Organo-soluble porphyrin mixed monolayer-protected gold nanorods with intercalated fullerenes

Organo-soluble porphyrin mixed monolayer-protected gold nanorods were synthesized and characterized. The resulting gold nanorods encapsulated by both porphyrin thiol and alkyl thiol on their entire surface with strong covalent Au-S linkages were very stable in organic solvents without aggregation or decomposition and exhibited unique optical properties different from their corresponding spherical ones. Alkyl thiol acts as a stabilizer not only to fill up the potential space on gold nanorod surface between bulky porphyrin molecules but also to provide space for further insertion of C(60) molecules forming a stable C(60)-porphyrin-gold nanorod hybrid nanostructure.     

Preferential end-to-end assembly of gold nanorods by biotin-streptavidin connectors

A major challenge in nanoscience and nanotechnology is the rational assembly of nanoscale objects. Here we report that gold nanorods, aspect ratio 18, can be functionalized with a biotin disulfide, and subsequent addition of streptavidin links the rods together in an end-to-end manner much more often than expected.     

Oriented assembly of Au nanorods using biorecognition system

The design and formation of a linear assembly of gold nanorods using a biomolecular recognition system are described. Anti-mouse IgG was immobilized on the {111} end faces of gold nanorods through a thioctic acid containing a terminal carboxyl group. The biofunctionalized nanorods can be assembled with the desired length using mouse IgG for biorecognition and binding. The gold nanorods can be assembled to extended nanorod chains, which can be as long as 3 microm. These assembled nanostructures may be used as the precursors for future nanodevices.     

Preparation of biotinylated glyconanoparticles via a photochemical process and study of their bioconjugation to streptavidin

We report here the preparation of novel biotinylated glyconanoparticles from well-defined biotinylated glycopolymers and poly(N-isopropylacrylamide) (PNIPAAm) synthesized via the reversible addition fragmentation chain transfer (RAFT) polymerization process. The in situ reduction of the biotinylated glycopolymers, PNIPAAm, poly(ethylene glycol), and HAuCl4 via a photochemical process resulted in the formation of biotinylated gold nanoparticles. The multifunctional biotinylated glyconanoparticles were then evaluated for their bioconjugation toward streptavidin using UV-vis spectroscopy and surface plasmon resonance (SPR). The biotinylated nanoparticles underwent aggregation in the presence of streptavidin as revealed by spectrophotometry, which indicates the accessibility of the biotin for conjugation. These results were further confirmed by surface plasmon resonance even in the case of surface-immobilized streptavidin.     

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).     

In situ monitoring of the orientated assembly of strep-tagged membrane proteins on the gold surface by surface enhanced infrared absorption spectroscopy

Surface enhanced infrared absorption spectroscopy (SEIRAS) has been employed to monitor the orientated assembly of a strep-tagged membrane protein on the gold surface via a streptavidin/biotin interlayer. The high surface sensitivity of SEIRAS allows for tracking the individual assembling steps on the molecular level. The sequence of surface modification steps comprises: (i) cross-linking of biotin to the self-assembled monolayer of cysteamine along the gold surface; (ii) adsorption of streptavidin to and desorption from the biotin layer; and (iii) adsorption of the strep-tagged membrane protein ecgltP (glutamate transporter of E. coli) on the streptavidin/biotin layer. The analysis of the SEIRA spectra reveals that the biotin layer undergoes a phase transition from an isotropic orientation to a densely packed layer during coupling to the cysteamine monolayer. Formation of the densely packed layer weakens the interaction between streptavidin and the biotin layer but yields a binding specificity of 80%. The specificity of strep-tagged ecgltP to the streptavidin layer is with 60% only modest. Nevertheless, the streptavidin/biotin interlayer reveals a higher regeneration propensity than the His-tag/Ni-NTA interlayer.     

