37 research outputs found

    Microfluidic Technology and Application in Urinal Analysis

    Get PDF
    Microfluidic technology offers numerous advantages in minimizing and integrating the traditional assays. However, the lack of efficient control components of the microfluidic systems has been hindering the widely commercialization of the technology. The research work in this dissertation focused on the development of effective control components for microfluidic applications. A linear peristaltic pump was firstly designed, fabricated, and tested for conventional microfluidics by synchronously compressing the microfluidic channel with a miniature cam-follower system in Chapter 2. The miniature cam-follower system and microfluidic chip was prototyped using three-dimensional (3D) printing technology and soft lithography technology. Results from experimental test showed that the pump is self-priming and tolerant of bubbles. The pumping flowrate and back pressure could be controlled by changing the driving speed of the motor. Then a novel pinch-type valving system that can be used to realize both normally closed and normally open valves for centrifugal microfluidics was demonstrated in Chapter 3. A sliding wedge was actuated by centrifugal force to drive the valves. Experimental test and theoretical predication showed that the burst frequency of the valves could be tuned by changing the physical parameters of the valving system. In Chapter 4, the pinch type valving system was then further improved for better integration of multiple valves in limited space to realize sequential control of microfluidics. A valve chip with grooves on the surface was used to drive multiple valves. A flow switch which is capable of working at low rotation frequency and constant rotation direction is realized. Finally, the microfluidic platform was utilized for automatic urinalysis for the application at point of care (POC) to eliminate the difficulties in control of sample distribution and read-out time in manually conducted colorimetric urinalysis. 3D printed prototype of the microfluidic chip was used to test the proposed system. Commercial urinalysis strips was integrated with the microfluidic system for detecting glucose, specific gravity, PH, and protein from simulated urine sample. The color change of the pads was recorded using smartphone camera and analyzed to quantify the interested parameters

    Organ-on-a-Disc: A Scalable Platform Technology for the Generation and Cultivation of Microphysiological Tissues

