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This book describes the continuing development of inexpensive, portable flow cytometers through incorporation of microfluidic technologies and small optical components. The underlying microfluidic theories essential for microflow cytometry isdiscussed in detail, as well as advances that are representative of the current state-of-the-art. Design and fabrication strategies for these innovative component technologies will be subsequently presented by numerous research groups leading the field. Integration of the components into functional prototype devices for analysis and manipulation of particles and cells are reviewed. Multiple currently available commercial systems are examined to highlight both strengths and areas for improvement.
This book describes novel microtechnologies and integration strategies for developing a new class of assay systems to retrieve desired health information from patients in real-time. The selection and integration of sensor components and operational parameters for developing point-of-care (POC) are also described in detail. The basics that govern the microfluidic regimen and the techniques and methods currently employed for fabricating microfluidic systems and integrating biosensors are thoroughly covered. This book also describes the application of microfluidics in the field of cell and molecular biology, single cell biology, disease diagnostics, as well as the commercially available systems that have been either introduced or have the potential of being used in research and development. This is an ideal book for aiding biologists in understanding the fundamentals and applications of microfluidics. This book also: Describes the preparatory methods for developing 3-dimensional microfluidic structures and their use for Lab-on-a-Chip design Explains the significance of miniaturization and integration of sensing components to develop wearable sensors for point-of-care (POC) Demonstrates the application of microfluidics to life sciences and analytical chemistry, including disease diagnostics and separations Motivates new ideas related to novel platforms, valving technology, miniaturized transduction methods, and device integration to develop next generation sequencing Discusses future prospects and challenges of the field of microfluidics in the areas of life sciences in general and diagnostics in particular
My thesis research has focused on means of integrating optical systems into microfluidic chips, specifically for the creation of lab-on-a-chip flow cytometers. The benefits of microfluidics are perhaps most often applied to biological assays, which frequently employ optical readout of fluorescence or light scatter. By integrating the optical system onto the microfluidic chip, we can facilitate chip interfacing while ensuring optical alignment to a tiny sample. Integrated optical systems also offer the ability to collect light from a localized area, allowing for the collection of true angular light scatter (which carries much information about cells) and can furthermore significantly improve the signal to noise ratio (SNR) relative to simple fiber or waveguide based approaches to integrated light collection. This work explores both the unique challenges and advantages encountered when creating optical systems integrated with mold-replicated microfluidic devices. The first contribution presented is the demonstration of fluid-filled lenses integrated alongside microfluidic channels using a slab waveguiding structure. The use of fluid represents an important tradeoff between lens power and Fresnel reflections. The creation of a slab waveguiding structure is critically important to control light losses when utilizing lens systems for light collection. The second contribution in this work is the demonstration of a microfluidic chip emplying a number of lenses to perform both localized excitation of the samples as well as light collection from localized areas defined by a specific angular range. Sample coefficients of variation (CVs) ranged from 9-16% for a single bead population, far exceeding previously-published CVs of 25-35%. The last contribution is an atypical approach to optical systems based on the unique advantages offered by microfabricated architectures, namely small sizes and close proximities to the sample. Using only custom-shaped total internal reflection (TIR) based components and light blocking elements, we create a device that can achieve forward light scatter CVs of 8-28%. The device is able to clearly distinguish 5 @mu;m, 10 @mu;m, and 15 @mu;m beads based on forward and side scatter. The results from this vastly simplified optical system show great promise towards reaching the performance metrics of the commercial cytometer.