DNA-Mediated Self-Assembly and Metallization of Semiconductor Nanorods for the Fabrication of Nanoelectronic Interfaces

DNA nanostructures provide a powerful platform for the programmable assembly of nanomaterials. Here, this approach is extended to semiconductor nanorods that possess interesting electrical properties and could be utilized for the bottom-up fabrication of nanoelectronic building blocks. The assembly scheme is based on an efficient DNA functionalization of the nanorods. A complete coverage of the rod surface with DNA ensures a high colloidal stability while maintaining the rod size and shape. It furthermore supports the assembly of the nanorods at defined docking positions of a DNA origami platform with binding efficiencies of up to 90 % as well as the formation of nanorod dimers with defined relative orientations. By incorporating orthogonal binding sites for gold nanoparticles, defined metal-semiconductor heterostructures can be fabricated. Subsequent application of a seeded growth procedure onto the gold nanoparticles (AuNPs) allows for to establish a direct metal-semiconductor interface as a crucial basis for the integration of semiconductors in self-assembled nanoelectronic devices.     

Precise placement of gold nanorods by capillary assembly

Capillary assembly was explored for the precise placement of 25 nm × 70 nm colloidal gold nanorods on prestructured poly(dimethylsiloxane) template surfaces. The concentration of nanorods and cationic surfactant cetyltrimethylammonium bromide (CTAB), the template wettability, and most critically the convective transport of the dispersed nanorods were tuned to study their effect on the resulting assembly yield. It is shown that gold nanorods can be placed into arrayed 120-nm diameter holes, achieving assembly yields as high as 95% when the local concentration of nanorods at the receding contact line is sufficiently high. Regular arrays of gold nanorods have several benefits over randomly deposited nanorod arrangements. Each assembled nanorod resides at a precisely defined location and can easily be found for subsequent characterization or direct utilization in a device. The former is illustrated by collecting scattering spectra from single nanorods and nanorod dimers, followed by subsequent SEM characterization without the need for intricate registration schemes.     

One-, two-, and three-dimensional superstructures of gold nanorods induced by dimercaptosuccinic acid

A method is described for assembling gold nanorods into one-, two-, and three-dimensional superstructures. The addition of dimercaptosuccinic acid (DMSA) into the nanorod solution was found to induce self-assembly of the latter to one-dimensional "tapelike", two-dimensional "sheetlike" and three-dimensional "superlattice-like" structures depending on the DMSA concentration. The assembly was found to follow a smectic structure, where the nanorod long axes are parallel to each other. The rods are spaced 8.5 +/- 0.3 nm apart in the resulting structures, which extend over several micrometers in length. Organizations perpendicular to the grid were also found. The nanorod tapes were found to bend, and they form circular assemblies as well. The assembly and morphology of the nanorod structures were characterized by transmission electron microscopy and UV-vis spectroscopy. The effect of the DMSA concentration as well as the pH of the medium was also studied. On the basis of several control experiments utilizing similar molecules, charge neutralization of the nanorods by the carboxylic group of DMSA was found to be the principal reason for such an assembly, while the mercapto groups render additional stability to its structure. A mechanistic model of the assembly is proposed. This type of assembly would plausibly function as a plasmonic waveguide in potential nanodevices.     

Single-molecule detection of DNA via sequence-specific links between F1-ATPase motors and gold nanorod sensors

We report the construction of a novel biosensing nanodevice to detect single, sequence-specific target DNA molecules. Nanodevice assembly occurs through the association of an immobilized F1-ATPase molecular motor and a functionalized gold nanorod via a single 3',5'-dibiotinylated DNA molecule. Target-dependent 3',5'-dibiotinylated DNA bridges form by combining ligation and exonucleation reactions (LXR), with a specificity capable of selecting against a single nucleotide polymorphism (SNP). Using dark field microscopy to detect gold nanorods, quantitation of assembled nanodevices is sufficient to distinguish the presence of as few as 1800 DNA bridges from nonspecifically bound nanorods. The rotary mechanism of F1-ATPase can drive gold nanorod rotation when the nanorod is attached via the DNA bridge. Therefore, rotation discriminates fully assembled devices from nonspecifically bound nanorods, resulting in a sensitivity limit of one zeptomole (600 molecules).     