    Get PDF
    Organ-on-Chip (OoC) systems culture human tissues in a controllable environment under microfluidic perfusion and enable a precise recapitulation of human physiology. Although recent studies demonstrate the potential of OoCs as alternative to traditional cell assays and animal models in drug development as well as personalized medicine, unmet challenges in device fabrication, parallelization and operation hinder their widespread application. In order to overcome these obstacles, this thesis focuses on the development of the Organ-on-a-Disc technology for the scalable generation and cultivation of microphysiological tissues. Organ-Discs are fabricated using precise, rapid and scalable microfabrication techniques. They enable the pump- and tubing-free perfusion as well as the parallelized generation and culture of tailorable and functional microtissues using rotation-based operations. The Organ-Disc setup is suitable for versatile tissue readouts, treatments and even whole blood perfusion with minimal handling and equipment requirements. Overall, the Organ-Disc creates a scalable and userfriendly platform technology for microphysiological tissue models and paves the way for their transition towards high-throughput systems.:Abbreviations Symbols 1 Introduction 2 Background 2.1 Fluid Dynamics 2.1.1 Flow Equations 2.1.2 Hydraulic Resistance 2.1.3 Wall Shear Stress 2.1.4 Centrifugal Microfluidics 2.2 Microfluidic Chip Fabrication 2.2.1 Chip Materials 2.2.2 Microstructuring 2.2.3 Bonding 3 State of the Art 3.1 Cell Culture Systems 3.2 3D Tissue Generation in Microfluidic Systems 3.3 Organ-on-Chip 3.4 Scale-up of Organ-on-Chip Systems 3.4.1 Scalable Fabrication Technologies 3.4.2 Parallelization Approaches 3.4.3 Integrated Fluid Actuation 3.5 Centrifugal Microfluidics 4 Objectives 5 Materials and Methods 5.1 Organ-Disc Fabrication 5.1.1 Materials 5.1.2 2D Structuring 5.1.3 Hot Embossing Stamp Fabrication TPE Hot Embossing 5.1.4 Bonding Solvent Vapor Bonding Thermal Fusion Bonding TPE Bonding 5.1.5 Characterization Methods Structure Sizes Bonding Strength Optical Properties 5.2 Organ-Disc Spinner 5.2.1 Centrifugal Loading Setup 5.2.2 Centrifugal Perfusion Setup 5.2.3 Peristaltic Pumping Setup 5.3 Organ-Disc Perfusion 5.3.1 Centrifugal Perfusion 5.3.2 Peristaltic Perfusion 5.4 Preparatory Cell Culture 5.5 Organ-Disc Cell Loading 5.5.1 Centrifugal Cell Loading 5.5.2 Endothelial-lining 5.6 Organ-Disc Cell Culture 5.6.1 Staining and Imaging Live Cell Labeling Live/Dead Staining CD106 Staining CD41 Staining Fixation, Permeabilization and Blocking Actin/Nuclei Staining CD31/Nuclei Staining 5.6.2 Media Analysis 5.6.3 Endothelial Cell Activation 5.6.4 Whole Blood Perfusion 5.7 Data Presentation and Statistics 6 Concept and Design 6.1 Organ-Disc Technology 6.2 Organ-Disc Design 6.3 Centrifugal Cell Loading 6.4 Endothelial Cell Lining 6.5 Centrifugal Perfusion 6.6 Peristaltic Perfusion 7 Building Blocks 7.1 Microfabrication Technology 7.1.1 Structuring 2D Structuring Hot Embossing 7.1.2 Bonding Solvent Vapor Bonding Thermal Fusion Bonding TPE Bonding 7.2 Organ-Disc Spinner 8 Perfusion 8.1 Centrifugal Pumping 8.2 Peristaltic Pumping 9 Tissue Generation and Culture 9.1 3D Tissue Generation 9.2 Stratified Tissue Construction 9.3 Generation of Endothelial-lined Channels 9.4 Perfusion of Endothelial-lined Channels 9.4.1 Media Monitoring Evaporation Cell Metabolism 9.4.2 Inflammatory Cell Stimulation 9.4.3 Whole Blood Perfusion 10 Discussion 10.1 Organ-Disc Technology 10.2 Scalable, Precise and Robust Organ-Disc Fabrication 10.2.1 Fabrication of Thermoplastic Organ-Discs 10.2.2 Fabrication of TPE Modules 10.2.3 Integration of TPE Modules to Organ-Discs 10.3 Tunable, Pump- and Tubing-free Perfusion 10.4 On-Disc Tissue Culture 10.4.1 3D Tissues 10.4.2 Blood Vessel-like Structures 10.4.3 Tissue Characterization and Treatment 10.5 On-Disc Blood Perfusion 11 Summary and Conclusion 12 References 13 AppendixIn Organ-on-Chip (OoC)-Systemen werden menschliche Gewebe