower RH, posing a potential corrosion problem for metallic substrates.     

Immobilization of gold nanorods onto acid-terminated self-assembled monolayers via electrostatic interactions

We report the immobilization of gold nanorods onto self-assembled monolayers (SAMs) of 16-mercaptohexadecanoic acid (16-MHA). The simple two step protocol involves formation of a SAM of 16-MHA molecules onto gold-coated glass slides and subsequent immersion of these slides into the gold nanorod solution. The nanorods, formed by a seed-mediated, surfactant-assisted synthesis protocol, are stabilized in solution due to surface modification by the surfactant cetyltrimethylammonium bromide (CTAB). Attractive electrostatic interactions between the carboxylic acid group on the SAM and the positively charged CTAB molecules are likely responsible for the nanorod immobilization. UV-vis spectroscopy has been used to follow the kinetics of the nanorod immobilization. The nature of interaction between the gold nanorods and the 16-MHA SAM has been probed by Fourier transform infrared spectroscopy (FTIR). The surface morphology of the immobilized rods is studied by scanning electron microscopy (SEM) and atomic force microscopy (AFM) measurements. SEM was also used to determine the density of the immobilized nanorods as a function of the pH of immobilization. Control over the surface coverage of the immobilized gold nanorods has been demonstrated by simple pH variation. Such well-dispersed immobilized gold nanorods with control over the surface coverage could be interesting substrates for applications such as surface-enhanced Raman spectroscopy (SERS).     

Protein arrays on high-surface-area plasma-nanotextured poly(dimethylsiloxane)-coated glass slides

Treatment of poly(dimethylsiloxane) (PDMS) surfaces with SF(6) plasma results in the creation of high-surface-area nanotextured surfaces that considerably favour protein adsorption with respect to untreated ones. In order to employ such nanotextured surfaces as substrates for microarrays to be created and analysed using standard instrumentation, we fabricated thin PDMS films on top of standard low-cost microscope glass slides. The properties of both untreated and plasma-treated PDMS-coated slides towards spotting of protein solutions were evaluated in terms of spot signal intensity and homogeneity as well as of spot shape and size. It was found that the plasma-treated PDMS-coated glass slides provided highly homogeneous spots (mean intra-spot variation 7.6%) with spot signal intensity 6-times higher than that obtained using the untreated ones. In addition, comparison with commercially available polystyrene and aminosilanized-glass microarray slides showed that the proposed slides provided 3-times higher spot signal intensity and 2-times lower intra-spot signal variation. In addition, the implementation of long-aged-after-plasma-treatment nanotextured PDMS-coated glass slides provided spots whose shape and size matched those of the spotting tip. As a consequence, denser arrays of variable spot shape can be created using SF(6) plasma-treated PDMS-coated slides instead of standard microarray slides opening new potentials for bioanalytical applications.     

Developments in the theoretical modelling and experimental measurement of the surface potential of condensed monolayers

In recent years considerable progress has been made in developing a theory for the surface potential of monolayers both at the air-water interface and deposited onto solid supports. This period has also seen the advent of scanning probe technology which has enabled surface potential to be measured to a hitherto undreamed of spatial resolution. This paper traces the key stages in these developments and explores the challenges that remain. Initially, the various models proposed for relating molecular and bond dipole moments to the surface potential are evaluated and the reliability of the moments obtained by applying the models to experimental results investigated. The limitations of the traditional Kelvin probe method of measuring surface potential are then highlighted and how these are overcome in the new generation of scanning force microscopes. Finally, it is suggested that such instruments could readily form the basis of a 'READ' head for a molecular memory based on a self-assembled, macromolecular lattice.     

Fluorous-based peptide microarrays for protease screening

As ever more protease sequences are uncovered through genome sequencing projects, efficient parallel methods to discover the potential substrates of these proteases becomes crucial. Herein we describe the first use of fluorous-based microarrays to probe peptide sequences and begin to define the scope and limitations of fluorous microarray technologies for the screening of proteases. Comparison of a series of serine proteases showed that their ability to cleave peptide substrates in solution was maintained upon immobilization of these substrates onto fluorous-coated glass slides. The fluorous surface did not serve to significantly inactivate the enzymes. However, addition of hydrophilic components to the peptide sequences could induce lower rates of substrate cleavage with enzymes such as chymotrypsin with affinities to hydrophobic moieties. This work represents the first step to creating robust protease screening platforms using noncovalent microarray interface that can easily incorporate a range of compounds on the same slide.     