Microfluidics provides a unique and useful platform for performing analytical measurements. Micro-optics can be imbedded into a microfluidic device, and the analyte can be maneuvered hydrodynamically into the optical interrogation zone. In addition, since flow is laminar, the concentration distributions of the solutes can be explicitly calculated over the volume of the device. In this work, a microfluidic hydrodynamic focusing manifold was developed which focused a solution into the center of a square microfluidic channel. By adjusting the flow-rates of the focusing fluids, the solution-under-focusing was moved up and down relative to the top and bottom walls of the microchannel. In one embodiment, the hydrodynamic focusing manifold was integrated with imbedded optical fibers for use as a micro- flow cytometer. By hydrodynamically aligning the particle stream with the laser beam, absolute counting efficiency of 58±8% was demonstrated for fluorescent microparticles at a throughput of 5.5 [MICRO SIGN]L/min of bead solution. This device can be used to analyze a multiplexed bead-based assay for point-of-detection biosensing. In a second embodiment, the focusing manifold fed into a long microchannel where solute-mixing took place under pressure-driven laminar flow conditions. A numerical analysis program was developed which modeled the three-dimensional microfluidic channel with a two-dimensional mesh by using the moving mesh method. Using this program, concentration distributions were calculated at distances along the microchannel, and these were compared to an imaging experiment. The numerical analysis program overestimated the diffusion coefficients of fluorescein and Enhanced Green Fluorescent Protein by factors of 1.9 ± 0.4 and 1.4 ± 0.2, respectively. The source of this overestimation was hydrodynamic dispersion, the effect of which could not be properly treated using the moving mesh method due to the assumption transverse-uniform fluid-velocity over the cross-section of the microchannel. Advances in numerical analysis are needed for improved modeling and characterization of microfluidic devices, particularly those for which the two effects of hydrodynamic dispersion and molecular diffusion simultaneously influence the movements of the solutes. This dissertation informs the development of microfluidic analytical devices and the analysis of solute-transport within microfluidic channels.
The concept behind this book is to provide a detailed and practical overview of the development and use of immunoassays in many different areas. Immunoassays are analytical tests that utilise antibodies to measure the amount, activity or identity of an analyte. This book is designed to provide a critical and helpful insight into the subject and to give the user practical information that may be of assistance in assay format selection, antibody generation/selection and choice of appropriate detection strategies. It is comprised of 13 chapters written by highly experienced researchers in the fields of antibody-based research, immunoassay development, assay validation, diagnostics and microfluidics. Beginning with a comprehensive survey of antibodies, immunoassay formats and signalling systems, the book elucidates key topics related to the development of an ideal antibody-based sensor, focuses on the important topic of surface modification, explores key parameters in the immobilisation of antibodies onto solid surfaces, discusses the move to ‘lab-on-a-chip’-based devices and investigates the key parameters necessary for their development. Three of the chapters are dedicated to the areas of clinical diagnostics, infectious disease monitoring and food security, where immunoassay-based applications have become highly valuable tools. The future of immunoassays, including next-generation immunoassays, electrochemical-immunoassays and ‘lab-on-a-chip’-based systems, is also discussed. The book also covers the use of optical detection systems (with a focus on surface plasmon resonance) in immunoassays, provides a compilation of important, routinely used immunoassay protocols and addresses problems that may be encountered during assay development.
Comprehensive Biotechnology, Third Edition, Six Volume Set unifies, in a single source, a huge amount of information in this growing field. The book covers scientific fundamentals, along with engineering considerations and applications in industry, agriculture, medicine, the environment and socio-economics, including the related government regulatory overviews. This new edition builds on the solid basis provided by previous editions, incorporating all recent advances in the field since the second edition was published in 2011. Offers researchers a one-stop shop for information on the subject of biotechnology Provides in-depth treatment of relevant topics from recognized authorities, including the contributions of a Nobel laureate Presents the perspective of researchers in different fields, such as biochemistry, agriculture, engineering, biomedicine and environmental science
Surface plasmon resonance (SPR) plays a dominant role in real-time interaction sensing of biomolecular binding events, this book provides a total system description including optics, fluidics and sensor surfaces for a wide researcher audience.
This book describes the emerging point-of-care (POC) technologies that are paving the way to the next generation healthcare monitoring and management. It provides the readers with comprehensive, up-to-date information about the emerging technologies, such as smartphone-based mobile healthcare technologies, smart devices, commercial personalized POC technologies, paper-based immunoassays (IAs), lab-on-a-chip (LOC)-based IAs, and multiplex IAs. The book also provides guided insights into the POC diabetes management software and smart applications, and the statistical determination of various bioanalytical parameters. Additionally, the authors discuss the future trends in POC technologies and personalized and integrated healthcare solutions for chronic diseases, such as diabetes, stress, obesity, and cardiovascular disorders. Each POC technology is described comprehensively and analyzed critically with its characteristic features, bioanalytical principles, applications, advantages, limitations, and future trends. This book would be a very useful resource and teaching aid for professionals working in the field of POC technologies, in vitro diagnostics (IVD), mobile healthcare, Big Data, smart technology, software, smart applications, biomedical engineering, biosensors, personalized healthcare, and other disciplines.