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Sample records for microfluidic perfusion system

  1. Multiwell cell culture plate format with integrated microfluidic perfusion system

    NASA Astrophysics Data System (ADS)

    Domansky, Karel; Inman, Walker; Serdy, Jim; Griffith, Linda G.

    2006-01-01

    A new cell culture analog has been developed. It is based on the standard multiwell cell culture plate format but it provides perfused three-dimensional cell culture capability. The new capability is achieved by integrating microfluidic valves and pumps into the plate. The system provides a means to conduct high throughput assays for target validation and predictive toxicology in the drug discovery and development process. It can be also used for evaluation of long-term exposure to drugs or environmental agents or as a model to study viral hepatitis, cancer metastasis, and other diseases and pathological conditions.

  2. Microfluidic perfusion systems for secretion fingerprint analysis of pancreatic islets: applications, challenges and opportunities.

    PubMed

    Castiello, F Rafael; Heileman, Khalil; Tabrizian, Maryam

    2016-02-01

    A secretome signature is a heterogeneous profile of secretions present in a single cell type. From the secretome signature a smaller panel of proteins, namely a secretion fingerprint, can be chosen to feasibly monitor specific cellular activity. Based on a thorough appraisal of the literature, this review explores the possibility of defining and using a secretion fingerprint to gauge the functionality of pancreatic islets of Langerhans. It covers the state of the art regarding microfluidic perfusion systems used in pancreatic islet research. Candidate analytical tools to be integrated within microfluidic perfusion systems for dynamic secretory fingerprint monitoring were identified. These analytical tools include patch clamp, amperometry/voltametry, impedance spectroscopy, field effect transistors and surface plasmon resonance. Coupled with these tools, microfluidic devices can ultimately find applications in determining islet quality for transplantation, islet regeneration and drug screening of therapeutic agents for the treatment of diabetes. PMID:26732665

  3. Simplified prototyping of perfusable polystyrene microfluidics

    PubMed Central

    Tran, Reginald; Ahn, Byungwook; R. Myers, David; Qiu, Yongzhi; Sakurai, Yumiko; Moot, Robert; Mihevc, Emma; Trent Spencer, H.; Doering, Christopher; A. Lam, Wilbur

    2014-01-01

    Cell culture in microfluidic systems has primarily been conducted in devices comprised of polydimethylsiloxane (PDMS) or other elastomers. As polystyrene (PS) is the most characterized and commonly used substrate material for cell culture, microfluidic cell culture would ideally be conducted in PS-based microsystems that also enable tight control of perfusion and hydrodynamic conditions, which are especially important for culture of vascular cell types. Here, we report a simple method to prototype perfusable PS microfluidics for endothelial cell culture under flow that can be fabricated using standard lithography and wet laboratory equipment to enable stable perfusion at shear stresses up to 300 dyn/cm2 and pumping pressures up to 26 kPa for at least 100 h. This technique can also be extended to fabricate perfusable hybrid PS-PDMS microfluidics of which one application is for increased efficiency of viral transduction in non-adherent suspension cells by leveraging the high surface area to volume ratio of microfluidics and adhesion molecules that are optimized for PS substrates. These biologically compatible microfluidic devices can be made more accessible to biological-based laboratories through the outsourcing of lithography to various available microfluidic foundries. PMID:25379106

  4. Development of an Integrated Microfluidic Perfusion Cell Culture System for Real-Time Microscopic Observation of Biological Cells

    PubMed Central

    Lin, Lung; Wang, Shih-Siou; Wu, Min-Hsien; Oh-Yang, Chih-Chin

    2011-01-01

    This study reports an integrated microfluidic perfusion cell culture system consisting of a microfluidic cell culture chip, and an indium tin oxide (ITO) glass-based microheater chip for micro-scale perfusion cell culture, and its real-time microscopic observation. The system features in maintaining both uniform, and stable chemical or thermal environments, and providing a backflow-free medium pumping, and a precise thermal control functions. In this work, the performance of the medium pumping scheme, and the ITO glass microheater were experimentally evaluated. Results show that the medium delivery mechanism was able to provide pumping rates ranging from 15.4 to 120.0 μL·min−1. In addition, numerical simulation and experimental evaluation were conducted to verify that the ITO glass microheater was capable of providing a spatially uniform thermal environment, and precise temperature control with a mild variation of ±0.3 °C. Furthermore, a perfusion cell culture was successfully demonstrated, showing the cultured cells were kept at high cell viability of 95 ± 2%. In the process, the cultured chondrocytes can be clearly visualized microscopically. As a whole, the proposed cell culture system has paved an alternative route to carry out real-time microscopic observation of biological cells in a simple, user-friendly, and low cost manner. PMID:22164082

  5. Control of perfusable microvascular network morphology using a multiculture microfluidic system.

    PubMed

    Whisler, Jordan A; Chen, Michelle B; Kamm, Roger D

    2014-07-01

    The mechanical and biochemical microenvironment influences the morphological characteristics of microvascular networks (MVNs) formed by endothelial cells (ECs) undergoing the process of vasculogenesis. The objective of this study was to quantify the role of individual factors in determining key network parameters in an effort to construct a set of design principles for engineering vascular networks with prescribed morphologies. To achieve this goal, we developed a multiculture microfluidic platform enabling precise control over paracrine signaling, cell-seeding densities, and hydrogel mechanical properties. Human umbilical vein endothelial cells (HUVECs) were seeded in fibrin gels and cultured alongside human lung fibroblasts (HLFs). The engineered vessels formed in our device contained patent, perfusable lumens. Communication between the two cell types was found to be critical in avoiding network regression and maintaining stable morphology beyond 4 days. The number of branches, average branch length, percent vascularized area, and average vessel diameter were found to depend uniquely on several input parameters. Importantly, multiple inputs were found to control any given output network parameter. For example, the vessel diameter can be decreased either by applying angiogenic growth factors--vascular endothelial growth factor (VEGF) and sphingosine-1-phsophate (S1P)--or by increasing the fibrinogen concentration in the hydrogel. These findings introduce control into the design of MVNs with specified morphological properties for tissue-specific engineering applications. PMID:24151838

  6. A pump-free membrane-controlled perfusion microfluidic platform.

    PubMed

    Goral, Vasiliy N; Tran, Elizabeth; Yuen, Po Ki

    2015-09-01

    In this article, we present a microfluidic platform for passive fluid pumping for pump-free perfusion cell culture, cell-based assay, and chemical applications. By adapting the passive membrane-controlled pumping principle from the previously developed perfusion microplate, which utilizes a combination of hydrostatic pressure generated by different liquid levels in the wells and fluid wicking through narrow strips of a porous membrane connecting the wells to generate fluid flow, a series of pump-free membrane-controlled perfusion microfluidic devices was developed and their use for pump-free perfusion cell culture and cell-based assays was demonstrated. Each pump-free membrane-controlled perfusion microfluidic device comprises at least three basic components: an open well for generating fluid flow, a micron-sized deep chamber/channel for cell culture or for fluid connection, and a wettable porous membrane for controlling the fluid flow. Each component is fluidically connected either by the porous membrane or by the micron-sized deep chamber/channel. By adapting and incorporating the passive membrane-controlled pumping principle into microfluidic devices, all the benefits of microfluidic technologies, such as small sample volumes, fast and efficient fluid exchanges, and fluid properties at the micro-scale, can be fully taken advantage of with this pump-free membrane-controlled perfusion microfluidic platform. PMID:26392835

  7. A perfusion-capable microfluidic bioreactor for assessing microbial heterologous protein production.

    PubMed

    Mozdzierz, Nicholas J; Love, Kerry R; Lee, Kevin S; Lee, Harry L T; Shah, Kartik A; Ram, Rajeev J; Love, J Christopher

    2015-07-21

    We present an integrated microfluidic bioreactor for fully continuous perfusion cultivation of suspended microbial cell cultures. This system allowed continuous and stable heterologous protein expression by sustaining the cultivation of Pichia pastoris over 11 days. This technical capability also allowed testing the impact of perfusion conditions on protein expression. This advance should enable small-scale models for process optimization in continuous biomanufacturing. PMID:26055071

  8. A perfusion-capable microfluidic bioreactor for assessing microbial heterologous protein production

    PubMed Central

    Mozdzierz, Nicholas J.; Love, Kerry R.; Lee, Kevin S.; Lee, Harry L. T.; Shah, Kartik A.; Ram, Rajeev J.

    2015-01-01

    We present an integrated microfluidic bioreactor for fully continuous perfusion cultivation of suspended microbial cell cultures. This system allowed continuous and stable heterologous protein expression by sustaining the cultivation of Pichia pastoris over 11 days. This technical capability also allowed testing the impact of perfusion conditions on protein expression. This advance should enable small-scale models for process optimization in continuous biomanufacturing. PMID:26055071

  9. An integrated microfluidic cell culture system for high-throughput perfusion three-dimensional cell culture-based assays: effect of cell culture model on the results of chemosensitivity assays.