Corrigendum to “Deposition of chemically reactive and repellent sites on biosensor chips for reduced non-specific binding” [Colloids Surf. B 79 (2010) 270–275]

Treatment of poly(dimethylsiloxane) (PDMS) surfaces with SF₆ plasma results in the creation of high-surface-area nanotextured surfaces that considerably favour protein adsorption with respect to untreated ones. In order to employ such nanotextured surfaces as substrates for microarrays to be created and analysed using standard instrumentation, we fabricated thin PDMS films on top of standard low-cost microscope glass slides. The properties of both untreated and plasma-treated PDMS-coated slides towards spotting of protein solutions were evaluated in terms of spot signal intensity and homogeneity as well as of spot shape and size. It was found that the plasma-treated PDMS-coated glass slides provided highly homogeneous spots (mean intra-spot variation 7.6%) with spot signal intensity 6-times higher than that obtained using the untreated ones. In addition, comparison with commercially available polystyrene and aminosilanized-glass microarray slides showed that the proposed slides provided 3-times higher spot signal intensity and 2-times lower intra-spot signal variation. In addition, the implementation of long-aged-after-plasma-treatment nanotextured PDMS-coated glass slides provided spots whose shape and size matched those of the spotting tip. As a consequence, denser arrays of variable spot shape can be created using SF₆ plasma-treated PDMS-coated slides instead of standard microarray slides opening new potentials for bioanalytical applications.     

Matrix‐assisted laser ablation production of gold cluster ions from Au‐coated photonic crystals

A new strategy was explored to generate pure gold cluster ions, Au_n^+/?, from gold films deposited on solid substrates via a matrix‐assisted laser ablation technique. The gold films deposited on SiO₂‐particle‐assembled photonic crystals were demonstrated to be the most ideal compared with the films deposited on various glass slides. Dropped with a matrix of 2‐(4‐hydroxyphenylazo) benzoic acid and bombarded by nitrogen pulse laser (355 nm), they could release a series of Au_n⁺ with n more than 110 or Au_n^? with n more than 60 according to the data obtained by inline time‐of‐flight mass spectrometry. The gold‐deposited photonic crystal substrates could be stored at room temperature for at least 6 months. The method is hence steady and convenient in use. Copyright © 2012 John Wiley & Sons, Ltd.     

C_(60)离子束撞击固体表面的坍塌沉积(Ⅰ)共焦显微拉曼光谱研究

为了研究C60的结构特性，我们最近在实验中将加这后的C60离子束沉积在固体表面，对其沉积后的形态进行了共焦显微拉曼光谱的表征．在记录的光谱中已检测不出C60原有的特征谱问，说明C60离子在高速憧击固体表面后，已经完全失去了原有的球状构型．C60的t）［R实验在自制的串级飞     

C60离子束撞击固体表面的坍塌沉积(I)共焦显微拉曼光谱研究

Raman spectroscopy was applied to characterize the species deposited from the mass-selected C60 ion beam which was accelerated to 900 eV. The substrates for the deposition were (0001) surface of highly oriented pyrolitic graphite and (111) surface of gold crystal. The species do not exhibit the Raman scattering features of buckminsterfullerene, but displays peaks at 1585 and 1332 cm-1 instead. The former peak is the chararteristic Ranan peak of hexagonal graphite, and the latter can be attributed to the amorphous carbon in sp3 hybridization. The result reveals that C60 was collapsed to form a new phase when it collides on the solid surface.     

Fabrication of DNA microarrays on nanoengineered polymeric ultrathin film prepared by self-assembly of polyelectrolyte multilayers

Microarray-based technology is in need of flexible and cost-effective chemistry for fabrication of oligonucleotide microarrays. We have developed a novel method for the fabrication of oligonucleotide microarrays with unmodified oligonucleotide probes on nanoengineered three-dimensional thin films that are deposited on glass slides by consecutive layer-to-layer adsorption of polyelectrolytes. Unmodified oligonucleotide probes were spotted and immobilized on these multilayered polyelectrolyte thin films (PET) by electrostatic adsorption and entrapment on the porous structure of the PET film. The PET provides higher probe binding capacity and thus higher hybridization signal than that of the traditional two-dimensional aminosilane and poly-L-lysine coated slides. Immobilized probe densities of 3.4 x 10(12)/cm2 were observed for microarray spots on PET with unmodified 50-mer oligonucleotide probes, which is comparable to the immobilized probe densities of alkyamine-modified 50-mer probes end-tethered on an aldehyde-functionalized slide. The study of hybridization efficiency showed that 90% of immobilized probes on PET film are accessible to target DNA to form duplex format in hybridization. The DNA microarray fabricated on PET film has wider dynamic range (about 3 orders of magnitude) and lower detection limit (0.5 nM) than the conventional amino- and aldehyde-functionalized slides. Oligonucleotide microarrays fabricated on these PET-coated slides also had consistent spot morphology. In addition, discrimination of single nucleotide polymorphism of 16S rRNA genes was achieved with the PET-based oligonucleotide microarrays. The PET microarrays constructed by our self-assembly process is cost-effective, versatile, and well suited for immobilizing many types of biological active molecules so that a wide variety of microarray formats can be developed.     