Highly sensitive biomolecule detection on a quartz crystal microbalance using gold nanoparticles as signal amplification probes.

We report here a novel strategy for the high-sensitive detection of target biomolecules with very low concentrations on a quartz crystal microbalance (QCM) device using gold nanoparticles as signal enhancement probes. By employing a streptavidin-biotin interaction as a model system, we could prepare biotin-conjugated gold nanoparticles maintaining good dispersion and long-term stability by controlling the biotin density on the surface of gold nanoparticles that have been investigated by UV-vis spectra and AFM images. These results showed that 10 microM N-(6-[biotinamido]hexyl)-3'-(2'-pyridyldithio)propionamide (biotin-HPDP) was the critical concentration to prevent the nonspecific aggregation of gold nanoparticles in this system. For sensing streptavidin target molecules by QCM, biotinylated BSA was absorbed on the Au surface of the QCM electrode and subsequent coupling of the target streptavidin to the biotin in the sensing interface followed. Amplification of the sensing process was performed by the interaction of the target streptavidin on the sensing surface with gold nanoparticles modified with 10 microM biotin-HPDP. The biotinylated gold nanoparticles were used as signal amplification probes to improve the detection limit, which was 50 ng/ml, of the streptavidin detection system without signal enhancement, and the calibration curve determined for the net frequency changes showed good linearity over a wide range from 1 ng/ml to 10 microg/ml for the quantitative streptavidin target molecule analysis. In addition, the measured dissipation changes suggested that the layer of biotin-BSA adsorbed on the Au electrode and the streptavidin layer assembled on the biotin-BSA surface were highly compact and rigid. On the other hand, the structure formed by the biotinylated gold nanoparticles on the streptavidin layer was flexible and dissipative, being elongated outward from the sensing surface.     

Plasmon coupling in nanorod assemblies: optical absorption, discrete dipole approximation simulation, and exciton-coupling model

The shape anisotropy of nanorods gives rise to two distinct orientational modes by which nanorods can be assembled, i.e., end-to-end and side-by-side, analogous to the well-known H and J aggregation in organic chromophores. Optical absorption spectra of gold nanorods have earlier been observed to show a red-shift of the longitudinal plasmon band for the end-to-end linkage of nanorods, resulting from the plasmon coupling between neighboring nanoparticles, similar to the assembly of gold nanospheres. We observe, however, that side-by-side linkage of nanorods in solution shows a blue-shift of the longitudinal plasmon band and a red-shift of the transverse plasmon band. Optical spectra calculated using the discrete dipole approximation method were used to simulate plasmon coupling in assembled nanorod dimers. The longitudinal plasmon band is found to shift to lower energies for end-to-end assembly, but a shift to higher energies is found for the side-by-side orientation, in agreement with the optical absorption experiments. The strength of plasmon coupling was seen to increase with decreasing internanorod distance and an increase in the number of interacting nanorods. For both side-by-side and end-to-end assemblies, the strength of the longitudinal plasmon coupling increases with increasing nanorod aspect ratio as a result of the increasing dipole moment of the longitudinal plasmon. For both the side-by-side and end-to-end orientation, the simulation of a dimer of nanorods having dissimilar aspect ratios showed a longitudinal plasmon resonance with both a blue-shifted and a red-shifted component, as a result of symmetry breaking. A similar result is observed for a pair of similar aspect ratio nanorods assembled in a nonparallel orientation. The internanorod plasmon coupling scheme concluded from the experimental results and simulations is found to be qualitatively consistent with the molecular exciton coupling theory, which has been used to describe the optical spectra of H and J aggregates of organic molecules. The coupled nanorod plasmons are also suggested to be electromagnetic analogues of molecular orbitals. Investigation of the plasmon coupling in assembled nanorods is important for the characterization of optical excitations and plasmon propagation in these nanostructures. The surface plasmon resonance shift resulting from nanorod assembly also offers a promising alternative for analyte-sensing assays.     