mittels mikrofluidischer Versorgung in einer kontrollierten Umgebung kultiviert und so die Physiologie des Menschen nachgebildet. Obwohl aktuelle Studien zeigen, dass dieser Ansatz Alternativen zu herkömmlichen Zellbasierten Tests und Tiermodellen in der Arzneimittelentwicklung und der personalisierten Medizin bietet, stehen einer breiteren Anwendung Hürden im Bereich der Herstellung, Parallelisierung und Handhabung im Weg. Deshalb ist das Ziel dieser Arbeit die Entwicklung der Organ-on-a-Disc-Technologie, die eine skalierbare Erzeugung und Kultur von mikrophysiologischen Geweben ermöglicht. Für die Herstellung von der Organ-Disc kommen präzise, schnelle und skalierbare Mikrofabrikationsmethoden zum Einsatz. Die Organ-Disc schafft die Basis für die parallelisierte Erzeugung und Kultur von maßgeschneiderten und funktionellen Mikrogeweben, sowie deren Versorgung durch rotationsbasierte Prozesse und ohne zur Hilfenahme von Pumpen oder Schläuchen. Die Organ-Disc eignet sich für unterschiedliche Charakterisierungsmethoden sowie der Gewebestimulation und sogar der Vollblutperfusion mit minimalem Aufwand und Equipment. Insgesamt stellt die Organ-Disc eine skalierbare und benutzerfreundliche Plattformtechnologie für mikrophysiologische Modelle dar und bereitet den Weg für Hochdurchsatzanwendungen.:Abbreviations Symbols 1 Introduction 2 Background 2.1 Fluid Dynamics 2.1.1 Flow Equations 2.1.2 Hydraulic Resistance 2.1.3 Wall Shear Stress 2.1.4 Centrifugal Microfluidics 2.2 Microfluidic Chip Fabrication 2.2.1 Chip Materials 2.2.2 Microstructuring 2.2.3 Bonding 3 State of the Art 3.1 Cell Culture Systems 3.2 3D Tissue Generation in Microfluidic Systems 3.3 Organ-on-Chip 3.4 Scale-up of Organ-on-Chip Systems 3.4.1 Scalable Fabrication Technologies 3.4.2 Parallelization Approaches 3.4.3 Integrated Fluid Actuation 3.5 Centrifugal Microfluidics 4 Objectives 5 Materials and Methods 5.1 Organ-Disc Fabrication 5.1.1 Materials 5.1.2 2D Structuring 5.1.3 Hot Embossing Stamp Fabrication TPE Hot Embossing 5.1.4 Bonding Solvent Vapor Bonding Thermal Fusion Bonding TPE Bonding 5.1.5 Characterization Methods Structure Sizes Bonding Strength Optical Properties 5.2 Organ-Disc Spinner 5.2.1 Centrifugal Loading Setup 5.2.2 Centrifugal Perfusion Setup 5.2.3 Peristaltic Pumping Setup 5.3 Organ-Disc Perfusion 5.3.1 Centrifugal Perfusion 5.3.2 Peristaltic Perfusion 5.4 Preparatory Cell Culture 5.5 Organ-Disc Cell Loading 5.5.1 Centrifugal Cell Loading 5.5.2 Endothelial-lining 5.6 Organ-Disc Cell Culture 5.6.1 Staining and Imaging Live Cell Labeling Live/Dead Staining CD106 Staining CD41 Staining Fixation, Permeabilization and Blocking Actin/Nuclei Staining CD31/Nuclei Staining 5.6.2 Media Analysis 5.6.3 Endothelial Cell Activation 5.6.4 Whole Blood Perfusion 5.7 Data Presentation and Statistics 6 Concept and Design 6.1 Organ-Disc Technology 6.2 Organ-Disc Design 6.3 Centrifugal Cell Loading 6.4 Endothelial Cell Lining 6.5 Centrifugal Perfusion 6.6 Peristaltic Perfusion 7 Building Blocks 7.1 Microfabrication Technology 7.1.1 Structuring 2D Structuring Hot Embossing 7.1.2 Bonding Solvent Vapor Bonding Thermal Fusion Bonding TPE Bonding 7.2 Organ-Disc Spinner 8 Perfusion 8.1 Centrifugal Pumping 8.2 Peristaltic Pumping 9 Tissue Generation and Culture 9.1 3D Tissue Generation 9.2 Stratified Tissue Construction 9.3 Generation of Endothelial-lined Channels 9.4 Perfusion of Endothelial-lined Channels 9.4.1 Media Monitoring Evaporation Cell Metabolism 9.4.2 Inflammatory Cell Stimulation 9.4.3 Whole Blood Perfusion 10 Discussion 10.1 Organ-Disc Technology 10.2 Scalable, Precise and Robust Organ-Disc Fabrication 10.2.1 Fabrication of Thermoplastic Organ-Discs 10.2.2 Fabrication of TPE Modules 10.2.3 Integration of TPE Modules to Organ-Discs 10.3 Tunable, Pump- and Tubing-free Perfusion 10.4 On-Disc Tissue Culture 10.4.1 3D Tissues 10.4.2 Blood Vessel-like Structures 10.4.3 Tissue Characterization and Treatment 10.5 On-Disc Blood Perfusion 11 Summary and Conclusion 12 References 13 Appendi

