29 resultados para lab-on-a-chip

em Aston University Research Archive


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Immunoprecipitation (IP) is one of the most widely used and selective techniques for protein purification. Here, a miniaturised, polymer-supported immunoprecipitation (µIP) method for the on-chip purification of proteins from complex mixtures is described. A 4 µl PDMS column functionalised with covalently bound antibodies was created and all critical aspects of the µIP protocol (antibody immobilisation, blocking of potential non-specific adsorption sites, sample incubation and washing conditions) were assessed and optimised. The optimised µIP method was used to obtain purified fractions of affinity-tagged protein from a bacterial lysate.

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Nanoparticles offer an ideal platform for the delivery of small molecule drugs, subunit vaccines and genetic constructs. Besides the necessity of a homogenous size distribution, defined loading efficiencies and reasonable production and development costs, one of the major bottlenecks in translating nanoparticles into clinical application is the need for rapid, robust and reproducible development techniques. Within this thesis, microfluidic methods were investigated for the manufacturing, drug or protein loading and purification of pharmaceutically relevant nanoparticles. Initially, methods to prepare small liposomes were evaluated and compared to a microfluidics-directed nanoprecipitation method. To support the implementation of statistical process control, design of experiment models aided the process robustness and validation for the methods investigated and gave an initial overview of the size ranges obtainable in each method whilst evaluating advantages and disadvantages of each method. The lab-on-a-chip system resulted in a high-throughput vesicle manufacturing, enabling a rapid process and a high degree of process control. To further investigate this method, cationic low transition temperature lipids, cationic bola-amphiphiles with delocalized charge centers, neutral lipids and polymers were used in the microfluidics-directed nanoprecipitation method to formulate vesicles. Whereas the total flow rate (TFR) and the ratio of solvent to aqueous stream (flow rate ratio, FRR) was shown to be influential for controlling the vesicle size in high transition temperature lipids, the factor FRR was found the most influential factor controlling the size of vesicles consisting of low transition temperature lipids and polymer-based nanoparticles. The biological activity of the resulting constructs was confirmed by an invitro transfection of pDNA constructs using cationic nanoprecipitated vesicles. Design of experiments and multivariate data analysis revealed the mathematical relationship and significance of the factors TFR and FRR in the microfluidics process to the liposome size, polydispersity and transfection efficiency. Multivariate tools were used to cluster and predict specific in-vivo immune responses dependent on key liposome adjuvant characteristics upon delivery a tuberculosis antigen in a vaccine candidate. The addition of a low solubility model drug (propofol) in the nanoprecipitation method resulted in a significantly higher solubilisation of the drug within the liposomal bilayer, compared to the control method. The microfluidics method underwent scale-up work by increasing the channel diameter and parallelisation of the mixers in a planar way, resulting in an overall 40-fold increase in throughput. Furthermore, microfluidic tools were developed based on a microfluidics-directed tangential flow filtration, which allowed for a continuous manufacturing, purification and concentration of liposomal drug products.

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This thesis documents the design, manufacture and testing of a passive and non-invasive micro-scale planar particle-from-fluid filter for segregating cell types from a homogeneous suspension. The microfluidics system can be used to separate spermatogenic cells from testis biopsy samples, providing a mechanism for filtrate retrieval for assisted reproduction therapy. The system can also be used for point-of-service diagnostics applications for hospitals, lab-on-a-chip pre-processing and field applications such as clinical testing in the third world. Various design concepts are developed and manufactured, and are assessed based on etched structure morphology, robustness to variations in the manufacturing process, and design impacts on fluid flow and particle separation characteristics. Segregation was measured using image processing algorithms that demonstrate efficiency is more than 55% for 1 µl volumes at populations exceeding 1 x 107. the technique supports a significant reduction in time over conventional processing, in the separation and identification of particle groups, offering a potential reduction in the associated cost of the targeted procedure. The thesis has developed a model of quasi-steady wetting flow within the micro channel and identifies the forces across the system during post-wetting equalisation. The model and its underlying assumptions are validated empirically in microfabricated test structures through a novel Micro-Particle Image Velocimetry technique. The prototype devices do not require ancillary equipment nor additional filtration media, and therefore offer fewer opportunities for sample contamination over conventional processing methods. The devices are disposable with minimal reagent volumes and process waste. Optimal processing parameters and production methods are identified with any improvements that could be made to enhance their performance in a number of identified potential applications.

