Alexander Iles

Diamagnetic repulsion—A versatile tool for label-free particle handling in microfluidic devices

We report the exploration of diamagnetic repulsion forces for the selective manipulation of microparticles inside microfluidic devices. Diamagnetic materials such as polymers are repelled from magnetic fields, an effect greatly enhanced by suspending a diamagnetic object in a paramagnetic Mn(2+) solution. The versatility of diamagnetic repulsion is demonstrated for the trapping, focussing and deflection of polystyrene particles for three example applications. Firstly, magnet pairs with unlike poles facing each other were arranged along a microcapillary to trap plugs of differently functionalised particles for a simultaneous surface-based assay in which biotin was selectively bound to a plug of streptavidin coated particles utilising only 22nL of reagent. Secondly, by slightly modifying the magnetic field design, the rapid focussing of particles into a narrow central stream at a flow rate of 650microms(-1) was accomplished for particle pre-concentration. In a third application, 5 and 10microm polystyrene particles were separated from each other in continuous flow by passing the particle mixture through a microfluidic chamber with a perpendicular magnetic field, a method termed diamagnetophoresis. The separation was investigated between flow rates of 20-100microL h(-1), with full resolution of the particle populations being achieved at 20microL h(-1). These experiments show the potential of diamagnetic repulsion for simple, label-free manipulation of particles and other diamagnetic objects such as cells for a range of bioanalytical techniques.

Rapid on-chip multi-step (bio)chemical procedures in continuous flow : manoeuvring particles through co-laminar reagent streams

We introduce a novel and extremely versatile microfluidic platform in which tedious multi-step biochemical processes can be performed in continuous flow within a fraction of the time required for conventional methods.

Rapid, multistep on-chip DNA hybridisation in continuous flow on magnetic particles.

DNA hybridisation is an important tool for bioanalytical research and clinical diagnostics; conventional methods, however, require long incubation times and numerous washing steps, rendering the procedure time consuming and labour intensive. In this paper, we report on a rapid method for DNA hybridisation and isolation within a microfluidic device, where all reaction and washing steps are performed in continuous flow in an automated fashion within less than two minutes. Magnetic particles were used as a solid support and manipulated through laminar flow streams containing reagents and buffers by means of an external magnet. Thus, hybridisation, washing, intercalation, fluorescence detection and isolation were performed in continuous flow on the surface of the particles. Initially, the sensitivity of the system was investigated for a one-step DNA hybridisation of Alexa Fluor 555 labelled target DNA to a capture probe immobilised on the particle surface. Hybridisation and washing steps were performed in half a minute and target DNA was readily detected down to 20 nmol L(-1). Then a two-step assay, label-free DNA hybridisation followed by intercalation with PicoGreen was performed. All reaction and washing steps were carried out in continuous flow with a total assay time of about 1 min. This is a significant reduction in procedural time compared to conventional methods and opens the door for developing fully automated continuous flow integrated DNA analysis platforms.

On-chip diamagnetic repulsion in continuous flow

Abstract We explore the potential of a microfluidic continuous flow particle separation system based on the repulsion of diamagnetic materials from a high magnetic field. Diamagnetic polystyrene particles in paramagnetic manganese (II) chloride solution were pumped into a microfluidic chamber and their deflection behaviour in a high magnetic field applied by a superconducting magnet was investigated. Two particle sizes (5 and 10 μm) were examined in two concentrations of MnCl2 (6 and 10%). The larger particles were repelled to a greater extent than the smaller ones, and the effect was greatly enhanced when the particles were suspended in a higher concentration of MnCl2. These findings indicate that the system could be viable for the separation of materials of differing size and/or diamagnetic susceptibility, and as such could be suitable for the separation and sorting of small biological species for subsequent studies.

Sorting and Manipulation of Magnetic Droplets in Continuous Flow

We report the rapid on‐chip generation and subsequent manipulation of magnetic droplets in continuous flow. Magnetic droplets were formed using aqueous‐based ferrofluid as the dispersed phase and fluorocarbon oil as the continuous phase. Droplet manipulation was demonstrated with simple permanent magnets using two microfluidic platforms: (i) flow focusing droplet generation followed by their splitting into daughter droplets containing different amounts of magnetic nanoparticles, and (ii) droplet generation at a T‐junction and their downstream deflection across a chamber for sorting based on the applied magnetic field and magnetite loading of the droplet. Both systems show great potential for performing a wide range of high throughput continuous flow processes including sample dilution, cell sorting and screening, and microparticle fabrication.

Microfluidic Technique for the Simultaneous Quantification of Emulsion Instabilities and Lipid Digestion Kinetics

Quantifying the impact of environmental physicochemical changes on the microstructure of lipid delivery systems is challenging. Therefore, we have developed a methodology to quantify the coalescence of oil-in-water emulsion droplets during lipid digestion in situ on a single droplet level. This technique involves a custom-made glass microfluidic platform, in which oil droplets can be trapped as single droplets, or several droplets per trap. The physicochemical environment can be controlled, and droplet digestion, as well as coalescence, can be visualized. We show that the exchange of the physicochemical conditions in the entire reaction chamber can be reached in under 30 s. Microparticle image velocimetry allowed mapping of the flow profile and demonstrated the tuneability of the shear profile in the device. The extraction of quantitative information regarding the physical characteristics of the droplets during digestion was performed using an automated image analysis throughout the digestion process. Therefore, we were able to show that oil-in-water emulsions stabilized by proteins coalesced under human gastric conditions. This coalescence delayed the overall lipid digestion kinetics. The droplets that coalesced during digestion were hydrolyzed 1.4 times slower than individually trapped droplets. Thus, the microstructural evolution of lipid delivery systems is a crucial factor in lipid digestion kinetics. This novel technique allows the simultaneous quantification of the impact that the physicochemical environment has on both the lipid droplet microstructure and the lipid release patterns.

A Microfluidic Device for Rapid Screening of E. coli O157:H7 Based on IFAST and ATP Bioluminescence Assay for Water Analysis

We present a simple microfluidic system for rapid screening of Escherichia coli (E. coli) O157:H7 employing the specificity of immunomagnetic separation (IMS) via immiscible filtration assisted by surface tension (IFAST), and the sensitivity of the subsequent adenosine triphosphate (ATP) assay by the bioluminescence luciferin/luciferase reaction. The developed device was capable of detecting E. coli O157:H7 from just 6 colony forming units (CFU) in 1 mL spiked buffer within 20 min. When tested with wastewater discharged effluent samples, without pre-concentration, the device demonstrated the ability to detect 104  CFU per mL seeded; suggesting great potential for point-of-need microbiological water quality monitoring.

Bonding of Soda-Lime Glass Microchips at Low Temperature

The bonding of glass wafers is an extremely important aspect of microfluidic chip fabrication. Conventionally this is performed using high temperatures over long periods of time. This is time consuming and energy intensive and can have detrimental effects on the quality of the completed device. In this work we have adapted the low temperature HF bonding method that has been used for quartz so that it can be applied to soda lime glass, the most commonly used chip fabrication material. A range of devices were fabricated using this method and the quality of the bond interface was examined using SEM-EDX measurements.