    Towards micropump- and microneedle-based drug delivery using Micro Transdermal Interface Platforms (MicroTIPs)

    Get PDF
    Micro Transdermal Interface Platforms (MicroTIPs) will combine minimally invasive microneedle arrays with highly miniaturized sensors, actuators, control electronics, wireless communications and artificial intelligence. These patch-like devices will be capable of autonomous physiological monitoring and transdermal drug delivery, resulting in increased patient adherence and devolved healthcare. In this paper, we experimentally demonstrate the feasibility of controlled transdermal drug delivery using a combination of 500 μm tall silicon microneedles, a commercial micropump, pressure and flow sensors, and bespoke electronics. Using ex-vivo human skin samples and a customized application/retraction system, leak-free delivery of volumes ranging from 0.7-1.1 mL has been achieved in under one hour. Clinical Relevance — This work experimentally confirms the feasibility of combining micropumps with microneedle arrays for applications in transdermal drug delivery

    Implantable Microsystem Technologies For Nanoliter-Resolution Inner Ear Drug Delivery

    Get PDF
    Advances in protective and restorative biotherapies have created new opportunities to use site-directed, programmable drug delivery systems to treat auditory and vestibular disorders. Successful therapy development that leverages the transgenic, knock-in, and knock-out variants of mouse models of human disease requires advanced microsystems specifically designed to function with nanoliter precision and with system volumes suitable for implantation. The present work demonstrates a novel biocompatible, implantable, and scalable microsystem consisted of a thermal phase-change peristaltic micropump with wireless control and a refillable reservoir. The micropump is fabricated around a catheter microtubing (250 μm OD, 125 μm ID) that provided a biocompatible leak-free flow path while avoiding complicated microfluidic interconnects. Direct-write micro-scale printing technology was used to build the mechanical components of the pump around the microtubing directly on the back of a printed circuit board assembly. In vitro characterization results indicated nanoliter resolution control over the desired flow rates of 10–100 nL/min by changing the actuation frequency, with negligible deviations in presence of up to 10× greater than physiological backpressures and ±3°C ambient temperature variation. A biocompatibility study was performed to evaluate material suitability for chronic subcutaneous implantation and clinical translational development. A stand-alone, refillable, in-plane, scalable, and fully implantable microreservoir platform was designed and fabricated to be integrated with the micropump. The microreservoir consists two main components: a cavity for storing the drug and a septum for refilling. The cavity membrane is fabricated with thin Parylene-C layers, using a polyethylene glycol (PEG) sacrificial layer. The septum thickness is minimized by pre-compression down to 1 mm. The results of in vitro characterization indicated negligible restoring force for the optimized cavity membrane and thousands of punctures through the septum without leakage. The micropump and microreservoir were integrated into microsystems which were implanted in mice. The microtubing was implanted into the round window membrane niche for infusion of a known ototoxic compound (sodium salicylate) at 50 nL/min for 20 min. Real-time shifts in distortion product otoacoustic emission thresholds and amplitudes were measured during the infusion. The results match with syringe pump gold standard. For the first time a miniature and yet scalable microsystem for inner ear drug delivery was developed, enabling drug discovery opportunities and translation to human

    Rapid Prototyping of Microfluidic Devices:Realization of Magnetic Micropumps, Fuel Cells and Protein Preconcentrators

