Monitoring of drinking water quality using automated ATP quantification.

A microfluidic based system was developed for automated online method for the rapid detection and monitoring of drinking water contamination utilising microbial Adrenosine-5'-Triphosphate (ATP) as a bacterial indicator. The system comprises a polymethyl methacrylate based microfluidic cartridge inserted into an enclosure incorporating the functions of fluid storage and delivery, lysis steps and real-time detection. Design, integration and operation of the resulting automated system are reported, including the lysis method, the design of the mixing circuit, the choices of flow rate, temperature and reagent amount. Calibration curves of both total and free ATP were demonstrated to be highly linear over a range from 2.5-5000 pg/mL with the limit of detection being lower than 2.5 pg/mL of total ATP. The system was trialled in a lab study with different types of water, with lysis efficiency being found to be strongly dependent upon water type. Further development is required before online implementation.

Photolithographic nanoseeding method for selective synthesis of metal-catalysed nanostructures.

In this work we present a general method for the selective synthesis by photolithography of localised nanostructures in planar geometries. The methodology relies on the previous concept of photo-patternable metallic nanoparticle (NP)/polymer nanocomposites, which can provide a range of NP sizes, polydispersity and densities. First, a photoresist containing metallic ions is patterned by photolithography. Silver NPs are synthesised in situ after the exposure and development of the patterned thin film via the thermal-induced reduction of ions embedded in its structure. Gentle plasma ashing is used to selectively remove the polymer, which leaves NPs on the patterned areas. These NPs are used as seeds for subsequent processes. In order to demonstrate the flexibility of the method, its use to selectively produce localised nanostructures through different processes is shown here. Following a top-down approach, high aspect-ratio silicon nanograss has been produced by reactive ion etching and masking by the NPs. In a bottom-up approach, 280 nm copper clusters have been selectively grown in arrays. This method can be easily extrapolated to other metals and it provides a quick way to selectively generate hierarchical nanostructures in large planar areas that can be used for different applications, such as the fabrication of nanostructured sensor arrays.

Towards enhanced automated elution systems for waterborne protozoa using megasonic energy.

Continuous and reliable monitoring of water sources for human consumption is imperative for public health. For protozoa, which cannot be multiplied efficiently in laboratory settings, concentration and recovery steps are key to a successful detection procedure. Recently, the use of megasonic energy was demonstrated to recover Cryptosporidium from commonly used water industry filtration procedures, forming thereby a basis for a simplified and cost effective method of elution of pathogens. In this article, we report the benefits of incorporating megasonic sonication into the current methodologies of Giardia duodenalis elution from an internationally approved filtration and elution system used within the water industry, the Filta-Max®. Megasonic energy assisted elution has many benefits over current methods since a smaller final volume of eluent allows removal of time-consuming centrifugation steps and reduces manual involvement resulting in a potentially more consistent and more cost-effective method. We also show that megasonic sonication of G. duodenalis cysts provides the option of a less damaging elution method compared to the standard Filta-Max® operation, although the elution from filter matrices is not currently fully optimised. A notable decrease in recovery of damaged cysts was observed in megasonic processed samples, potentially increasing the abilities of further genetic identification options upon isolation of the parasite from a filter sample. This work paves the way for the development of a fully automated and more cost-effective elution method of Giardia from water samples.

Numerical study of the faithful replication of micro/nanostructures on curved surfaces by the electrohydrodynamic instability process.

This paper reports the numerical study of the one-step faithful replication of micro/nano-scale structures on a fiber surface by using the electrohydrodynamic instability patterning (EHDIP) process. By employing a rigorous numerical analysis method, conditions are revealed under which the faithful replication of a pattern can be achieved from a curved master electrode. It is found that the radius of curvature of the fiber plays an important role in determining the final morphology of the pattern when the destabilizing electric field is dominant in both the flat and patterned template cases. In general, stronger electric fields and larger radii of curvature of the substrate are favorable for the faithful replication of the pattern. In addition, theoretical analysis shows that higher aspect ratio of micro/nanostructures can be obtained on curved surfaces by using a master with a much lower aspect ratio. The results demonstrated in this study aims to provide guidelines for the faithful fabrication of micro/nanostructures on curved surfaces by the EHDIP process.

Autonomous capillary microfluidic system with embedded optics for improved troponin I cardiac biomarker detection.