Programmable Supra‐Assembly of a DNA Surface Adapter for Tunable Chiral Directional Self‐Assembly of Gold Nanorods

An important challenge in molecular assembly and hierarchical molecular engineering is to control and program the directional self‐assembly into chiral structures. Here, we present a versatile DNA surface adapter that can programmably self‐assemble into various chiral supramolecular architectures, thereby regulating the chiral directional “bonding” of gold nanorods decorated by the surface adapter. Distinct optical chirality relevant to the ensemble conformation is demonstrated from the assembled novel stair‐like and coil‐like gold nanorod chiral metastructures, which is strongly affected by the spatial arrangement of neighboring nanorod pair. Our strategy provides new avenues for fabrication of tunable optical metamaterials by manipulating the directional self‐assembly of nanoparticles using programmable surface adapters.     

Chemisorption assembly of Au nanorods on mercaptosilanized glass substrate for label-free nanoplasmon biochip

The fabrication of a localized surface plasmon resonance nanosensor in a chip based format that utilizes Au nanorods (GNRs) as the optical transducer were systematically studied. (3-mercaptopropyl)trimethoxysilane (MPTMS) modified glass substrate offers GNR deposition with maximal sensitivity to local refractive index changes, which subsequently results in better optical recognition of receptor–analyte binding. Kinetics governing the mass transport and chemisorption of nanorods from bulk to solid surface can be dynamically controlled in a predictable fashion. We demonstrate that slight aggregation induced by a low ionic strength (5 mM NaCl) can facilitate the nanorod assembly to result in a dense, well-distributed surface monolayer. In high ionic media (e.g. 40–80 mM), anions present electrostatically bind with the positively charged cetyltrimethylammonium bromide (CTAB) surrounding nanorod surfaces, thereby leading to instability with heavy aggregation in solution. However, once chemically bound on silanized substrates, the nanorods exhibit excellent stability in physiological buffer where high amount of ionic species are present. The fundamental study is followed by demonstration of a practical application of the fabricated biochip in label-free detection based on GNR wavelength shift of the longitudinal palsmon maxima as the optical signature of human IgG model detection.     

Facile preparation of glyconanoparticles and their bioconjugation to streptavidin

Well-defined glycopolymers containing linear and cyclic carbohydrate moieties as pendent groups were prepared by reversible addition fragmentation chain transfer polymerization (RAFT). The RAFT synthesized glycopolymers were used for the aqueous synthesis of stabilized glyconanoparticles. The in situ reduction of the glycopolymers and HAuCl4 resulted in the formation of highly stable modified gold nanoparticles with diameters ranging from 40 to 80 nm in aqueous media. Multifunctional glyconanoparticles were also generated in the presence of varying amounts of biotinylated-polyethyleneglycol (bio-PEG-SH) having terminal thiol groups. The gold nanoparticles underwent aggregation in the presence of streptavidin as revealed by UV-vis spectroscopy. The availability of the biotin for conjugation to streptavidin was also confirmed using surface plasmon resonance (SPR).     

Synthesis of monodisperse biotinylated p(NIPAAm)-coated iron oxide magnetic nanoparticles and their bioconjugation to streptavidin

We describe here the synthesis of 10 nm, monodisperse, iron oxide nanoparticles that we have coated with temperature-sensitive, biotinylated p(NIPAAm) (b-PNIPAAm). The PNIPAAm was prepared by the reversible addition fragmentation chain transfer polymerization (RAFT), and one end was biotinylated with a PEO maleimide-activated biotin to form a stable thioether linkage. The original synthesized iron oxide particles were stabilized with oleic acid. They were dispersed in dioxane, and the oleic acid molecules were then reversibly exchanged with a mixture of PNIPAAm and b-PNIPAAm at 60 degrees C. The b-PNIPAAm-coated magnetic nanoparticles were found to have an average diameter of approximately 15 nm by dynamic light scattering and transmission electron microscopy. The ability of the biotin terminal groups on the b-PNIPAAm-coated nanoparticles to interact with streptavidin was confirmed by fluorescence and surface plasmon resonance. It was found that the b-PNIPAAm-coated iron oxide nanoparticles can still bind with high affinity to streptavidin in solution or when the streptavidin is immobilized on a surface. We have also demonstrated that the binding of the biotin ligands on the surface of the temperature-responsive magnetic nanoparticles to streptavidin can be turned on and off as a function of temperature.     