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Ultrasonics offers the possibility of developing sophisticated fluid manipulation tools in lab-on-a-chip technologies. Here we demonstrate the ability to shape ultrasonic fields by using phononic lattices, patterned on a disposable chip, to carry out the complex sequence of fluidic manipulations required to detect the rodent malaria parasite Plasmodium berghei in blood. To illustrate the different tools that are available to us, we used acoustic fields to produce the required rotational vortices that mechanically lyse both the red blood cells and the parasitic cells present in a drop of blood. This procedure was followed by the amplification of parasitic genomic sequences using different acoustic fields and frequencies to heat the sample and perform a real-time PCR amplification. The system does not require the use of lytic reagents nor enrichment steps, making it suitable for further integration into lab-on-a-chip point-of-care devices. This acoustic sample preparation and PCR enables us to detect ca. 30 parasites in a microliter-sized blood sample, which is the same order of magnitude in sensitivity as lab-based PCR tests. Unlike other lab-on-a-chip methods, where the sample moves through channels, here we use our ability to shape the acoustic fields in a frequency-dependent manner to provide different analytical functions. The methods also provide a clear route toward the integration of PCR to detect pathogens in a single handheld system.

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Besides their well-described use as delivery systems for water-soluble drugs, liposomes have the ability to act as a solubilizing agent for drugs with low aqueous solubility. However, a key limitation in exploiting liposome technology is the availability of scalable, low-cost production methods for the preparation of liposomes. Here we describe a new method, using microfluidics, to prepare liposomal solubilising systems which can incorporate low solubility drugs (in this case propofol). The setup, based on a chaotic advection micromixer, showed high drug loading (41 mol%) of propofol as well as the ability to manufacture vesicles with at prescribed sizes (between 50 and 450 nm) in a high-throughput setting. Our results demonstrate the ability of merging liposome manufacturing and drug encapsulation in a single process step, leading to an overall reduced process time. These studies emphasise the flexibility and ease of applying lab-on-a-chip microfluidics for the solubilisation of poorly water-soluble drugs.

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In this paper, we study generation of Bessel beams from semiconductor lasers with high beam propagation parameter M2 and their utilization for optical trapping and manipulation of microscopic particles including living cells. The demonstrated optical tweezing with diodegenerated Bessel beams paves the way to replace their vibronic-generated counterparts for a range of applications towards novel lab-on-a-chip configurations.

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We report on recent progress in the generation of non-diffracting (Bessel) beams from semiconductor light sources including both edge-emitting and surface-emitting semiconductor lasers as well as light-emitting diodes (LEDs). Bessel beams at the power level of Watts with central lobe diameters of a few to tens of micrometers were achieved from compact and highly efficient lasers. The practicality of reducing the central lobe size of the Bessel beam generated with high-power broad-stripe semiconductor lasers and LEDs to a level unachievable by means of traditional focusing has been demonstrated. We also discuss an approach to exceed the limit of power density for the focusing of radiation with high beam propagation parameter M2. Finally, we consider the potential of the semiconductor lasers for applications in optical trapping/tweezing and the perspectives to replace their gas and solid-state laser counterparts for a range of implementations in optical manipulation towards lab-on-chip configurations. © 2014 Elsevier Ltd.

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Many applications of high-power laser diodes demand tight focusing. This is often not possible due to the multimode nature of semiconductor laser radiation possessing beam propagation parameter M2 values in double-digits. We propose a method of 'interference' superfocusing of high-M2 diode laser beams with a technique developed for the generation of Bessel beams based on the employment of an axicon fabricated on the tip of a 100 μm diameter optical fiber with highprecision direct laser writing. Using axicons with apex angle 140º and rounded tip area as small as 10 μm diameter, we demonstrate 2-4 μm diameter focused laser 'needle' beams with approximately 20 μm propagation length generated from multimode diode laser with beam propagation parameter M2=18 and emission wavelength of 960 nm. This is a few-fold reduction compared to the minimal focal spot size of 11 μm that could be achieved if focused by an 'ideal' lens of unity numerical aperture. The same technique using a 160º axicon allowed us to demonstrate few-μm-wide laser 'needle' beams with nearly 100 μm propagation length with which to demonstrate optical trapping of 5-6 μm rat blood red cells in a water-heparin solution. Our results indicate the good potential of superfocused diode laser beams for applications relating to optical trapping and manipulation of microscopic objects including living biological objects with aspirations towards subsequent novel lab-on-chip configurations.

