Nanopore blockade sensors for ultrasensitive detection of proteins in complex biological samples.

Nanopore sensors detect individual species passing through a nanoscale pore. This experimental paradigm suffers from long analysis times at low analyte concentration and non-specific signals in complex media. These limit effectiveness of nanopore sensors for quantitative analysis. Here, we address these challenges using antibody-modified magnetic nanoparticles ((anti-PSA)-MNPs) that diffuse at zero magnetic field to capture the analyte, prostate-specific antigen (PSA). The (anti-PSA)-MNPs are magnetically driven to block an array of nanopores rather than translocate through the nanopore. Specificity is obtained by modifying nanopores with anti-PSA antibodies such that PSA molecules captured by (anti-PSA)-MNPs form an immunosandwich in the nanopore. Reversing the magnetic field removes (anti-PSA)-MNPs that have not captured PSA, limiting non-specific effects. The combined features allow detecting PSA in whole blood with a 0.8 fM detection limit. Our 'magnetic nanoparticle, nanopore blockade' concept points towards a strategy to improving nanopore biosensors for quantitative analysis of various protein and nucleic acid species.

Monolayer surface chemistry enables 2-colour single molecule localisation microscopy of adhesive ligands and adhesion proteins.

Nanofabricated and nanopatterned surfaces have revealed the sensitivity of cell adhesion to nanoscale variations in the spacing of adhesive ligands such as the tripeptide arginine-glycine-aspartic acid (RGD). To date, surface characterisation and cell adhesion are often examined in two separate experiments so that the localisation of ligands and adhesion proteins cannot be combined in the same image. Here we developed self-assembled monolayer chemistry for indium tin oxide (ITO) surfaces for single molecule localisation microscopy (SMLM). Cell adhesion and spreading were sensitive to average RGD spacing. At low average RGD spacing, a threshold exists of 0.8 RGD peptides per µm2 that tether cells to the substratum but this does not enable formation of focal adhesions. These findings suggest that cells can sense and engage single adhesive ligands but ligand clustering is required for cell spreading. Thus, our data reveal subtle differences in adhesion biology that may be obscured in ensemble measurements.

A rapid readout for many single plasmonic nanoparticles using dark-field microscopy and digital color analysis.

The integration of plasmonic nanoparticles into biosensors has the potential to increase the sensitivity and dynamic range of detection, through the use of single nanoparticle assays. The analysis of the localized surface plasmon resonance (LSPR) of plasmonic nanoparticles has allowed the limit of detection of biosensors to move towards single molecules. However, due to complex equipment or slow analysis times, these technologies have not been implemented for point-of-care detection. Herein, we demonstrate an advancement in LSPR analysis by presenting a technique, which utilizes an inexpensive CMOS-equipped digital camera and a dark-field microscope, that can analyse the λmax of over several thousand gold nanospheres in less than a second, without the use of a spectrometer. This improves the throughput of single particle spectral analysis by enabling more nanoparticles to be probed and in a much shorter time. This technique has been demonstrated through the detection of interleukin-6 through a core-satellite binding assay. We anticipate that this technique will aid in the development of high-throughput, multiplexed and point-of-care single nanoparticle biosensors.

Thermoresponsive plasmonic core-satellite nanostructures with reversible, temperature sensitive optical properties.

Herein is described a facile method for the assembly of plasmonic gold nanoparticles into smart plasmonic core-satellite nanostructures that allow for the dynamic and reversible tuning of the localised surface plasmon resonance using temperature. This smart system takes advantage of the thermoresponsive polymer linker that modulates the gap distance between the core and satellites in response to the temperature, resulting in the tuning of the surface plasmon coupling and resultant optical shift. It permits optical shifts over a wide wavelength range and reversible control of the optical properties by altering the temperature, which may allow these systems to become candidates for temperature sensitive nanosensors.

Simple Method for Tuning the Optical Properties of Thermoresponsive Plasmonic Nanogels.

We report a straightforward way for forming and tuning the optical properties of thermally responsive plasmonic nanogels. Upon functionalization, a small red shift (2-3 nm) of the pNIPAM@AuNPs was observed due to changes in the refractive index surrounding the AuNP. By adding thermoresponsive poly-N-isopropylacrylamide (pNIPAM) into the pNIPAM@AuNP, its optical response was significantly increased. Heating the nanogel such that the pNIPAM collapsed and acted as a cross-link resulted in the aggregation of the AuNPs. The plasmonic response with red shifts of up to 20 nm was observed. The enlarged red shift was due to the increase in the dielectric constant around the particles and the interparticle interaction of the AuNPs. The interparticle interaction also leads to the broadening of the spectra. Experimental data and finite-difference time-domain (FDTD) calculation are in agreement with this observation. The temperature-dependent optical properties were reversible through multiple cycles of heating and cooling.

Single nanoparticle plasmonic sensors.