    PubMed

    Huang, Song-Bin; Wang, Shih-Siou; Hsieh, Chia-Hsun; Lin, Yung Chang; Lai, Chao-Sung; Wu, Min-Hsien

    2013-03-21

    Although microfluidic cell culture systems are versatile tools for cellular assays, their use has yet to set in motion an evolutionary shift away from conventional cell culture methods. This situation is mainly due to technical hurdles: the operational barriers to the end-users, the lack of compatible detection schemes capable of reading out the results of a microfluidic-based cellular assay, and the lack of fundamental data to bridge the gap between microfluidic and conventional cell culture models. To address these issues, we propose a high-throughput, perfusion, three-dimensional (3-D) microfluidic cell culture system encompassing 30 microbioreactors. This integrated system not only aims to provide a user-friendly cell culture tool for biologists to perform assays but also to enable them to obtain precise data. Its technical features include (i) integration of a heater chip based on transparent indium tin oxide glass, providing stable thermal conditions for cell culturing; (ii) a microscale 3-D culture sample loading scheme that is both efficient and precise; (iii) a non-mechanical pneumatically driven multiplex medium perfusion mechanism; and (iv) a microplate reader-compatible waste medium collector array for the subsequent high throughput bioassays. In this study, we found that the 3-D culture sample loading method provided uniform sample loading [coefficient of variation (CV): 3.2%]. In addition, the multiplex medium perfusion mechanism led to reasonably uniform (CV: 3.6-6.9%) medium pumping rates in the 30 microchannels. Moreover, we used the proposed system to perform a successful cell culture-based chemosensitivity assay. To determine the effects of cell culture models on the cellular proliferation, and the results of chemosensitivity assays, we compared our data with that obtained using three conventional cell culture models. We found that the nature of the cell culture format could lead to different evaluation outcomes. Consequently, when establishing a

  10. A microfluidically perfused three dimensional human liver model.

    PubMed

    Rennert, Knut; Steinborn, Sandra; Gröger, Marko; Ungerböck, Birgit; Jank, Anne-Marie; Ehgartner, Josef; Nietzsche, Sandor; Dinger, Julia; Kiehntopf, Michael; Funke, Harald; Peters, Frank T; Lupp, Amelie; Gärtner, Claudia; Mayr, Torsten; Bauer, Michael; Huber, Otmar; Mosig, Alexander S

    2015-12-01

    Within the liver, non-parenchymal cells (NPCs) are critically involved in the regulation of hepatocyte polarization and maintenance of metabolic function. We here report the establishment of a liver organoid that integrates NPCs in a vascular layer composed of endothelial cells and tissue macrophages and a hepatic layer comprising stellate cells co-cultured with hepatocytes. The three-dimensional liver organoid is embedded in a microfluidically perfused biochip that enables sufficient nutrition supply and resembles morphological aspects of the human liver sinusoid. It utilizes a suspended membrane as a cell substrate mimicking the space of Disse. Luminescence-based sensor spots were integrated into the chip to allow online measurement of cellular oxygen consumption. Application of microfluidic flow induces defined expression of ZO-1, transferrin, ASGPR-1 along with an increased expression of MRP-2 transporter protein within the liver organoids. Moreover, perfusion was accompanied by an increased hepatobiliary secretion of 5(6)-carboxy-2',7'-dichlorofluorescein and an enhanced formation of hepatocyte microvilli. From this we conclude that the perfused liver organoid shares relevant morphological and functional characteristics with the human liver and represents a new in vitro research tool to study human hepatocellular physiology at the cellular level under conditions close to the physiological situation. PMID:26322723

  11. Continuous perfusion microfluidic cell culture array for high-throughput cell-based assays.

    PubMed

    Hung, Paul J; Lee, Philip J; Sabounchi, Poorya; Lin, Robert; Lee, Luke P

    2005-01-01

    We present for the first time a microfluidic cell culture array for long-term cellular monitoring. The 10 x 10 array could potentially assay 100 different cell-based experiments in parallel. The device was designed to integrate the processes used in typical cell culture experiments on a single self-contained microfluidic system. Major functions include repeated cell growth/passage cycles, reagent introduction, and real-time optical analysis. The single unit of the array consists of a circular microfluidic chamber, multiple narrow perfusion channels surrounding the main chamber, and four ports for fluidic access. Human carcinoma (HeLa) cells were cultured inside the device with continuous perfusion of medium at 37 degrees C. The observed doubling time was 1.4 +/- 0.1 days with a peak cell density of approximately 2.5*10(5) cells/cm(2). Cell assay was demonstrated by monitoring the fluorescence localization of calcein AM from 1 min to 10 days after reagent introduction. Confluent cell cultures were passaged within the microfluidic chambers using trypsin and successfully regrown, suggesting a stable culture environment suitable for continuous operation. The cell culture array could offer a platform for a wide range of assays with applications in drug screening, bioinformatics, and quantitative cell biology. PMID:15580587

  12. Experimental Microfluidic System

    NASA Technical Reports Server (NTRS)

    Culbertson, Christopher; Gonda, Steve; Ramsey, John Michael

    2005-01-01

    The ultimate goal of this project is to integrate microfluidic devices with NASA's space bioreactor systems. In such a system, the microfluidic device would provide realtime feedback control of the bioreactor by monitoring pH, glucose, and lactate levels in the cell media; and would provide an analytical capability to the bioreactor in exterrestrial environments for monitoring bioengineered cell products and health changes in cells due to environmental stressors. Such integrated systems could be used as biosentinels both in space and on planet surfaces. The objective is to demonstrate the ability of microfabricated devices to repeatedly and reproducibly perform bead cytometry experiments in micro, lunar, martian, and hypergravity (1.8g).

  13. Perfused drop microfluidic device for brain slice culture-based drug discovery.

    PubMed

    Liu, Jing; Pan, Liping; Cheng, Xuanhong; Berdichevsky, Yevgeny

    2016-06-01

    Living slices of brain tissue are widely used to model brain processes in vitro. In addition to basic neurophysiology studies, brain slices are also extensively used for pharmacology, toxicology, and drug discovery research. In these experiments, high parallelism and throughput are critical. Capability to conduct long-term electrical recording experiments may also be necessary to address disease processes that require protein synthesis and neural circuit rewiring. We developed a novel perfused drop microfluidic device for use with long term cultures of brain slices (organotypic cultures). Slices of hippocampus were placed into wells cut in polydimethylsiloxane (PDMS) film. Fluid level in the wells was hydrostatically controlled such that a drop was formed around each slice. The drops were continuously perfused with culture medium through microchannels. We found that viable organotypic hippocampal slice cultures could be maintained for at least 9 days in vitro. PDMS microfluidic network could be readily integrated with substrate-printed microelectrodes for parallel electrical recordings of multiple perfused organotypic cultures on a single MEA chip. We expect that this highly scalable perfused drop microfluidic device will facilitate high-throughput drug discovery and toxicology. PMID:27194028

  14. Blood Perfusion in Microfluidic Models of Pulmonary Capillary Networks: Role of Geometry and Hematocrit

    NASA Astrophysics Data System (ADS)

    Stauber, Hagit; Waisman, Dan; Sznitman, Josue; Technion-IIT Team; Department of Neonatology Carmel Medical Center; Faculty of Medicine-Technion IIT Collaboration

    2015-11-01

    Microfluidic platforms are increasingly used to study blood microflows at true physiological scale due to their ability to overcome manufacturing obstacle of complex anatomical morphologies, such as the organ-specific architectures of the microcirculation. In the present work, we utilize microfluidic platforms to devise in vitro models of the underlying pulmonary capillary networks (PCN), where capillary lengths and diameters are similar to the size of RBCs (~ 5-10 μm). To better understand flow characteristics and dispersion of red blood cells (RBCs) in PCNs, we have designed microfluidic models of alveolar capillary beds inspired by the seminal ``sheet flow'' model of Fung and Sobin (1969). Our microfluidic PCNs feature confined arrays of staggered pillars with diameters of ~ 5,7 and 10 μm, mimicking the dense structure of pulmonary capillary meshes. The devices are perfused with suspensions of RBCs at varying hematocrit levels under different flow rates. Whole-field velocity patterns using micro-PIV and single-cell tracking using PTV are obtained with fluorescently-labelled RBCs and discussed. Our experiments deliver a real-scale quantitative description of RBC perfusion characteristics across the pulmonary capillary microcirculation.

  15. A microfluidic perfusion platform for cultivation and screening study of motile microalgal cells

    PubMed Central

    Eu, Young-Jae; Park, Hye-Sun; Kim, Dong-Pyo; Wook Hong, Jong

    2014-01-01

    Systematic screening of algal cells is getting huge interest due to their capability of producing lipid-based biodiesel. Here, we introduce a new microfluidic platform composed of an array of perfusion chambers designed for long-term cultivation and preliminary screening of motile microalgal cells through loading and releasing of cells to and from the chambers. The chemical environment in each perfusion chamber was independently controlled for 5 days. The effect of nitrogen-depletion on the lipid production, phototaxis behavior in the absence of Ca2+, and cytotoxic effect of herbicide on microalgal cells was successfully monitored and compared with simultaneous control experiments on the platform. The present methodology could be extended to effective screening of algal cells and various cell lines for the production of biodiesel and other useful chemicals. PMID:24803962

  16. Ice matrix in reconfigurable microfluidic systems

    NASA Astrophysics Data System (ADS)

    Bossi, A. M.; Vareijka, M.; Piletska, E. V.; Turner, A. P. F.; Meglinski, I.; Piletsky, S. A.

    2013-07-01

    Microfluidic devices find many applications in biotechnologies. Here, we introduce a flexible and biocompatible microfluidic ice-based platform with tunable parameters and configuration of microfluidic patterns that can be changed multiple times during experiments. Freezing and melting of cavities, channels and complex relief structures created and maintained in the bulk of ice by continuous scanning of an infrared laser beam are used as a valve action in microfluidic systems. We demonstrate that pre-concentration of samples and transport of ions and dyes through the open channels created can be achieved in ice microfluidic patterns by IR laser-assisted zone melting. The proposed approach can be useful for performing separation and sensing processes in flexible reconfigurable microfluidic devices.

  17. Microfluidic Systems for Biosensing

    PubMed Central

    Liu, Kuo-Kang; Wu, Ren-Guei; Chuang, Yun-Ju; Khoo, Hwa Seng; Huang, Shih-Hao; Tseng, Fan-Gang

    2010-01-01

    In the past two decades, Micro Fluidic Systems (MFS) have emerged as a powerful tool for biosensing, particularly in enriching and purifying molecules and cells in biological samples. Compared with conventional sensing techniques, distinctive advantages of using MFS for biomedicine include ultra-high sensitivity, higher throughput, in-situ monitoring and lower cost. This review aims to summarize the recent advancements in two major types of micro fluidic systems, continuous and discrete MFS, as well as their biomedical applications. The state-of-the-art of active and passive mechanisms of fluid manipulation for mixing, separation, purification and concentration will also be elaborated. Future trends of using MFS in detection at molecular or cellular level, especially in stem cell therapy, tissue engineering and regenerative medicine, are also prospected. PMID:22163570

  18. Long term perfusion system supporting adipogenesis

    PubMed Central

    Abbott, Rosalyn D.; Raja, Waseem K.; Wang, Rebecca Y.; Stinson, Jordan A.; Glettig, Dean L.; Burke, Kelly A.; Kaplan, David L.