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An important parameter in integrated optical device is the propagation loss of the waveguide. Its characterization gives the information of the fabrication quality as well as the information of other passive devices on the chip as it is the basic building block of the passive devices. Although, over the last three decades many methods have been developed, there is not a single standard present yet. This paper presents a comparative analysis of the methods existing from the past as well as methods developed very recently in order to provide a complete picture of the pros and cons of different types of methods and from this comparison the best method is suggested according to the authors opinion. To support the claim, apart from the analytical comparison, this paper also presents a comparison performed with the experimental results between the suggested best method which is recently proposed by Massachusetts Institute of Technology (MIT) researchers based on undercoupled all-pass microring structure and the popular cut-back method.

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As mobile technologies continue to penetrate increasingly diverse domains of use, we accordingly need to understand the feasibility of different interaction technologies across such varied domains. This case study describes an investigation into whether speechbased input is a feasible interaction option for use in a complex, and arguably extreme, environment of use – that is, lobster fishing vessels. We reflect on our approaches to bringing the “high seas” into lab environments for this purpose, comparing the results obtained via our lab and our field studies. Our hope is that the work presented here will go some way to enhancing the literature in terms of approaches to bringing complex real-world contexts into lab environments for the purpose of evaluating the feasibility of specific interaction technologies.

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As mobile technologies continue to penetrate increasingly diverse domains of use, we accordingly need to understand the feasibility of different interaction technologies across such varied domains. This case study describes an investigation into whether speechbased input is a feasible interaction option for use in a complex, and arguably extreme, environment of use – that is, lobster fishing vessels. We reflect on our approaches to bringing the “high seas” into lab environments for this purpose, comparing the results obtained via our lab and our field studies. Our hope is that the work presented here will go some way to enhancing the literature in terms of approaches to bringing complex real-world contexts into lab environments for the purpose of evaluating the feasibility of specific interaction technologies.

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DUE TO COPYRIGHT RESTRICTIONS ONLY AVAILABLE FOR CONSULTATION AT ASTON UNIVERSITY LIBRARY AND INFORMATION SERVICES WITH PRIOR ARRANGEMENT

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This paper details methodologies that have been explored for the fast proofing of on-chip architectures for Circular Dichroism techniques. Flow-cell devices fabricated from UV transparent Quartz are used for these experiments. The complexity of flow-cell production typically results in lead times of six months from order to delivery. Only at that point can the on-chip architecture be tested empirically and any required modifications determined ready for the next six month iteration phase. By using the proposed 3D printing and PDMS moulding techniques for fast proofing on-chip architectures the optimum design can be determined within a matter of hours prior to commitment to quartz chip production.

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This paper focuses on minimizing printed circuit board (PCB) assembly time for a chipshootermachine, which has a movable feeder carrier holding components, a movable X–Y table carrying a PCB, and a rotary turret with multiple assembly heads. The assembly time of the machine depends on two inter-related optimization problems: the component sequencing problem and the feeder arrangement problem. Nevertheless, they were often regarded as two individual problems and solved separately. This paper proposes two complete mathematical models for the integrated problem of the machine. The models are verified by two commercial packages. Finally, a hybrid genetic algorithm previously developed by the authors is presented to solve the model. The algorithm not only generates the optimal solutions quickly for small-sized problems, but also outperforms the genetic algorithms developed by other researchers in terms of total assembly time.

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A chip shooter machine for electronic component assembly has a movable feeder carrier, a movable X–Y table carrying a printed circuit board (PCB), and a rotary turret with multiple assembly heads. This paper presents a hybrid genetic algorithm (HGA) to optimize the sequence of component placements and the arrangement of component types to feeders simultaneously for a chip shooter machine, that is, the component scheduling problem. The objective of the problem is to minimize the total assembly time. The GA developed in the paper hybridizes different search heuristics including the nearest-neighbor heuristic, the 2-opt heuristic, and an iterated swap procedure, which is a new improved heuristic. Compared with the results obtained by other researchers, the performance of the HGA is superior in terms of the assembly time. Scope and purpose When assembling the surface mount components on a PCB, it is necessary to obtain the optimal sequence of component placements and the best arrangement of component types to feeders simultaneously in order to minimize the total assembly time. Since it is very difficult to obtain the optimality, a GA hybridized with several search heuristics is developed. The type of machines being studied is the chip shooter machine. This paper compares the algorithm with a simple GA. It shows that the performance of the algorithm is superior to that of the simple GA in terms of the total assembly time.