The adoption of plasmonic nanomaterials in optical sensors, coupled with the advances in detection techniques, has opened the way for biosensing with single plasmonic particles. Single nanoparticle sensors offer the potential to analyse biochemical interactions at a single-molecule level, thereby allowing us to capture even more information than ensemble measurements. We introduce the concepts behind single nanoparticle sensing and how the localised surface plasmon resonances of these nanoparticles are dependent upon their materials, shape and size. Then we outline the different synthetic approaches, like citrate reduction, seed-mediated and seedless growth, that enable the synthesis of gold and silver nanospheres, nanorods, nanostars, nanoprisms and other nanostructures with tunable sizes. Further, we go into the aspects related to purification and functionalisation of nanoparticles, prior to the fabrication of sensing surfaces. Finally, the recent developments in single nanoparticle detection, spectroscopy and sensing applications are discussed.

Biointerfaces on indium-tin oxide prepared from organophosphonic acid self-assembled monolayers.

Herein we show the development of biointerfaces on indium-tin oxide (ITO) surfaces prepared from organophosphonate self-assembled monolayers. The interfaces were prepared in a stepwise fabrication procedure containing a base monolayer modified with oligo(ethylene oxide) species to which biological recognition ligands were attached. The density of ligands was controlled by varying the ratio of two oligo(ethylene oxide) species such that only one is compatible with further coupling. The final biointerface on ITO was assessed using cell adhesion studies, which showed that the biointerfaces prepared on ITO performed similarly to equivalent monolayers on gold or silicon.

Eutectic liquid alloys for plasmonics: theory and experiment.

We report a method based on density functional theory molecular dynamics that allows us to calculate the plasmonic properties of liquid metals and metal alloys from first principles with no a priori knowledge of the system. We show exceptional agreement between the simulated and measured optical constants of liquid Ga and the room temperature liquid In-Ga eutectic alloy (T(m) = 289 K). We then use this method to analyze the plasmonic properties of various alloy concentrations in the In-Ga system. The plasmonic performance of the In-Ga system decreases with increasing In concentration. However, the benefits of a room-temperature plasmonic liquid are likely to outweigh the minor reduction in plasmonic performance when moving from pure Ga to the eutectic composition. Our results show that density functional theory molecular dynamics can be used as a predictive tool for studying the optical properties of liquid metal systems amenable to plasmonics.

Liquid plasmonics: manipulating surface plasmon polaritons via phase transitions.

This paper reports the manipulation of surface plasmon polaritons (SPPs) in a liquid plasmonic metal by changing its physical phase. Dynamic properties were controlled by solid-to-liquid phase transitions in 1D Ga gratings that were fabricated using a simple molding process. Solid and liquid phases were found to exhibit different plasmonic properties, where light coupled to SPPs more efficiently in the liquid phase. We exploited the supercooling characteristics of Ga to access plasmonic properties associated with the liquid phase over a wider temperature range (up to 30 °C below the melting point of bulk Ga). Ab initio density functional theory-molecular dynamic calculations showed that the broadening of the solid-state electronic band structure was responsible for the superior plasmonic properties of the liquid metal.

Synthesis of inorganic nanomaterials.

Synthesis forms a vital aspect of the science of nanomaterials. In this context, chemical methods have proved to be more effective and versatile than physical methods and have therefore, been employed widely to synthesize a variety of nanomaterials, including zero-dimensional nanocrystals, one-dimensional nanowires and nanotubes as well as two-dimensional nanofilms and nanowalls. Chemical synthesis of inorganic nanomaterials has been pursued vigorously in the last few years and in this article we provide a perspective on the present status of the subject. The article includes a discussion of nanocrystals and nanowires of metals, oxides, chalcogenides and pnictides. In addition, inorganic nanotubes and nanowalls have been reviewed. Some aspects of core-shell particles, oriented attachment and the use of liquid-liquid interfaces are also presented.

Novel vapor phase reactions for the synthesis and modification of carbon nanotubes and inorganic nanowires.

Several vapor phase methods have been developed for the preparation and modification of carbon nanotubes and inorganic nanowires. Thus, nebulized spray pyrolysis has been employed for the synthesis of carbon nanotubes and metal nanowires. Multi-walled carbon nanotubes (MWNTs) with fairly uniform diameters and aligned nanotube bundles have been obtained by nebulized spray pyrolysis using solutions of organometallics such as ferrocene in hydrocarbon solvents. Single-crystalline nanowires of zinc, cadmium, cobalt, and lead are obtained by the decomposition of metal acetates. By reacting acid-treated carbon nanotubes with vapors of metal halides, followed by reaction with water and calcination chemically-bonded oxide layers can be obtained on the nanotubes. A similar procedure has been employed to prepare chemically-bonded oxide layers on Al2O3, ZnO, and silicon nanowires by the reaction of the metal halides with the surface hydroxyl groups present on these nanowire surfaces.

GaS and GaSe nanowalls and their transformation to Ga2O3 and GaN nanowalls.

Two-dimensional nanowalls of GaS and GaSe are obtained by thermal exfoliation around 900 degrees C, and transformed to Ga2O3 and GaN nanowalls upon reaction with air and ammonia respectively at 800 degrees C, while maintaining dimensional integrity.

Generation of onions and nanotubes of GaS and GaSe through laser and thermally induced exfoliation.

Although theoretical calculations have predicted that layered GaS and GaSe should form nanotube and related structures, they have not been prepared experimentally to date. We have carried out laser irradiation as well as thermal treatment of GaS and GaSe to cause exfoliation, which yielded onionlike structures and nanotubes by the rolling of the exfoliated sheets.