Matt Trau

Field-Induced Layering of Colloidal Crystals

An electrohydrodynamic methodology has been developed that makes possible the precise assembly of two- and three-dimensional colloidal crystals on electrode surfaces. Electrophoretically deposited colloidal particles were observed to move toward one another over very large distances (greater than five particle diameters) to form two-dimensional colloidal crystals for both micrometer- and nanometer-size particles. This coalescence of particles with the same charge is opposite to what is expected from electrostatic considerations and appears to result from electrohydrodynamic fluid flow arising from an ionic current flowing through the solution. The ability to modulate this “lateral attraction” between particles, by adjusting field strength or frequency, facilitates the reversible formation of two-dimensional fluid and crystalline colloidal states on the electrode surface. Further manipulation allows controlled structures to be assembled.

Microscopic patterning of orientated mesoscopic silica through guided growth

The supramolecular assembly of surfactant molecules at a solid–liquid interface can produce tubular structures with diameters of around 10 nm (refs 1,2,3,4), which can be used for the templated polymerization of mesoporous silica thin films. The orientation of the tubules depends primarily on the nature of the substrate–surfactant interaction. These nanostructured films hold much promise for applications such as their use as orientated nanowires, sensor/actuator arrays and optoelectronic devices. But a method of patterning the tubules and orientating them into designed arrangements is required for many of these possibilities to be realized. Here we describe a method that allows the direction of growth of these tubules to be guided by infiltrating a reaction fluid into the microcapillaries of a mould in contact with a substrate. An electric field applied tangentially to the surface within the capillaries induces electro-osmotic flow, and also enhances the rates of silica polymerization around the tubules by localized Joule heating. After removal of the mould, patterned bundles of orientated nanotubules remain on the surface. This method permits the formation of orientated mesoporous channels on a non-conducting substrate with an arbitrary microscopic pattern.

Isothermal Detection of DNA by Beacon-Assisted Detection Amplification†

(Figure Presented) Not bad at all: A simple biological circuit composed of two integrated molecular switches was developed for the detection and amplification of a specific nucleic acid in a biological extract. The nucleic acid is recognized by a molecular beacon, then a DNA polymerase and nicking endonuclease are used to rapidly amplify the signal for sensitive detection of the nucleic acid.

Analysis of exosome purification methods using a model liposome system and tunable-resistive pulse sensing

Exosomes are vesicles which have garnered interest due to their diagnostic and therapeutic potential. Isolation of pure yields of exosomes from complex biological fluids whilst preserving their physical characteristics is critical for downstream applications. In this study, we use 100 nm-liposomes from 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and cholesterol as a model system as a model system to assess the effect of exosome isolation protocols on vesicle recovery and size distribution using a single-particle analysis method. We demonstrate that liposome size distribution and ζ-potential are comparable to extracted exosomes, making them an ideal model for comparison studies. Four different purification protocols were evaluated, with liposomes robustly isolated by three of them. Recovered yields varied and liposome size distribution was unaltered during processing, suggesting that these protocols do not induce particle aggregation. This leads us to conclude that the size distribution profile and characteristics of vesicles are stably maintained during processing and purification, suggesting that reports detailing how exosomes derived from tumour cells differ in size to those from normal cells are reporting a real phenomenon. However, we hypothesize that larger particles present in most purified exosome samples represent co-purified contaminating non-exosome debris. These isolation techniques are therefore likely nonspecific and may co-isolate non-exosome material of similar physical properties.

Tunable nano/micropores for particle detection and discrimination: Scanning ion occlusion spectroscopy

Conventional pore technologies are limited in the size range of structures they can analyze by the fixed size of the pore. A novel stretchable pore which can be tuned to optimize the pore size to a particular experimental system is applied here to distinguish between nanoparticle populations of similar size and to detect DNA modification of nanoparticles. Copyright

The fabrication and characterization of biodegradable HA/PHBV nanoparticle-polymer composite scaffolds

This study reports the fabrication and characterization of nano-sized hydroxyapatite (HA)/poly(hydroxyabutyrate-co-hydroxyvalerate) (PHBV) polymer composite scaffolds with high porosity and controlled pore architectures. These scaffolds were prepared using a modified thermally induced phase-separation technique. This investigation focuses on the effect of fabrication conditions on the overall pore architecture of the scaffolds and the dispersion of HA nanocrystals within the composite scaffolds. The morphologies, mechanical properties and in vitro bioactivity of the composite scaffolds were investigated. It was noted that the pore architectures could be manipulated by varying phase-separation parameters. The HA particles were dispersed in the pore walls of the scaffolds and were well bonded to the polymer. The introduction of HA greatly increased the stiffness and strength, and improved the in vitro bioactivity of the scaffolds. The results suggest these newly developed nano-HA/PHBV composite scaffolds may serve as an effective three-dimensional substrate in bone tissue engineering.