Steric crowding effects on target detection in an affinity biosensor

This work quantifies the impact of steric crowding on whole antibody (Ab) receptor immobilization and target Ab detection and also demonstrates how the versatile biotin/streptavidin receptor immobilization system must be tuned to optimize target detection in designing biosensors. Results are demonstrated on a label-free optical biosensor fabricated from n-type macroporous porous silicon (PSi) with approximately 88-107 nm diameter pores. We employ a sandwich assay scheme comprising a linking chemistry (biotin/streptavidin) to attach biotinylated anti-rabbit IgG (receptor) to detect rabbit IgG (target). A "bottom-up" approach was taken to investigate each layer of the sandwich assay to optimize target binding. Steric crowding was observed to hinder subsequent layer binding for each layer in the sandwich (biotin, streptavidin, and receptor). Our results give definitive evidence that the onset of steric crowding within the biotin layer occurs at a surface coverage of 57%, which is much higher compared to that from published work on well-ordered self-assembled biotin monolayers on planar gold surfaces. This difference is attributed to the topographical heterogeneity of the PSi substrate. Streptavidin (SA) binding to surface-linked biotin was altered by preblocking the streptavidin binding sites with biotin. Through consistent trends in data, preblocking SA was shown to reduce steric crowding within the SA layer, which translated into increased receptor immobilization. The final detection range of rabbit IgG was 0.07-3 mg mL(-1) (0.4-17 ng mm(-2)), and binding specificity was demonstrated by employing an anti-chicken IgG control receptor. This study underlines the importance of considering binding avidity and surface topography in optimizing chip-based biosensors.     

pH-triggered assembly of gold nanorods

The self-assembly of surfactant-protected gold nanorods (aspect ratio 3.3 +/- 0.3, 20.6 +/- 5.5 nm width, and 67.5 +/- 9.0 nm length) into ordered structures using adipic acid is presented. As made, the gold nanorods are coated with cationic surfactant, which gives them a net positive charge in aqueous solution. The pH-dependent assembly is directed by electrostatic interactions between the positively charged nanorods and negatively charged, deprotonated adipic acid. Absorption spectra and light scattering measurements of these nanorods suggest that aggregation is initiated in solution in the presence of adipic acid at pH 7-8, but not at pH 3, to form small assemblies of nanorods. Zeta potential measurements show that the assembly is significantly less positively charged in the presence of deprotonated adipic acid than when adipic acid is fully protonated.     

Surface-functionalized CdSe nanorods for assembly in diblock copolymer templates

Poly(ethylene oxide)-covered CdSe nanorods were prepared and assembled in diblock copolymer templates by floating the block copolymer templates onto aqueous nanorod solutions. The assembly was enabled by consideration of the surface ligand coverage of the nanorods. Alkane-covered CdSe nanorods prepared by state-of-the-art techniques are not compatible with this assembly process. However, poly(ethylene oxide) (PEO)-functionalized CdSe nanorods were successfully used to assemble the nanorods into the channels and pores of diblock copolymer templates. Other water-dispersible CdSe nanorods, such as those covered with 11-mercaptoundecanoic acid (MUA), did not give the desired assemblies. These results are understood by considering the surface energies of the PEO-covered CdSe nanorods in this interfacial assembly process.     

A calorimetric investigation of the assembly of gold nanorods to form necklaces

Interaction of gold nanorods with cysteine as well as 3-mercaptopropionic acid (MPA) has been investigated by isothermal titration calorimetry (ITC), in combination with electronic absorption spectroscopy and electron microscopy. The assembly process with MPA shows two steps, the first due to the binding of MPA to gold nanorods through the sulfur atom, and the second due to assembly of the MPA capped gold nanorods through the formation of cyclic hydrogen bonded dimers between the MPA molecules. In the case of cysteine only a single step is observed in ITC, due to the binding of the molecules to gold nanorods.