    2015-01-01

    Adipose tissue engineered models are needed to enhance our understanding of disease mechanisms and for soft tissue regenerative strategies. Perfusion systems generate more physiologically relevant and sustainable adipose tissue models, however adipocytes have unique properties that make culturing them in a perfusion environment challenging. In this paper we describe the methods involved in the development of two perfusion culture systems (2D and 3D) to test their applicability for long term in vitro adipogenic cultures. It was hypothesized that a silk protein biomaterial scaffold would provide a 3D framework, in combination with perfusion flow, to generate a more physiologically relevant sustainable adipose tissue engineered model than 2D cell culture. Consistent with other studies evaluating 2D and 3D culture systems for adipogenesis we found that both systems successfully model adipogensis, however 3D culture systems were more robust, providing the mechanical structure required to contain the large, fragile adipocytes that were lost in 2D perfused culture systems. 3D perfusion also stimulated greater lipogenesis and lipolysis and resulted in decreased secretion of LDH compared to 2D perfusion. Regardless of culture configuration (2D or 3D) greater glycerol was secreted with the increased nutritional supply provided by perfusion of fresh media. These results are promising for adipose tissue engineering applications including long term cultures for studying disease mechanisms and regenerative approaches, where both acute (days to weeks) and chronic (weeks to months) cultivation are critical for useful insight. PMID:25843606

  19. Modular microfluidic system for biological sample preparation

    SciTech Connect

    Rose, Klint A.; Mariella, Jr., Raymond P.; Bailey, Christopher G.; Ness, Kevin Dean

    2015-09-29

    A reconfigurable modular microfluidic system for preparation of a biological sample including a series of reconfigurable modules for automated sample preparation adapted to selectively include a) a microfluidic acoustic focusing filter module, b) a dielectrophoresis bacteria filter module, c) a dielectrophoresis virus filter module, d) an isotachophoresis nucleic acid filter module, e) a lyses module, and f) an isotachophoresis-based nucleic acid filter.

  20. Self-contained microfluidic systems: a review.

    PubMed

    Boyd-Moss, Mitchell; Baratchi, Sara; Di Venere, Martina; Khoshmanesh, Khashayar

    2016-08-16

    Microfluidic systems enable rapid diagnosis, screening and monitoring of diseases and health conditions using small amounts of biological samples and reagents. Despite these remarkable features, conventional microfluidic systems rely on bulky expensive external equipment, which hinders their utility as powerful analysis tools outside of research laboratories. 'Self-contained' microfluidic systems, which contain all necessary components to facilitate a complete assay, have been developed to address this limitation. In this review, we provide an in-depth overview of self-contained microfluidic systems. We categorise these systems based on their operating mechanisms into three major groups: passive, hand-powered and active. Several examples are provided to discuss the structure, capabilities and shortcomings of each group. In particular, we discuss the self-contained microfluidic systems enabled by active mechanisms, due to their unique capability for running multi-step and highly controllable diagnostic assays. Integration of self-contained microfluidic systems with the image acquisition and processing capabilities of smartphones, especially those equipped with accessory optical components, enables highly sensitive and quantitative assays, which are discussed. Finally, the future trends and possible solutions to expand the versatility of self-contained, stand-alone microfluidic platforms are outlined. PMID:27425637

  1. Microfluidic microarray systems and methods thereof

    DOEpatents

    West, Jay A. A.; Hukari, Kyle W.; Hux, Gary A.

    2009-04-28

    Disclosed are systems that include a manifold in fluid communication with a microfluidic chip having a microarray, an illuminator, and a detector in optical communication with the microarray. Methods for using these systems for biological detection are also disclosed.

  2. Magnetoactive Sponges for Dynamic Control of Microfluidic Flow Patterns in Microphysiological Systems

    PubMed Central

    Yen, Ringo; Chan, Hon Fai; Leong, Kam W; Truskey, George A; Zhao, Xuanhe

    2014-01-01

    We developed a microfluidic flow-control system capable of dynamically generating various flow patterns on demand. The flow-control system is based on novel magnetoactive sponges embedded in microfluidic flow channels. Applying a non-uniform magnetic field compresses the magnetoactive sponge, significantly reducing porosity and hydraulic conductivity. Tuning the applied magnetic field can dynamically vary the flow rate in the microfluidic channel. Pulsatile and physiological flow patterns with frequency between 1 and 3 Hz, flow rates between 0.5 and 10 μL/min and duration over 3 weeks have been achieved. Smooth muscle cells in engineered blood vessels perfused for 7 days aligned perpendicular to the flow direction under pulsatile but not steady flow, similar to the in vivo orientation. Owing to its various advantages over traditional flow-control methods, the new system potentially has important applications in microfluidic-based microphysiological systems to simulate the physiological nature of blood flow. PMID:24310854

  3. Bio-functionalized silk hydrogel microfluidic systems.

    PubMed

    Zhao, Siwei; Chen, Ying; Partlow, Benjamin P; Golding, Anne S; Tseng, Peter; Coburn, Jeannine; Applegate, Matthew B; Moreau, Jodie E; Omenetto, Fiorenzo G; Kaplan, David L

    2016-07-01

    Bio-functionalized microfluidic systems were developed based on a silk protein hydrogel elastomeric materials. A facile multilayer fabrication method using gelatin sacrificial molding and layer-by-layer assembly was implemented to construct interconnected, three dimensional (3D) microchannel networks in silk hydrogels at 100 μm minimum feature resolution. Mechanically activated valves were implemented to demonstrate pneumatic control of microflow. The silk hydrogel microfluidics exhibit controllable mechanical properties, long-term stability in various environmental conditions, tunable in vitro and in vivo degradability in addition to optical transparency, providing unique features for cell/tissue-related applications than conventional polydimethylsiloxane (PDMS) and existing hydrogel-based microfluidic options. As demonstrated in the work here, the all aqueous-based fabrication process at ambient conditions enabled the incorporation of active biological substances in the bulk phase of these new silk microfluidic systems during device fabrication, including enzymes and living cells, which are able to interact with the fluid flow in the microchannels. These silk hydrogel-based microfluidic systems offer new opportunities in engineering active diagnostic devices, tissues and organs that could be integrated in vivo, and for on-chip cell sensing systems. PMID:27077566

  4. Perfusion and Characterization of an Endothelial Cell-Seeded Modular Tissue Engineered Construct Formed in a Microfluidic Remodeling Chamber

    PubMed Central

    Khan, Omar F.

    2010-01-01

    Tissue engineered constructs containing tortuous endothelial cell-lined perfusion channels were formed by randomly assembling endothelial cell-seeded submillimeter-sized collagen cylinders (modules) into a microfluidic perfusion chamber. The interconnected void space produced by random module packing created flow channels that were lined with endothelial cells. The effect of perfusion (0.5 mL min−1, Re* = 14.36 and shear stress = 0.64 dyn cm−2) through the tortuous channels on construct remodeling and endothelium quiescence was studied. Over time, modules fused at their points of contact and as they contracted, decreased the internal void space, which reduced the overall perfusion through the construct. As compared to static controls, perfusion caused a transient increase in activation (ICAM-1 and VCAM-1 expression) after 1 hour followed by a decrease after 24 hours. Proliferation (by BrdU) was reduced significantly, while KLF2, which is upregulated with atheroprotective laminar shear stress, was upregulated significantly after 24 hours. VE-cadherin became discontinuous and was significantly downregulated after 24 hours, which was likely caused by the dismantling of the endothelial cell adherens junctions during remodeling. Collectively, these outcomes suggest that flow through the construct did not drive the endothelial cells towards an inflamed, “atherosclerotic like” disturbed flow pathology. PMID:20678792

  5. Polymer-based platform for microfluidic systems

    DOEpatents

    Benett, William; Krulevitch, Peter; Maghribi, Mariam; Hamilton, Julie; Rose, Klint; Wang, Amy W.

    2009-10-13

    A method of forming a polymer-based microfluidic system platform using network building blocks selected from a set of interconnectable network building blocks, such as wire, pins, blocks, and interconnects. The selected building blocks are interconnectably assembled and fixedly positioned in precise positions in a mold cavity of a mold frame to construct a three-dimensional model construction of a microfluidic flow path network preferably having meso-scale dimensions. A hardenable liquid, such as poly (dimethylsiloxane) is then introduced into the mold cavity and hardened to form a platform structure as well as to mold the microfluidic flow path network having channels, reservoirs and ports. Pre-fabricated elbows, T's and other joints are used to interconnect various building block elements together. After hardening the liquid the building blocks are removed from the platform structure to make available the channels, cavities and ports within the platform structure. Microdevices may be embedded within the cast polymer-based platform, or bonded to the platform structure subsequent to molding, to create an integrated microfluidic system. In this manner, the new microfluidic platform is versatile and capable of quickly generating prototype systems, and could easily be adapted to a manufacturing setting.

  6. Cardiac tissue engineering using perfusion bioreactor systems

    PubMed Central

    Radisic, Milica; Marsano, Anna; Maidhof, Robert; Wang, Yadong; Vunjak-Novakovic, Gordana

    2009-01-01

    This protocol describes tissue engineering of synchronously contractile cardiac constructs by culturing cardiac cell populations on porous scaffolds (in some cases with an array of channels) and bioreactors with perfusion of culture medium (in some cases supplemented with an oxygen carrier). The overall approach is ‘biomimetic’ in nature as it tends to provide in vivo-like oxygen supply to cultured cells and thereby overcome inherent limitations of diffusional transport in conventional culture systems. In order to mimic the capillary network, cells are cultured on channeled elastomer scaffolds that are perfused with culture medium that can contain oxygen carriers. The overall protocol takes 2–4 weeks, including assembly of the perfusion systems, preparation of scaffolds, cell seeding and cultivation, and on-line and end-point assessment methods. This model is well suited for a wide range of cardiac tissue engineering applications, including the use of human stem cells, and high-fidelity models for biological research. PMID:18388955

  7. Acoustically-driven microfluidic systems

    SciTech Connect

    Wang, A W; Benett, W J; Tarte, L R

    2000-06-23

    We have demonstrated a non-contact method of concentrating and mixing particles in a plastic microfluidic chamber employing acoustic radiation pressure. A flaw cell package has also been designed that integrates liquid sample interconnects, electrical contacts and a removable sample chamber. Experiments were performed on 1, 3, 6, and 10 {micro}m polystyrene beads. Increased antibody binding to a solid-phase substrate was observed in the presence of acoustic mixing due to improve mass transport.