Use of tunable nanopore blockade rates to investigate colloidal dispersions

Tunable nanopores fabricated in elastomeric membranes have been used to study the dependence of ionic current blockade rate on the concentration and electrophoretic mobility of particles in aqueous suspensions. A range of nanoparticle sizes, materials and surface functionalities has been tested. Using pressure-driven flow through a pore, the blockade rate for 100 nm carboxylated polystyrene particles was found to be linearly proportional to both transmembrane pressure (between 0 and 1.8 kPa) and particle concentration (between 7 × 10(8) and 4.5 × 10(10) ml( - 1)). This result can be accurately modelled using Nernst-Planck transport theory, enabling measurement of particle concentrations. Using only an applied potential across a pore, the blockade rates for carboxylic acid and amine coated 500 and 200 nm silica particles were found to correspond to changes in their mobility as a function of the solution pH. Scanning electron microscopy and confocal microscopy have been used to visualize changes in the tunable nanopore geometry in three dimensions as a function of applied mechanical strain. The pores were conical in shape, and changes in pore size were consistent with ionic current measurements. A zone of inelastic deformation adjacent to the pore has been identified as important in the tuning process.

Novel miniaturized systems in high-throughput screening.

High-throughput screening (HTS) using high-density microplates is the primary method for the discovery of novel lead candidate molecules. However, new strategies that eschew 2D microplate technology, including technologies that enable mass screening of targets against large combinatorial libraries, have the potential to greatly increase throughput and decrease unit cost. This review presents an overview of state-of-the-art microplate-based HTS technology and includes a discussion of emerging miniaturized systems for HTS. We focus on new methods of encoding combinatorial libraries that promise throughputs of as many as 100,000 compounds per second.

Detecting Exosomes Specifically: A Multiplexed Device Based on Alternating Current Electrohydrodynamic Induced Nanoshearing

Exosomes show promise as noninvasive biomarkers for cancer, but their effective capture and specific detection is a significant challenge. Herein, we report a multiplexed microfluidic device for highly specific capture and detection of multiple exosome targets using a tunable alternating current electrohydrodynamic (ac-EHD) methodology, referred to as nanoshearing. In our system, electrical body forces generated by ac-EHD act within nanometers of an electrode surface (i.e., within the electrical layer) to generate nanoscaled fluid flow that enhances the specificity of capture and also reduce nonspecific adsorption of weakly bound molecules from the electrode surface. This approach demonstrates the analysis of exosomes derived from cells expressing human epidermal growth factor receptor 2 (HER2) and prostate specific antigen (PSA), and is also capable of specifically isolating exosomes from breast cancer patient samples. The device also exhibited a 3-fold enhancement in detection sensitivity in comparison to hydrodynamic flow based assays (LOD 2760 exosomes/μL for ac-EHD vs LOD 8300 exosomes/μL for hydrodynamic flow; (n = 3)). We propose this approach can potentially have relevance as a simple and rapid quantification tool to analyze exosome targets in biological applications.

Methylome sequencing in triple-negative breast cancer reveals distinct methylation clusters with prognostic value

Epigenetic alterations in the cancer methylome are common in breast cancer and provide novel options for tumour stratification. Here, we perform whole-genome methylation capture sequencing on small amounts of DNA isolated from formalin-fixed, paraffin-embedded tissue from triple-negative breast cancer (TNBC) and matched normal samples. We identify differentially methylated regions (DMRs) enriched with promoters associated with transcription factor binding sites and DNA hypersensitive sites. Importantly, we stratify TNBCs into three distinct methylation clusters associated with better or worse prognosis and identify 17 DMRs that show a strong association with overall survival, including DMRs located in the Wilms tumour 1 (WT1) gene, bi-directional-promoter and antisense WT1-AS. Our data reveal that coordinated hypermethylation can occur in oestrogen receptor-negative disease, and that characterizing the epigenetic framework provides a potential signature to stratify TNBCs. Together, our findings demonstrate the feasibility of profiling the cancer methylome with limited archival tissue to identify regulatory regions associated with cancer.