    Get PDF
    With the growing importance of miniaturized energy applications and the development of micro Total Analysis Systems (μTAS), we have realized microfluidic devices, namely, magnetic micropumps, microfluidic fuel cells and membrane-based protein preconcentrators, all having high application potential in future. The choice of rapid prototyping microfabrication technologies and the selection of affordable materials are important aspects, when thinking of commercialization. Thus, we have employed powder blasting, polymer molding and assembly technologies during devices fabrication throughout the thesis. The first type of microfluidic device that we present is a poly(methyl methacrylate) (PMMA) ball-valve micropump with two different designs of the electromagnetic actuator, as optimized by the finite element method. The integration of a permanent magnet in a flexible polydimethylsiloxane (PDMS) membrane, which is clamped into PMMA structure, is proposed for providing a large stroke of the pumping membrane, making the micropump bubble-tolerant and self-priming Focusing on low power consumption for μTAS integration, another type of magnetic micropump with active valves is realized. It consists of a microfluidic chamber structure in glass that is assembled with a PDMS sheet, which comprises two valving membranes and a central actuation membrane, having each an integrated permanent magnet that is peristaltically actuated by a rotating arc-shaped permanent magnets assembly. A lumped circuit model is developed to predict and describe the frequency-dependent flow rate behavior for this type of pump. Powder blasting and PDMS molding rapid prototyping technologies are employed for realization of these two types of micropumps. Fuel cells with fluid delivery and removal options, having chemical reaction sites and electrode structures that can be realized in a microfluidic format, have high potential for applications. Therefore, microfluidic direct methanol fuel cells with embedded ion- permselective medium are studied and such type of fuel cell is realized by integrating a narrow Nafion strip in a molded elastomeric structure. A mechanical clamping assembly technology enables leakage-free operation and stable performance. The characterization reveals its output power density, using H2O2-based oxidant, is among the high-performance direct methanol fuel cells in microscale. Re-using the technology of the fuel cell chip, with its particular ion-permselective Nafion membrane and assembly method, we also have developed a protein preconcentrator with high purification performance. Our device can preconcentrate negatively charged biomolecules located at the anodic compartment side of the Nafion strip within only a few minutes with a high preconcentration factor. Moreover, a complex microfluidic finite element model is proposed to study and understand the physics of the preconcentration effect. Finally, we conclude the thesis with an outlook on future developments based on our work of the project and on the assembly technologies for microfluidic device integration

    Bridging Flows: Microfluidic End‐User Solutions

    Get PDF

    Projeto e desenvolvimento de minibomba perist?ltica usando medi??o de vaz?o, aplicada a biossensores

    Get PDF
    A minibomba perist?ltica de roletes ? uma boa alternativa para sistemas que requeiram o transporte de fluidos de forma laminar, cont?nua e controlada, como ? o caso do sensoriamento biol?gico baseado na resson?ncia de pl?smons de superf?cie ou SPR (Surface Plasmon Resonance). Nesse sentido, este trabalho apresenta o projeto e desenvolvimento de uma minibomba perist?ltica de roletes, acionada por um motor de passo conectado a uma caixa de redu??o mec?nica, associando-os a programa??o de sua estrutura de acionamento, de modo a possibilitar o ajuste dos par?metros do motor para um controle em malha aberta da vaz?o de sa?da para esses sistemas. Foram desenvolvidos projetos de tr?s diferentes atuadores, com 2, 3 e 4 roletes, usando o software Solidworks em pl?stico ABS, bem como atuadores de nylon com as mesmas caracter?sticas. A implementa??o utilizou uma impressora 3D usando pl?stico ABS para a sua prototipagem, e usinagem para os atuadores de nylon. A estrutura da minibomba de roletes possui dimens?es de 100 mil?metros quadrados em sua superf?cie e 55 mm de altura, sendo realizados de forma otimizadas em fun??o da vaz?o volum?trica. Foram realizados testes com ?gua e ?lcool dilu?do em ?gua (?lcool 20%) com os atuadores de pl?stico ABS, a uma rota??o de 10 a 100 RPM, sendo percebido um melhor comportamento da minibomba ? uma de 80 RPM (sendo esta utilizada para ajuste do motor para os tubos de l?tex e de silicone). Com uso do tubo de l?tex, obteve-se uma vaz?o m?xima, para 2 roletes, de 22,5 ml/min com ?gua e 20,5 ml/min com ?lcool dilu?do na ?gua (?lcool 20%, sendo esta a propor??o usada para todos os casos), para o atuador com 3 roletes, obteve-se 20,5 ml/min com ?gua e 18,5 ml/min com ?lcool dilu?do na ?gua e, para 4 roletes, foi obtido 20,5 ml/min com ?gua e 18,0 ml/min com ?lcool dilu?do em ?gua. Uma segunda estrutura foi montada para a obten??o das vaz?es m?ximas para os atuadores usando o tubo de silicone, obtendo uma vaz?o m?xima, para 2 roletes, de 27,5 ml/min com ?gua e 22,0 ml/min com ?lcool dilu?do em ?gua, com o atuador de 3 roletes, obteve-se 27,5 ml/min com ?gua e 22,5 ml/min com ?lcool dilu?do em ?gua e, para 4 roletes, foi obtido uma vaz?o m?xima de 17,5 ml/min com ?gua e 16,0 ml/min com ?lcool dilu?do em ?gua. Contudo, para uma rota??o superior a 80 RPM, para os dois casos (tubo de l?tex e silicone) n?o foram observadas quaisquer varia??es da vaz?o para nenhum dos prot?tipos. O sistema proposto para mensurar a vaz?o ? composto de um microcontrolador para aquisi??o e processamento dos dados e circuitos de condicionamento do sinal. Na mesma estrutura tamb?m foi desenvolvida uma interface para visualizar e permitir a intera??o com a minibomba no tocante ? indica??o da vaz?o, calibrada em fun??o da rota??o do motor. A interface com o usu?rio ? realizada usando um display tipo LCD e bot?es que permitem indicar a vaz?o desejada. Foram realizados, ainda, testes com a press?o hidrost?tica e sua correla??o com a vaz?o medida. Finalmente, a minibomba perist?ltica de roletes proposta apresenta caracter?sticas dimensionais que possibilitam a portabilidade e um baixo consumo de pot?ncia, pois a fonte utilizada para a alimenta??o da minibomba ? a mesma que alimenta o Arduino, sendo esta uma fonte externa de 12V com uma corrente de 1A, tendo assim uma pot?ncia consumida de 12W