  8. Integrated perfusion and separation systems for entrainment of insulin secretion from islets of Langerhans.

    PubMed

    Yi, Lian; Wang, Xue; Dhumpa, Raghuram; Schrell, Adrian M; Mukhitov, Nikita; Roper, Michael G

    2015-02-01

    A microfluidic system was developed to investigate the entrainment of insulin secretion from islets of Langerhans to oscillatory glucose levels. A gravity-driven perfusion system was integrated with a microfluidic system to deliver sinusoidal glucose waveforms to the islet chamber. Automated manipulation of the height of the perfusion syringes allowed precise control of the ratio of two perfusion solutions into a chamber containing 1-10 islets. Insulin levels in the perfusate were measured using an online competitive electrophoretic immunoassay with a sampling period of 10 s. The insulin immunoassay had a detection limit of 3 nM with RSDs of calibration points ranging from 2-8%. At 11 mM glucose, insulin secretion from single islets was oscillatory with a period ranging from 3-6 min. Application of a small amplitude sinusoidal wave of glucose with a period of 5 or 10 min, shifted the period of the insulin oscillations to this forcing period. Exposing groups of 6-10 islets to a sinusoidal glucose wave synchronized their behavior, producing a coherent pulsatile insulin response from the population. These results demonstrate the feasibility of the developed system for the study of oscillatory insulin secretion and can be easily modified for investigating the dynamic nature of other hormones released from different cell types. PMID:25474044

  9. Microfluidic systems for single DNA dynamics

    PubMed Central

    Mai, Danielle J.; Brockman, Christopher

    2012-01-01

    Recent advances in microfluidics have enabled the molecular-level study of polymer dynamics using single DNA chains. Single polymer studies based on fluorescence microscopy allow for the direct observation of non-equilibrium polymer conformations and dynamical phenomena such as diffusion, relaxation, and molecular stretching pathways in flow. Microfluidic devices have enabled the precise control of model flow fields to study the non-equilibrium dynamics of soft materials, with device geometries including curved channels, cross-slots, and microfabricated obstacles and structures. This review explores recent microfluidic systems that have advanced the study of single polymer dynamics, while identifying new directions in the field that will further elucidate the relationship between polymer microstructure and bulk rheological properties. PMID:23139700

  10. Recent advances in microfluidic detection systems

    PubMed Central

    Baker, Christopher A; Duong, Cindy T; Grimley, Alix; Roper, Michael G

    2009-01-01

    There are numerous detection methods available for methods are being put to use for detection on these miniaturized systems, with the analyte of interest driving the choice of detection method. In this article, we summarize microfluidic 2 years. More focus is given to unconventional approaches to detection routes and novel strategies for performing high-sensitivity detection. PMID:20414455

  11. Microfluidic-Based Robotic Sampling System for Radioactive Solutions

    SciTech Connect

    Jack D. Law; Julia L. Tripp; Tara E. Smith; Veronica J. Rutledge; Troy G. Garn; John Svoboda; Larry Macaluso

    2014-02-01

    A novel microfluidic based robotic sampling system has been developed for sampling and analysis of liquid solutions in nuclear processes. This system couples the use of a microfluidic sample chip with a robotic system designed to allow remote, automated sampling of process solutions in-cell and facilitates direct coupling of the microfluidic sample chip with analytical instrumentation. This system provides the capability for near real time analysis, reduces analytical waste, and minimizes the potential for personnel exposure associated with traditional sampling methods. A prototype sampling system was designed, built and tested. System testing demonstrated operability of the microfluidic based sample system and identified system modifications to optimize performance.

  12. Fundamental principles and applications of microfluidic systems.

    PubMed

    Ong, Soon-Eng; Zhang, Sam; Du, Hejun; Fu, Yongqing

    2008-01-01

    Microelectromechanical systems (MEMS) technology has provided the platform for the miniaturization of analytical devices for biological applications. Beside the fabrication technology, the study and understanding of flow characteristics of fluid in micrometer or even nanometer scale is vital for the successful implementation of such miniaturized systems. Microfluidics is currently under the spotlight for medical diagnostics and many other bio-analysis as its physical size manifested numerous advantages over lab-based devices. In this review, elementary concepts of fluid and its flow characteristics together with various transport processes and microchannel condition are presented. They are among the fundamental building block for the success in microfluidic systems. Selected application examples include biological cell handling employing different schemes of manipulation and DNA amplification using different microreactor arrangement and fluid flow regime. PMID:17981751

  13. Microfluidic Biosensing Systems Using Magnetic Nanoparticles

    PubMed Central

    Giouroudi, Ioanna; Keplinger, Franz

    2013-01-01

    In recent years, there has been rapidly growing interest in developing hand held, sensitive and cost-effective on-chip biosensing systems that directly translate the presence of certain bioanalytes (e.g., biomolecules, cells and viruses) into an electronic signal. The impressive and rapid progress in micro- and nanotechnology as well as in biotechnology enables the integration of a variety of analytical functions in a single chip. All necessary sample handling and analysis steps are then performed within the chip. Microfluidic systems for biomedical analysis usually consist of a set of units, which guarantees the manipulation, detection and recognition of bioanalytes in a reliable and flexible manner. Additionally, the use of magnetic fields for performing the aforementioned tasks has been steadily gaining interest. This is because magnetic fields can be well tuned and applied either externally or from a directly integrated solution in the biosensing system. In combination with these applied magnetic fields, magnetic nanoparticles are utilized. Some of the merits of magnetic nanoparticles are the possibility of manipulating them inside microfluidic channels by utilizing high gradient magnetic fields, their detection by integrated magnetic microsensors, and their flexibility due to functionalization by means of surface modification and specific binding. Their multi-functionality is what makes them ideal candidates as the active component in miniaturized on-chip biosensing systems. In this review, focus will be given to the type of biosening systems that use microfluidics in combination with magnetoresistive sensors and detect the presence of bioanalyte tagged with magnetic nanoparticles. PMID:24022689

  14. Self-driven perfusion culture system using a paper-based double-layered scaffold.

    PubMed

    Ozaki, Ai; Arisaka, Yoshinori; Takeda, Naoya

    2016-01-01

    Shear stress caused by fluid flow is known to promote tissue development from cells in vivo. Therefore, perfusion cultures have been studied to investigate the mechanisms involved and to fabricate engineered tissues in vitro, particularly those that include blood vessels. Microfluidic devices, which function with fine machinery of chambers and microsyringes for fluid flow and have small culture areas, are conventionally used for perfusion culture. In contrast, we have developed a self-driven perfusion culture system by using a paper-based double-layered scaffold as the fundamental component. Gelatin microfibers were electrospun onto a paper material to prepare the scaffold system, in which the constant perfusion of the medium and the scaffold for cell adhesion/proliferation were functionally divided into a paper and a gelatin microfiber layer, respectively. By applying both the capillary action and siphon phenomenon of the paper-based scaffold, which bridged two medium chambers at different height levels, a self-driven medium flow was achieved and the flow rate was also stable, constant, and quantitatively controllable. Moreover, the culture area was enlargeable to the cm(2) scale. The endothelial cells cultivated on this system oriented along the medium-flow direction, suggesting that the shear stress caused by medium flow was effectively applied. This perfusion culture system is expected to be useful for fabricating three-dimensional and large engineered tissues in the future. PMID:27550929

  15. Microfluidic Impedimetric Cell Regeneration Assay to Monitor the Enhanced Cytotoxic Effect of Nanomaterial Perfusion

    PubMed Central

    Rothbauer, Mario; Praisler, Irene; Docter, Dominic; Stauber, Roland H.; Ertl, Peter

    2015-01-01

    In the last decade, the application of nanomaterials (NMs) in technical products and biomedicine has become a rapidly increasing market trend. As the safety and efficacy of NMs are of utmost importance, new methods are needed to study the dynamic interactions of NMs at the nano-biointerface. However, evaluation of NMs based on standard and static cell culture end-point detection methods does not provide information on the dynamics of living biological systems, which is crucial for the understanding of physiological responses. To bridge this technological gap, we here present a microfluidic cell culture system containing embedded impedance microsensors to continuously and non-invasively monitor the effects of NMs on adherent cells under varying flow conditions. As a model, the impact of silica NMs on the vitality and regenerative capacity of human lung cells after acute and chronic exposure scenarios was studied over an 18-h period following a four-hour NM treatment. Results of the study demonstrated that the developed system is applicable to reliably analyze the consequences of dynamic NM exposure to physiological cell barriers in both nanotoxicology and nanomedicine. PMID:26633532

  16. Microfluidic Impedimetric Cell Regeneration Assay to Monitor the Enhanced Cytotoxic Effect of Nanomaterial Perfusion.