Cardiovascular diseases are the most prevalent medical conditions affecting the modern world, reducing the quality of life for those affected and causing an ever increasing burden on clinical resources. Cardiac biomarkers are crucial in the diagnosis and management of patient outcomes. In that respect, such proteins are desirable to be measured at the point of care, overcoming the shortcomings of current instrumentation. We present a CO2 laser engraving technique for the rapid prototyping of a polymeric autonomous capillary system with embedded on-chip planar lenses and biosensing elements, the first step towards a fully miniaturised and integrated cardiac biosensing platform. The system has been applied to the detection of cardiac Troponin I, the gold standard biomarker for the diagnosis of acute myocardial infarction. The devised lab-on-a-chip device was demonstrated to have 24 pg/ml limit of detection, which is well within the minimum threshold for clinically applicable concentrations. Assays were completed within approximately 7-9 min. Initial results suggest that, given the portability, low power consumption and high sensitivity of the device, this technology could be developed further into point of care instrumentation useful in the diagnosis of various forms of cardiovascular diseases.

Validation of a fully integrated platform and disposable microfluidic chips enabling parallel purification of genome segments for assembly.

Recent progress in the field of genetic engineering has opened up the door to novel synthetic biology applications. Microfluidic technology has been emphasized as a key technology to support the development of these applications. While several important synthetic biology protocols have been developed in microfluidic format, no study has yet demonstrated on-chip error control. In synthetic biology protocols, the purification phase is a critical error control process which enhances the reliability of the genome segment assembly by removing undesired oligos. In this context, we report the design and characterization of a fully integrated platform, demonstrating the purification of up to 4 genome segments in parallel, prior to their off-chip assembly. The key innovation of this platform is the decoupling control strategy which eliminates the need to integrate expensive components onto the microfluidic device, enabling lower cost, disposability and rapid operation. Unlike most microfluidic chips where fluid connector plugs are needed to connect external pumps, this approach is plug-less and the chips are simply connected to the control breadboard by clamping. Furthermore the passive chip is isolated from the active control layer thereby eliminating the risk of sample-to-sample contamination in the reusable parts. As a validation of this fully-integrated system, the parallel on-chip purification of genome segments was demonstrated with ratio of correct phenotypes after final assembly up to 20% superior to the bench controls, proving thereby the suitability of the platform for synthetic biology applications.

Nonlinear hydrodynamic effects induced by Rayleigh surface acoustic wave in sessile droplets.

We report an experimental and numerical characterization of three-dimensional acoustic streaming behavior in small droplets of volumes (1-30 µl) induced by surface acoustic wave (SAW). We provide a quantitative evidence of the existence of strong nonlinear nature of the flow inertia in this SAW-driven flow over a range of the newly defined acoustic parameter F{NA}=Fλ/(σ/R_{d})≥0.01, which is a measure of the strength of the acoustic force to surface tension, where F is the acoustic body force, λ is the SAW wavelength, σ is the surface tension, and R{d} is the droplet radius. In contrast to the widely used Stokes model of acoustic streaming, which generally ignores such a nonlinearity, we identify that the full Navier-Stokes equation must be applied to avoid errors up to 93% between the computed streaming velocities and those from experiments as in the nonlinear case. We suggest that the Stokes model is valid only for very small acoustic power of ≤1 µW (F{NA}<0.002). Furthermore, we demonstrate that the increase of F{NA} above 0.45 induces not only internal streaming, but also the deformation of droplets.

Current and emerging techniques of fetal cell separation from maternal blood.

Intense research has been carried out in recent years into methods that aim to harvest fetal genetic material from maternal blood as substitutes to amniocentesis and chorionic villus sampling. Just over 30 years have past since the first fetal cells were separated from maternal blood using flow cytometry highlighting the prospect of non-invasive prenatal diagnosis of fetal abnormalities. The aim of this review paper is to describe the most commonly used cell separation methods with emphasis on the isolation of fetal cells from maternal blood. The most significant breakthroughs and advances in fetal cell separation are reviewed and critically analyzed. Although much has been accomplished using well established techniques, a rapid and inexpensive method to separate fetal cells with great accuracy, sensitivity and efficiency to maximize cell yield is still required. In the past decade MEMS (Micro Electro Mechanical Systems) technologies have enabled the miniaturization of many biological and medical laboratory processes. Lab-on-chip systems have been developed and encompass many modules capable of processing different biological samples. Such chips contain various integrated components such as separation channels, micropumps, mixers, reaction and detection chambers. This article will also explore new emerging MEMS based separation strategies, which hope to overcome the current limitations in fetal cell separation.

Recent advances in microparticle continuous separation.

Recent advances in microparticle separation in continuous flow are presented. It is intended for scientists in the field of separation science in biology, chemistry and microsystems engineering. Recent techniques of micron-sized particle separation within microsystems are described with emphasis on five different categories: optical, magnetic, fluidic-only, electrical and minor separation methods. Examples from the growing literature are explained with insights on separation efficiency and microengineering challenges. Current applications of the techniques are discussed.