    Magnetically actuated micropumps

    Get PDF
    "Lab-On-a-Chip" (LOC) systems are intended to transpose complete laboratory instrumentations on the few square centimetres of a single microfluidic chip. With such devices the objective is to minimize the time and cost associated with routine biological analysis while improving reproducibility. At the heart of these systems, a fluid delivery unit controls and transfers tiny quantities of liquids enabling the biological assays. This explains the need for robust integrated micropumps as a precondition for the development of many LOC devices. In this context, we have developed a rapid prototyping method for the fabrication of microfluidic chips in plastic and glass materials. The microfabrication principle, which is based on the powder blasting microstructuring process, was used to build devices in either polymethylmethacrylate (PMMA) or borosilicate glass. Various types of micropumps have been developed which were all based on external magnetic actuation. The use of ferrofluids (or magnetic liquids) has been the subject of the first part of the research. A piston pump using a ferrofluid plug moved by an external magnet has been studied. The integration of a rare-earth material (NdFeB) in a flexible polydimethylsiloxane (PDMS) membrane, in the form of a powder or as a classical permanent magnet, has then been proposed. An external electromagnet was used to actuate the magnet-containing diaphragm of a reciprocating micropump. Different types of valves, which constitute the critical element in reciprocating micropumps, have also been investigated. We have studied silicone membrane valves, nozzle-diffuser elements and ball valves. While nozzle-diffuser elements present the simplest valving solution from a manufacturing point of view, ball valves have been proposed as a very promising alternative due to their high efficiency. Together with the detailed characterization of the prototypes, we have proposed analytical models that predict the hydrodynamic behaviour of the micropumps. The performances of our micropumps indicate that magnetic actuation is well adapted for LOC microsystems. While we have demonstrated that our proposed microfabrication technique is an excellent rapid prototyping method for disposable plastic devices, our glass micropumps present a competitive low-cost alternative satisfying criteria of biocompatibility and high temperature (130 °C) resistance

    Usability and Applicability of Microfluidic Cell Culture Systems

    Get PDF
    corecore