    PubMed

    Rothbauer, Mario; Praisler, Irene; Docter, Dominic; Stauber, Roland H; Ertl, Peter

    2015-12-01

    In the last decade, the application of nanomaterials (NMs) in technical products and biomedicine has become a rapidly increasing market trend. As the safety and efficacy of NMs are of utmost importance, new methods are needed to study the dynamic interactions of NMs at the nano-biointerface. However, evaluation of NMs based on standard and static cell culture end-point detection methods does not provide information on the dynamics of living biological systems, which is crucial for the understanding of physiological responses. To bridge this technological gap, we here present a microfluidic cell culture system containing embedded impedance microsensors to continuously and non-invasively monitor the effects of NMs on adherent cells under varying flow conditions. As a model, the impact of silica NMs on the vitality and regenerative capacity of human lung cells after acute and chronic exposure scenarios was studied over an 18-h period following a four-hour NM treatment. Results of the study demonstrated that the developed system is applicable to reliably analyze the consequences of dynamic NM exposure to physiological cell barriers in both nanotoxicology and nanomedicine. PMID:26633532

  17. Electrostatic actuators for portable microfluidic systems

    NASA Astrophysics Data System (ADS)

    Tice, Joshua

    Both developed and developing nations have an urgent need to diagnose disease cheaply, reliably, and independently of centralized facilities. Microfulidic platforms are well-positioned to address the need for portable diagnostics, mainly due to their obvious advantage in size. However, most microfluidic methods rely on equipment outside of the chip either for driving fluid flow (e.g., syringe pumps) or for taking measurements (e.g., lasers or microscopes). The energy and space requirements of the whole system inhibit portability and contribute to costs. To capitalize on the strengths of microfluidic platforms and address the serious needs of society, system components need to be miniaturized. Also, miniaturization should be accomplished as simply as possible, considering that simplicity is usually requisite for achieving truly transformative technology. Herein, I attempt to address the issue of controlling fluid flow in portable microfluidic systems. I focus on systems that are driven by elastomer-based membrane valves, since these valves are inherently simple, yet they are capable of sophisticated fluid manipulation. Others have attempted to modify pneumatic microvalves for portable applications, e.g., by transitioning to electromagnetic, thermopneumatic, or piezoelectric actuation principles. However, none of these strategies maintain the proper balance of simplicity, functionality, and ease of integration. My research centers on electrostatic actuators, due to their conceptual simplicity and the efficacy of electrostatic forces on the microscale. To ensure easy integration with polymer-based systems, and to maintain simplicity in the fabrication procedure, the actuators were constructed solely from poly(dimethylsiloxane) and multi-walled carbon nanotubes. In addition, the actuators were fabricated exclusively with soft-lithographic techniques. A mathematical model was developed to identify actuator parameters compatible with soft-lithography, and also to

  18. Cell-based bioassays in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Itle, Laura J.; Zguris, Jeanna C.; Pishko, Michael V.

    2004-12-01

    The development of cell-based bioassays for high throughput drug screening or the sensing of biotoxins is contingent on the development of whole cell sensors for specific changes in intracellular conditions and the integration of those systems into sample delivery devices. Here we show the feasibility of using a 5-(and-6)-carboxy SNARF-1, acetoxymethyl ester, acetate, a fluorescent dye capable of responding to changes in intracellular pH, as a detection method for the bacterial endotoxin, lipopolysaccharide. We used photolithography to entrap cells with this dye within poly(ethylene) glyocol diacrylate hydrogels in microfluidic channels. After 18 hours of exposure to lipopolysaccharide, we were able to see visible changes in the fluorescent pattern. This work shows the feasibility of using whole cell based biosensors within microfluidic networks to detect cellular changes in response to exogenous agents.

  19. Structure and fabrication details of an integrated modularized microfluidic system.

    PubMed

    Tian, Qingchang; Mu, Ying; Xu, Yanan; Song, Qi; Yu, Bingwen; Ma, Congcong; Jin, Wei; Jin, Qinhan

    2015-12-01

    This article contains schemes, original experimental data and figures for an integrated modularized microfluidic system described in "An integrated microfluidic system for bovine DNA purification and digital PCR detection [1]". In this data article, we described the structure and fabrication of the integrated modularized microfluidic system. This microfluidic system was applied to isolate DNA from ovine tissue lysate and detect the bovine DNA with digital PCR (dPCR). The DNA extraction efficiency of the microdevice was compared with the efficiency of benchtop protocol. PMID:26594657

  20. 3D functional and perfusable microvascular networks for organotypic microfluidic models.

    PubMed

    Bersini, Simone; Moretti, Matteo

    2015-05-01

    The metastatic dissemination of cancer cells from primary tumors to secondary loci is a complex and multistep process including local invasion, intravasation, survival in the blood stream and extravasation towards the metastatic site. It is well known cancer metastases follow organ-specific pathways with selected primary tumors mainly metastasizing towards a specific panel of secondary organs (Steven Paget's theory 1889). However, circulatory patterns and microarchitecture of capillary networks play a key role in the metastatic spread as well (James Ewing's theory 1929). Taking into account both these factors would be critical to develop more complex and physiologically relevant in vitro cancer models. This review presents recent advances in the generation of microvascularized systems through microfluidic approaches and discusses promising results achieved by organ-on-a-chip platforms mimicking the pathophysiology of the functional units of specific organs. The combination of physiologically-like microvascular networks and organotypic microenvironments would foster a new generation of in vitro cancer models to more effectively screen new therapeutics, design personalized medicine treatments and investigate molecular pathways involved in cancer metastases. PMID:25893395

  1. Real-time monitoring system for microfluidics

    NASA Astrophysics Data System (ADS)

    Sapuppo, F.; Cantelli, G.; Fortuna, L.; Arena, P.; Bucolo, M.

    2007-05-01

    A new non-invasive real-time system for the monitoring and control of microfluidodynamic phenomena is proposed. The general purpose design of such system is suitable for in vitro and in vivo experimental setup and therefore for microfluidic application in the biomedical field such as lab-on-chip and for research studies in the field of microcirculation. The system consists of an ad hoc optical setup for image magnification providing images suitable for image acquisition and processing. The optic system was designed and developed using discrete opto-mechanic components mounted on a breadboard in order to provide an optic path accessible at any point where the information needs to be acquired. The optic sensing, acquisition, and processing were performed using an integrated vision system based on the Cellular Nonlinear Networks (CNNs) analogic technology called Focal Plane Processor (FPP, Eye-RIS, Anafocus) and inserted in the optic path. Ad hoc algorithms were implemented for the real-time analysis and extraction of fluido-dynamic parameters in micro-channels. They were tested on images recorded during in vivo microcirculation experiments on hamsters and then they were applied on images optically acquired and processed in real-time during in vitro experiments on a continuous microfluidic device (serpentine mixer, ThinXXS) with a two-phase fluid.

  2. A Phantom Tissue System for the Calibration of Perfusion Measurements

    PubMed Central

    Mudaliar, Ashvinikumar V.; Ellis, Brent E.; Ricketts, Patricia L.; Lanz, Otto I.; Scott, Elaine P.; Diller, Thomas E.

    2008-01-01

    A convenient method for testing and calibrating surface perfusion sensors has been developed. A phantom tissue model is used to simulate the nondirectional blood flow of tissue perfusion. A computational fluid dynamics (CFD) model was constructed in Fluent® to design the phantom tissue and validate the experimental results. The phantom perfusion system was used with a perfusion sensor based on clearance of thermal energy. A heat flux gage measures the heat flux response of tissue when a thermal event (convective cooling) is applied. The blood perfusion and contact resistance are estimated by a parameter estimation code. From the experimental and analytical results, it was concluded that the probe displayed good measurement repeatability and sensitivity. The experimental perfusion measurements in the tissue were in good agreement with those of the CFD models and demonstrated the value of the phantom tissue system. PMID:19045509

  3. Microfluidic Systems for Studying Neurotransmitters and Neurotransmission

    PubMed Central

    Croushore, Callie A.; Sweedler, Jonathan V.

    2013-01-01

    Neurotransmitters and neuromodulators are molecules within the nervous system that play key roles in cell-to-cell communication. Upon stimulation, neurons release these signaling molecules, which then act at local or distant locations to elicit a physiological response. Ranging from small molecules, such as diatomic gases and amino acids, to larger peptides, these chemical messengers are involved in many functional processes including growth, reproduction, memory and behavior. Understanding signaling molecules and the conditions that govern their release in healthy or damaged networks promises to deliver insights into neural network formation and function. Microfluidic devices can provide optimal cell culture conditions, reduced volume systems, and precise control over the chemical and physical nature of the extracellular environment, making them well-suited for studying neurotransmission and other forms of cell-to-cell signaling. Here we review selected microfluidic approaches that are suitable for monitoring cell-to-cell signaling molecules. We highlight devices that improve in vivo sample collection as well as compartmentalized devices designed to isolate individual neurons or co-cultures in vitro, including a focus on systems used for studying neural injury and regeneration, and devices that allow selective chemical stimulations and the characterization of released molecules. PMID:23474943

  4. Imaging of oxygen in microreactors and microfluidic systems

    NASA Astrophysics Data System (ADS)

    Sun, Shiwen; Ungerböck, Birgit; Mayr, Torsten

    2015-09-01

    This review gives an overview on the state-of-the-art of oxygen imaging in microfluidics. Oxygen imaging using optical oxygen sensors based on luminescence is a versatile and powerful tool for obtaining profoundly space-resolved information of oxygen in microreactors and microfluidic systems. We briefly introduce the principle of oxygen imaging and present techniques of oxygen imaging applied in microreactors and microfluidic devices, including selection criteria and demands of sensing material and basic set-up for a 2D oxygen sensing system. A detailed review of oxygen imaging in microreactors and microfluidic systems is given on different applications in oxygen gradient monitoring, cell culturing, single-cell analysis and chemical reactions. Finally, we discuss challenges and trends of oxygen imaging in microfluidic systems.

  5. Integrated Multi-process Microfluidic Systems for Automating Analysis

    PubMed Central

    Yang, Weichun; Woolley, Adam T.