Surface-enhanced Raman scattering: realization of localized surface plasmon resonance using unique substrates and methods

Surface-enhanced Raman scattering (SERS) enhancement and the reproducibility of the SERS signal strongly reflect the quality and nature of the SERS substrates because of diverse localized surface plasmon resonance (LSPR) excitations excited at interstitials or sharp edges. LSPR excitations are the most important ingredients for achieving huge enhancements in the SERS process. In this report, we introduce several gold and silver nanoparticle-based SERS-active substrates developed solely by us and use these substrates to investigate the influence of LSPR excitations on SERS. SERS-active gold substrates were fabricated by immobilizing colloidal gold nanoparticles on glass slides without using any surfactants or electrolytes, whereas most of the SERS-active substrates that use colloidal gold/silver nanoparticles are not free of surfactant. Isolated aggregates, chain-like elongated aggregates and two-dimensional (2D) nanostructures were found to consist mostly of monolayers rather than agglomerations. With reference to correlated LSPR and SERS, combined experiments were carried out on a single platform at the same spatial position. The isolated aggregates mostly show a broadened and shifted SPR peak, whereas a weak blue-shifted peak is observed near 430 nm in addition to broadened peaks centered at 635 and 720 nm in the red spectral region in the chain-like elongated aggregates. In the case of 2D nanostructures, several SPR peaks are observed in diverse frequency regions. The characteristics of LSPR and SERS for the same gold nanoaggregates lead to a good correlation between SPR and SERS images. The elongated gold nanostructures show a higher enhancement of the Raman signal than the the isolated and 2D samples. In the case of SERS-active silver substrates for protein detection, a new approach has been adopted, in contrast to the conventional fabrication method. Colloidal silver nanoparticles are immobilized on the protein functionalized glass slides, and further SERS measurements are carried out based on LSPR excitations. A new strategy for the detection of biomolecules, particularly glutathione, under aqueous conditions is proposed. Finally, supramolecular J-aggregates of ionic dyes incorporated with silver colloidal aggregates are characterized by SERS measurements and correlated to finite-difference time-domain analysis with reference to LSPR excitations. Figure SPR and SERS images for isolated, elongated and two-dimensional gold nanostructures     

Surface immobilized biochemical macromolecules studied by scanning Kelvin microprobe

The measurement of work function is a particularly effective method for the characterization of surfaces because of the sensitivity of the parameter to interfacial structure, modification and overall chemistry. Accordingly, techniques for the analysis of work function offer a powerful tool for monitoring surface chemical changes, especially for situations involving the immobilization of new moieties at the interface. In the present paper, we describe the performance of a new, modified scanning Kelvin microprobe which is capable of the tandem measurement of contact potential and surface topography with resolutions of 1 mV and 10 nm, respectively. The lateral resolution is 1 micron. The instrument has been applied to the study of substrates modified by the attachment of biochemical macromolecules such as oligonucleotides and DNA. This preliminary work confirms the great potential of the technique in the study of biocompatibility, macromolecular structure and microarray devices.     

Defining rules for the shape evolution of gold nanoparticles

The roles of silver ions and halides (chloride, bromide, and iodide) in the seed-mediated synthesis of gold nanostructures have been investigated, and their influence on the growth of 10 classes of nanoparticles that differ in shape has been determined. We systematically studied the effects that each chemical component has on the particle shape, on the rate of particle formation, and on the chemical composition of the particle surface. We demonstrate that halides can be used to (1) adjust the reduction potential of the gold ion species in solution and (2) passivate the gold nanoparticle surface, both of which control the reaction kinetics and thus enable the selective synthesis of a series of different particle shapes. We also show that silver ions can be used as an underpotential deposition agent to access a different set of particle shapes by controlling growth of the resulting gold nanoparticles through surface passivation (more so than kinetic effects). Importantly, we show that the density of silver coverage can be controlled by the amount and type of halide present in solution. This behavior arises from the decreasing stability of the underpotentially deposited silver layer in the presence of larger halides due to the relative strengths of the Ag(+)/Ag(0)-halide and Au(+)/Au(0)-halide interactions, as well as the passivation effects of the halides on the gold particle surface. We summarize this work by proposing a set of design considerations for controlling the growth and final shape of gold nanoparticles prepared by seed-mediated syntheses through the judicious use of halides and silver ions.     

Porous GaN as a template to produce surface-enhanced Raman scattering-active surfaces

Surface-enhanced Raman spectroscopy (SERS) substrates have been prepared by depositing Au or Ag on porous GaN (PGaN). The PGaN used as the template for the metal deposition in these studies was generated by a Pt-assisted electroless etching technique. PGaN was chosen as a potential SERS template due to its nanostructured surface and high surface area, two characteristics that are important for SERS substrates. Metal films were deposited either by solution-based electroless deposition or by thermal vacuum evaporation. SERS spectra were recorded at lambda = 752.5 nm for Au films and at lambda = 514.5 nm for Ag films deposited on PGaN. The SERS signal strength across the metal coated PGaN substrates was uniform and was not plagued by "hot" or "cold" spots on the surface, a common problem with other SERS surfaces. The Ag film deposited by electroless deposition had the highest overall SERS response, with an enhancement factor (EF) relative to normal Raman spectroscopy of 10(8). A portion of the increase in EF relative to typical SERS-active substrates can be assigned to the large surface area characteristic of the PGaN-Ag structures, but some of the enhancement is intrinsic and is likely related to the specific morphology of the metal-nanopore composite structure.