    2010-01-01

    Microfluidic technologies have been applied extensively in rapid sample analysis. Some current challenges for standard microfluidic systems are relatively high detection limits, and reduced resolving power and peak capacity compared to conventional approaches. The integration of multiple functions and components onto a single platform can overcome these separation and detection limitations of microfluidics. Multiplexed systems can greatly increase peak capacity in multidimensional separations and can increase sample throughput by analyzing many samples simultaneously. On-chip sample preparation, including labeling, preconcentration, cleanup and amplification, can all serve to speed up and automate processes in integrated microfluidic systems. This paper summarizes advances in integrated multi-process microfluidic systems for automated analysis, their benefits and areas for needed improvement. PMID:20514343

  6. Microfluidic System for Automated Cell-based Assays.

    PubMed

    Lee, Philip J; Ghorashian, Navid; Gaige, Terry A; Hung, Paul J

    2007-12-01

    Microfluidic cell culture is a promising technology for applications in the drug screening industry. Key benefits include improved biological function, higher quality cell-based data, reduced reagent consumption, and lower cost. In this work, we demonstrate how a microfluidic cell culture design was adapted to be compatible with the standard 96-well plate format. Key design features include the elimination of tubing and connectors, the ability to maintain long term continuous perfusion cell culture using a passive gravity driven pump, and direct analysis on the outlet wells of the microfluidic plate. A single microfluidic culture plate contained 8 independent flow units, each with 10(4) cells at a flow rate of 50 μl/day (6 minute residence time). The cytotoxicity of the anti-cancer drug etoposide was measured on HeLa cells cultured in this format, using a commercial lactate dehydrogenase (LDH) plate reader assay. The integration of microfluidic cell culture methods with commercial automation capabilities offers an exciting opportunity for improved cell-based screening. PMID:18172509

  7. Dynamical systems techniques for enhancing microfluidic mixing

    NASA Astrophysics Data System (ADS)

    Balasuriya, Sanjeeva

    2015-09-01

    Achieving rapid mixing is often desirable in microfluidic devices, for example in improving reation rates in biotechnological assays. Enhancing mixing within a particular context is often achieved by introducing problem-specific strategies such as grooved or twisted channels, ac electromagnetic fields or oscillatory microsyringe flows. Evaluating the efficiency of these methods is challenging since either experimental fabrication and sensing, or computationally expensive direct numerical simulations with complicated boundary conditions, are required. A review of how mixing can be quantified when velocity fields have been obtained from such situations is presented. A less-known alternative to these methods is offered by dynamical systems, which characterizes the motion of collective fluid parcel trajectories by studying crucial interior flow barriers which move unsteadily, but nevertheless strongly govern mixing possibilities. The methodology behind defining these barriers and quantifying the fluid transport influenced by them is explained. Their application towards several microfluidic situations (e.g. best cross-flow positioning in cross-channel micromixers, usage of channel curvature to enhance mixing within microdroplets traveling in a channel, optimum frequencies of velocity agitations to use) is discussed.

  8. Integrated microfluidic systems for DNA analysis.

    PubMed

    Njoroge, Samuel K; Chen, Hui-Wen; Witek, Małgorzata A; Soper, Steven A

    2011-01-01

    The potential utility of genome-related research in terms of evolving basic discoveries in biology has generated widespread use of DNA diagnostics and DNA forensics and driven the accelerated development of fully integrated microfluidic systems for genome processing. To produce a microsystem with favorable performance characteristics for genetic-based analyses, several key operational elements must be strategically chosen, including device substrate material, temperature control, fluidic control, and reaction product readout. As a matter of definition, a microdevice is a chip that performs a single processing step, for example microchip electrophoresis. Several microdevices can be integrated to a single wafer, or combined on a control board as separate devices to form a microsystem. A microsystem is defined as a chip composed of at least two microdevices. Among the many documented analytical microdevices, those focused on the ability to perform the polymerase chain reaction (PCR) have been reported extensively due to the importance of this processing step in most genetic-based assays. Other microdevices that have been detailed in the literature include those for solid-phase extractions, microchip electrophoresis, and devices composed of DNA microarrays used for interrogating DNA primary structure. Great progress has also been made in the areas of chip fabrication, bonding and sealing to enclose fluidic networks, evaluation of different chip substrate materials, surface chemistries, and the architecture of reaction conduits for basic processing steps such as mixing. Other important elements that have been developed to realize functional systems include miniaturized readout formats comprising optical or electrochemical transduction and interconnect technologies. These discoveries have led to the development of fully autonomous and functional integrated systems for genome processing that can supply "sample in/answer out" capabilities. In this chapter, we focus on

  9. Multilayer-based lab-on-a-chip systems for perfused cell-based assays

    NASA Astrophysics Data System (ADS)

    Klotzbach, Udo; Sonntag, Frank; Grünzner, Stefan; Busek, Mathias; Schmieder, Florian; Franke, Volker

    2014-12-01

    A novel integrated technology chain of laser-microstructured multilayer foils for fast, flexible, and low-cost manufacturing of lab-on-a-chip devices especially for complex cell and tissue culture applications, which provides pulsatile fluid flow within physiological ranges at low media-to-cells ratio, was developed and established. Initially the microfluidic system is constructively divided into individual layers, which are formed by separate foils or plates. Based on the functional boundary conditions and the necessary properties of each layer, their corresponding foils and plates are chosen. In the third step, the foils and plates are laser microstructured and functionalized from both sides. In the fourth and last manufacturing step, the multiple plates and foils are joined using different bonding techniques like adhesive bonding, welding, etc. This multilayer technology together with pneumatically driven micropumps and valves permits the manufacturing of fluidic structures and perfusion systems, which spread out above multiple planes. Based on the established lab-on-a-chip platform for perfused cell-based assays, a multilayer microfluidic system with two parallel connected cell culture chambers was successfully implemented.

  10. The MainSTREAM component platform: a holistic approach to microfluidic system design.

    PubMed

    Sabourin, David; Skafte-Pedersen, Peder; Søe, Martin Jensen; Hemmingsen, Mette; Alberti, Massimo; Coman, Vasile; Petersen, Jesper; Emnéus, Jenny; Kutter, Jörg P; Snakenborg, Detlef; Jørgensen, Flemming; Clausen, Christian; Holmstrøm, Kim; Dufva, Martin

    2013-06-01

    A microfluidic component library for building systems driving parallel or serial microfluidic-based assays is presented. The components are a miniaturized eight-channel peristaltic pump, an eight-channel valve, sample-to-waste liquid management, and interconnections. The library of components was tested by constructing various systems supporting perfusion cell culture, automated DNA hybridizations, and in situ hybridizations. The results showed that the MainSTREAM components provided (1) a rapid, robust, and simple method to establish numerous fluidic inputs and outputs to various types of reaction chips; (2) highly parallel pumping and routing/valving capability; (3) methods to interface pumps and chip-to-liquid management systems; (4) means to construct a portable system; (5) reconfigurability/flexibility in system design; (6) means to interface to microscopes; and (7) compatibility with tested biological methods. It was found that LEGO Mindstorms motors, controllers, and software were robust, inexpensive, and an accessible choice as compared with corresponding custom-made actuators. MainSTREAM systems could operate continuously for weeks without leaks, contamination, or system failures. In conclusion, the MainSTREAM components described here meet many of the demands on components for constructing and using microfluidics systems. PMID:23015520

  11. Microfluidic System for Solution Array Based Bioassays

    SciTech Connect

    Dougherty, G M; Tok, J B; Pannu, S S; Rose, K A

    2006-02-10

    The objective of this project is to demonstrate new enabling technology for multiplex biodetection systems that are flexible, miniaturizable, highly automated, low cost, and high performance. It builds on prior successes at LLNL with particle-based solution arrays, such as those used in the Autonomous Pathogen Detection System (APDS) successfully field deployed to multiple locations nationwide. We report the development of a multiplex solution array immunoassay based upon engineered metallic nanorod particles. Nanobarcodes{reg_sign} particles are fabricated by sequential electrodeposition of dissimilar metals within porous alumina templates, yielding optically encoded striping patterns that can be read using standard laboratory microscope optics and PC-based image processing software. The addition of self-assembled monolayer (SAM) coatings and target-specific antibodies allows each encoded class of nanorod particles to be directed against a different antigen target. A prototype assay panel directed against bacterial, viral, and soluble protein targets demonstrates simultaneous detection at sensitivities comparable to state of the art immunoassays, with minimal cross-reactivity. Studies have been performed to characterize the colloidal properties (zeta potential) of the suspended nanorod particles as a function of pH, the ionic strength of the suspending solution, and surface functionalization state. Additional studies have produced means for the non-contact manipulation of the particles, including the insertion of magnetic nickel stripes within the encoding pattern, and control via externally applied electromagnetic fields. Using the results of these studies, the novel Nanobarcodes{reg_sign} based assay was implemented in a prototype automated system with the sample processing functions and optical readout performed on a microfluidic card. The unique physical properties of the nanorod particles enable the development of integrated microfluidic systems for

  12. Sample preparation system for microfluidic applications

    DOEpatents

    Mosier, Bruce P.; Crocker, Robert W.; Patel, Kamlesh D.; Harnett, Cindy K.

    2007-05-08

    An apparatus that couples automated injection with flow feedback to provide nanoliter accuracy in controlling microliter volumes. The apparatus comprises generally a source of hydraulic fluid pressure, a fluid isolator joined to the outlet of the hydraulic pressure source and a flow sensor to provide pressure-driven analyte metering. For operation generally and particularly in microfluidic systems the hydraulic pressure source is typically an electrokinetic (EK) pump that incorporates gasless electrodes. The apparatus is capable of metering sub-microliter volumes at flowrates of 1 100 .mu.L/min into microsystem load pressures of up to 1000 50 psi, respectively. Flowrates can be specified within 0.5 .mu.L/min and volumes as small as 80 nL can be metered.

  13. Single cell microfluidics for systems oncology

    NASA Astrophysics Data System (ADS)

    Fan, Rong

    2012-02-01

    The singular term ``cancer'' is never one kind of disease, but deceivingly encompasses a large number of heterogeneous disease states, which makes it impossible to completely treat cancer using a generic approach. Rather systems approaches are urgently required to assess cancer heterogeneity, stratify patients and enable the most effective, individualized treatment. The heterogeneity of tumors at the single cell level is reflected by the hierarchical complexity of the tumor microenvironment. To identify all the cellular components, including both tumor and infiltrating immune cells, and to delineate the associated cell-to-cell signaling network that dictates tumor initiation, progression and metastasis, we developed a single cell microfluidics chip that can analyze a panel of proteins that are potentially associated inter-cellular signaling network in tumor microenvironment from hundreds of single cells in parallel. This platform integrates two advanced technologies -- microfluidic single cell handling and ultra-high density protein array. This device was first tested for highly multiplexed profiling of secreted proteins including tumor-immune signaling molecules from monocytic leukemia cells. We observed profound cellular heterogeneity with all functional phenotypes quantitatively identified. Correlation analysis further indicated the existence of an intercellular cytokine network in which TNFα-induced secondary signaling cascades further increased functional cellular diversity. It was also exploited to evaluate polyfunctionality of tumor antigen-specific T cells from melanoma patients being treated with adoptive T cell transfer immunotherapy. This platform could be further extended to analyze both solid tumor cells (e.g. human lung carcinoma cells) and infiltrating immune cells (e.g. macrophages) so as to enable systems analysis of the complex tumor microenvironment from small amounts of clinical specimens, e.g. skinny needle biopsies. Thus, it could potentially

  14. Nifedipine and thallium-201 myocardial perfusion in progressive systemic sclerosis

    SciTech Connect

    Kahan, A.; Devaux, J.Y.; Amor, B.; Menkes, C.J.; Weber, S.; Nitenberg, A.; Venot, A.; Guerin, F.; Degeorges, M.; Roucayrol, J.C.

    1986-05-29

    Heart disease in patients with progressive systemic sclerosis may be due in part to myocardial ischemia caused by a disturbance of the coronary microcirculation. To determine whether abnormalities of myocardial perfusion in this disorder are potentially reversible, we evaluated the effect of the coronary vasodilator nifedipine on myocardial perfusion assessed by thallium-201 scanning in 20 patients. Thallium-201 single-photon-emission computerized tomography was performed under control conditions and 90 minutes after 20 mg of oral nifedipine. The mean (+/- SD) number of left ventricular segments with perfusion defects decreased from 5.3 +/- 2.0 to 3.3 +/- 2.2 after nifedipine (P = 0.0003). Perfusion abnormalities were quantified by a perfusion score (0 to 2.0) assigned to each left ventricular segment and by a global perfusion score (0 to 18) for the entire left ventricle. The mean perfusion score in segments with resting defects increased from 0.97 +/- 0.24 to 1.26 +/- 0.44 after nifedipine (P less than 0.00001). The mean global perfusion score increased from 11.2 +/- 1.7 to 12.8 +/- 2.4 after nifedipine (P = 0.003). The global perfusion score increased by at least 2.0 in 10 patients and decreased by at least 2.0 in only 1. These observations reveal short-term improvement in thallium-201 myocardial perfusion with nifedipine in patients with progressive systemic sclerosis. The results are consistent with a potentially reversible abnormality of coronary vasomotion in this disorder, but the long-term therapeutic effects of nifedipine remain to be determined.

  15. A hybrid MEMS-based microfluidic system for cancer diagnosis

    NASA Astrophysics Data System (ADS)

    Ortiz, Pedro; Keegan, Neil; Spoors, Julia; Hedley, John; Harris, Alun; Burdess, Jim; Burnett, Richard; Velten, Thomas; Biehl, Margit; Knoll, Thorsten; Haberer, Werner; Solomon, Matthew; Campitelli, Andrew; McNeil, Calum

    2008-12-01

    A microfluidic system for cancer diagnosis based around a core MEMS biosensor technology is presented in this paper. The principle of the MEMS biosensor is introduced and the functionalisation strategy for cancer marker recognition is described. In addition, the successful packaging and integration of functional MEMS biosensor devices are reported herein. This ongoing work represents one of the first hybrid systems to integrate a PCB packaged silicon MEMS device into a disposable microfluidic cartridge.

  16. Tailoring microfluidic systems for organ-like cell culture applications using multiphysics simulations

    NASA Astrophysics Data System (ADS)

    Hagmeyer, Britta; Schütte, Julia; Böttger, Jan; Gebhardt, Rolf; Stelzle, Martin

    2013-03-01

    Replacing animal testing with in vitro cocultures of human cells is a long-term goal in pre-clinical drug tests used to gain reliable insight into drug-induced cell toxicity. However, current state-of-the-art 2D or 3D cell cultures aiming at mimicking human organs in vitro still lack organ-like morphology and perfusion and thus organ-like functions. To this end, microfluidic systems enable construction of cell culture devices which can be designed to more closely resemble the smallest functional unit of organs. Multiphysics simulations represent a powerful tool to study the various relevant physical phenomena and their impact on functionality inside microfluidic structures. This is particularly useful as it allows for assessment of system functions already during the design stage prior to actual chip fabrication. In the HepaChip®, dielectrophoretic forces are used to assemble human hepatocytes and human endothelial cells in liver sinusoid-like structures. Numerical simulations of flow distribution, shear stress, electrical fields and heat dissipation inside the cell assembly chambers as well as surface wetting and surface tension effects during filling of the microchannel network supported the design of this human-liver-on-chip microfluidic system for cell culture applications. Based on the device design resulting thereof, a prototype chip was injection-moulded in COP (cyclic olefin polymer). Functional hepatocyte and endothelial cell cocultures were established inside the HepaChip® showing excellent metabolic and secretory performance.

  17. Integrated Electroosmotic Perfusion of Tissue with Online Microfluidic Analysis to Track the Metabolism of Cystamine, Pantethine and Coenzyme A

    PubMed Central

    Wu, Juanfang; Sandberg, Mats; Weber, Stephen G.

    2014-01-01

    We have developed an approach that integrates electroosmotic perfusion of tissue with a substrate-containing solution and online microfluidic analysis of products, in this case thiols. Using this approach we have tracked the metabolism of cystamine, pantethine and CoA in the extracellular space of organotypic hippocampal slice cultures (OHSCs). Currently, little is known about coenzyme A (CoA) biodegradation and even less is known about the regulation and kinetic characteristics for this sequential multi-enzyme reaction. We found that the steady state percentage yields of cysteamine from cystamine and pantethine during the transit through OHSCs were 91% ± 4% (SEM) and 0.01%–0.03%, respectively. The large difference in the yields of cysteamine can be used to explain the drugs’ different toxicities and clinical effectiveness against cystinosis. The kinetic parameters of the enzyme reaction catalyzed by the ectoenzyme pantetheinase are KM,C/α = 4.4 ± 1.1 mM and Vmax,C = 29 ± 3 nM/s, where α is the percentage yield of pantethine to pantetheine through disulfide exchange. We estimate that the percentage yield of pantethine to pantetheine through disulfide exchange is approximately 0.5%. Based on the formation rate of cysteamine in the OHSCs, we obtained the overall apparent Michaelis constant and maximum reaction rate for sequential, extracellular CoA degradation in an in situ environment, which are K′M = 16 ± 4 μM, V′max = 7.1 ± 0.5 nM/s. Kinetic parameters obtained in situ, although difficult to measure, are better representations of the biochemical flux in the living organism than those from isolated enzymes in vitro. PMID:24215585

  18. Integrated electroosmotic perfusion of tissue with online microfluidic analysis to track the metabolism of cystamine, pantethine, and coenzyme A.

    PubMed

    Wu, Juanfang; Sandberg, Mats; Weber, Stephen G

    2013-12-17

    We have developed an approach that integrates electroosmotic perfusion of tissue with a substrate-containing solution and online microfluidic analysis of products, in this case thiols. Using this approach we have tracked the metabolism of cystamine, pantethine and CoA in the extracellular space of organotypic hippocampal slice cultures (OHSCs). Currently, little is known about coenzyme A (CoA) biodegradation and even less is known about the regulation and kinetic characteristics for this sequential multienzyme reaction. We found that the steady state percentage yields of cysteamine from cystamine and pantethine during the transit through OHSCs were 91% ± 4% (SEM) and 0.01%-0.03%, respectively. The large difference in the yields of cysteamine can be used to explain the drugs' different toxicities and clinical effectiveness against cystinosis. The kinetic parameters of the enzyme reaction catalyzed by the ectoenzyme pantetheinase are KM,C/α = 4.4 ± 1.1 mM and Vmax,C = 29 ± 3 nM/s, where α is the percentage yield of pantethine to pantetheine through disulfide exchange. We estimate that the percentage yield of pantethine to pantetheine through disulfide exchange is approximately 0.5%. Based on the formation rate of cysteamine in the OHSCs, we obtained the overall apparent Michaelis constant and maximum reaction rate for sequential, extracellular CoA degradation in an in situ environment, which are K'M = 16 ± 4 μM, V'max = 7.1 ± 0.5 nM/s. Kinetic parameters obtained in situ, although difficult to measure, are better representations of the biochemical flux in the living organism than those from isolated enzymes in vitro. PMID:24215585

  19. Chemically generated convective transport in microfluidic system

    NASA Astrophysics Data System (ADS)

    Shklyaev, Oleg; Das, Sambeeta; Altemose, Alicia; Shum, Henry; Balazs, Anna; Sen, Ayusman

    High precision manipulation of small volumes of fluid, containing suspended micron sized objects like cells, viruses, and large molecules, is one of the main goals in designing modern lab-on-a-chip devices which can find a variety of chemical and biological applications. To transport the cargo toward sensing elements, typical microfluidic devices often use pressure driven flows. Here, we propose to use enzymatic chemical reactions which decompose reagent into less dense products and generate flows that can transport particles. Density variations that lead to flow in the assigned direction are created between the place where reagent is fed into the solution and the location where it is decomposed by enzymes attached to the surface of the microchannel. When the reagent is depleted, the fluid motion stops and particles sediment to the bottom. We demonstrate how the choice of chemicals, leading to specific reaction rates, can affect the transport properties. In particular, we show that the intensity of the fluid flow, the final location of cargo, and the time for cargo delivery are controlled by the amount and type of reagent in the system.

  20. Acoustothermal heating of polydimethylsiloxane microfluidic system.

    PubMed

    Ha, Byung Hang; Lee, Kang Soo; Destgeer, Ghulam; Park, Jinsoo; Choung, Jin Seung; Jung, Jin Ho; Shin, Jennifer Hyunjong; Sung, Hyung Jin

    2015-01-01

    We report an observation of rapid (exceeding 2,000 K/s) heating of polydimethylsiloxane (PDMS), one of the most popular microchannel materials, under cyclic loadings at high (~MHz) frequencies. A microheater was developed based on the finding. The heating mechanism utilized vibration damping in PDMS induced by sound waves that were generated and precisely controlled using a conventional surface acoustic wave (SAW) microfluidic system. The refraction of SAW into the PDMS microchip, called the leaky SAW, takes a form of bulk wave and rapidly heats the microchannels in a volumetric manner. The penetration depths were measured to range from 210 μm to 1290 μm, enough to cover most sizes of microchannels. The energy conversion efficiency was SAW frequency-dependent and measured to be the highest at around 30 MHz. Independent actuation of each interdigital transducer (IDT) enabled independent manipulation of SAWs, permitting spatiotemporal control of temperature on the microchip. All the advantages of this microheater facilitated a two-step continuous flow polymerase chain reaction (CFPCR) to achieve the billion-fold amplification of a 134 bp DNA amplicon in less than 3 min. PMID:26138310

  1. Acoustothermal heating of polydimethylsiloxane microfluidic system

    NASA Astrophysics Data System (ADS)

    Ha, Byung Hang; Lee, Kang Soo; Destgeer, Ghulam; Park, Jinsoo; Choung, Jin Seung; Jung, Jin Ho; Shin, Jennifer Hyunjong; Sung, Hyung Jin

    2015-07-01

    We report an observation of rapid (exceeding 2,000 K/s) heating of polydimethylsiloxane (PDMS), one of the most popular microchannel materials, under cyclic loadings at high (~MHz) frequencies. A microheater was developed based on the finding. The heating mechanism utilized vibration damping in PDMS induced by sound waves that were generated and precisely controlled using a conventional surface acoustic wave (SAW) microfluidic system. The refraction of SAW into the PDMS microchip, called the leaky SAW, takes a form of bulk wave and rapidly heats the microchannels in a volumetric manner. The penetration depths were measured to range from 210 μm to 1290 μm, enough to cover most sizes of microchannels. The energy conversion efficiency was SAW frequency-dependent and measured to be the highest at around 30 MHz. Independent actuation of each interdigital transducer (IDT) enabled independent manipulation of SAWs, permitting spatiotemporal control of temperature on the microchip. All the advantages of this microheater facilitated a two-step continuous flow polymerase chain reaction (CFPCR) to achieve the billion-fold amplification of a 134 bp DNA amplicon in less than 3 min.

  2. Acoustothermal heating of polydimethylsiloxane microfluidic system

    PubMed Central

    Ha, Byung Hang; Lee, Kang Soo; Destgeer, Ghulam; Park, Jinsoo; Choung, Jin Seung; Jung, Jin Ho; Shin, Jennifer Hyunjong; Sung, Hyung Jin

    2015-01-01

    We report an observation of rapid (exceeding 2,000 K/s) heating of polydimethylsiloxane (PDMS), one of the most popular microchannel materials, under cyclic loadings at high (~MHz) frequencies. A microheater was developed based on the finding. The heating mechanism utilized vibration damping in PDMS induced by sound waves that were generated and precisely controlled using a conventional surface acoustic wave (SAW) microfluidic system. The refraction of SAW into the PDMS microchip, called the leaky SAW, takes a form of bulk wave and rapidly heats the microchannels in a volumetric manner. The penetration depths were measured to range from 210 μm to 1290 μm, enough to cover most sizes of microchannels. The energy conversion efficiency was SAW frequency-dependent and measured to be the highest at around 30 MHz. Independent actuation of each interdigital transducer (IDT) enabled independent manipulation of SAWs, permitting spatiotemporal control of temperature on the microchip. All the advantages of this microheater facilitated a two-step continuous flow polymerase chain reaction (CFPCR) to achieve the billion-fold amplification of a 134 bp DNA amplicon in less than 3 min. PMID:26138310

  3. Active liquid degassing in microfluidic systems.

    PubMed

    Karlsson, J Mikael; Gazin, Muriel; Laakso, Sanna; Haraldsson, Tommy; Malhotra-Kumar, Surbhi; Mäki, Minna; Goossens, Herman; van der Wijngaart, Wouter

    2013-11-21

    We present a method for efficient air bubble removal in microfluidic applications. Air bubbles are extracted from a liquid chamber into a vacuum chamber through a semipermeable membrane, consisting of PDMS coated with amorphous Teflon(®) AF 1600. Whereas air is efficiently extracted through the membrane, water loss is greatly reduced by the Teflon even at elevated temperatures. We present the water loss and permeability change with the amount of added Teflon AF to the membrane. Also, we demonstrate bubble-free, multiplex DNA amplification using PCR in a PDMS microfluidic device. PMID:24056885

  4. Microfluidic systems for stem cell-based neural tissue engineering.

    PubMed

    Karimi, Mahdi; Bahrami, Sajad; Mirshekari, Hamed; Basri, Seyed Masoud Moosavi; Nik, Amirala Bakhshian; Aref, Amir R; Akbari, Mohsen; Hamblin, Michael R

    2016-07-01

    Neural tissue engineering aims at developing novel approaches for the treatment of diseases of the nervous system, by providing a permissive environment for the growth and differentiation of neural cells. Three-dimensional (3D) cell culture systems provide a closer biomimetic environment, and promote better cell differentiation and improved cell function, than could be achieved by conventional two-dimensional (2D) culture systems. With the recent advances in the discovery and introduction of different types of stem cells for tissue engineering, microfluidic platforms have provided an improved microenvironment for the 3D-culture of stem cells. Microfluidic systems can provide more precise control over the spatiotemporal distribution of chemical and physical cues at the cellular level compared to traditional systems. Various microsystems have been designed and fabricated for the purpose of neural tissue engineering. Enhanced neural migration and differentiation, and monitoring of these processes, as well as understanding the behavior of stem cells and their microenvironment have been obtained through application of different microfluidic-based stem cell culture and tissue engineering techniques. As the technology advances it may be possible to construct a "brain-on-a-chip". In this review, we describe the basics of stem cells and tissue engineering as well as microfluidics-based tissue engineering approaches. We review recent testing of various microfluidic approaches for stem cell-based neural tissue engineering. PMID:27296463

  5. Optical two-beam traps in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Berg-Sørensen, Kirstine

    2016-08-01

    An attractive solution for optical trapping and stretching by means of two counterpropagating laser beams is to embed waveguides or optical fibers in a microfluidic system. The microfluidic system can be constructed in different materials, ranging from soft polymers that may easily be cast in a rapid prototyping manner, to hard polymers that could even be produced by injection moulding, or to silica in which waveguides may either be written directly, or with grooves for optical fibers. Here, we review different solutions to the system and also show results obtained in a polymer chip with DUV written waveguides and in an injection molded polymer chip with grooves for optical fibers.

  6. Microfluidics-based integrated airborne pathogen detection systems

    NASA Astrophysics Data System (ADS)

    Northrup, M. Allen; Alleman-Sposito, Jennifer; Austin, Todd; Devitt, Amy; Fong, Donna; Lin, Phil; Nakao, Brian; Pourahmadi, Farzad; Vinas, Mary; Yuan, Bob

    2006-09-01

    Microfluidic Systems is focused on building microfluidic platforms that interface front-end mesofluidics to handle real world sample volumes for optimal sensitivity coupled to microfluidic circuitry to process small liquid volumes for complex reagent metering, mixing, and biochemical analysis, particularly for pathogens. MFSI is the prime contractor on two programs for the US Department of Homeland Security: BAND (Bioagent Autonomous Networked Detector) and IBADS (Instantaneous Bio-Aerosol Detection System). The goal of BAND is to develop an autonomous system for monitoring the air for known biological agents. This consists of air collection, sample lysis, sample purification, detection of DNA, RNA, and toxins, and a networked interface to report the results. For IBADS, MFSI is developing the confirmatory device which must verify the presence of a pathogen with 5 minutes of an air collector/trigger sounding an alarm. Instrument designs and biological assay results from both BAND and IBADS will be presented.

  7. Microfluidic systems for electrochemical and biological studies

    SciTech Connect

    Ackler, H., LLNL

    1998-05-01

    Microfluidic devices with microelectrodes have the potential to enable studies of phenomena at size scales where behavior may be dominated by different mechanisms than at macroscales. Through our work developing microfluidic devices for dielectrophoretic separation and sensing of cells and particles, we have fabricated devices from which general or more specialized research devices may be derived. Fluid channels from 80 {micro}m wide X 20 {micro}m deep to 1 mm wide to 200 {micro}m deep have been fabricated in glass, with lithographically patterned electrodes from 10 to 80 {micro}m wide on one or both sides on the channels and over topographies tens of microns in heights. the devices are designed to easily interface to electronic and fluidic interconnect packages that permit reuse of devices, rather than one-time use, crude glue-based methods. Such devices may be useful for many applications of interest to the electrochemical and biological community.

  8. A metering rotary nanopump for microfluidic systems

    PubMed Central

    Darby, Scott G.; Moore, Matthew R.; Friedlander, Troy A.; Schaffer, David K.; Reiserer, Ron S.; Wikswo, John P.

    2014-01-01

    We describe the design, fabrication, and testing of a microfabricated metering rotary nanopump for the purpose of driving fluid flow in microfluidic devices. The miniature peristaltic pump is composed of a set of microfluidic channels wrapped in a helix around a central cam shaft in which a non-cylindrical cam rotates. The cam compresses the helical channels to induce peristaltic flow as it is rotated. The polydimethylsiloxane (PDMS) nanopump design is able to produce intermittent delivery or removal of several nanoliters of fluid per revolution as well as consistent continuous flow rates ranging from as low as 15 nL/min to above 1.0 µL/min. At back pressures encountered in typical microfluidic devices, the pump acts as a high impedance flow source. The durability, biocompatibility, ease of integration with soft-lithographic fabrication, the use of a simple rotary motor instead of multiple synchronized pneumatic or mechanical actuators, and the absence of power consumption or fluidic conductance in the resting state all contribute to a compact pump with a low cost of fabrication and versatile implementation. This suggests that the pump design may be useful for a wide variety of biological experiments and point of care devices. PMID:20959938

  9. A metering rotary nanopump for microfluidic systems.

    PubMed

    Darby, Scott G; Moore, Matthew R; Friedlander, Troy A; Schaffer, David K; Reiserer, Ron S; Wikswo, John P; Seale, Kevin T

    2010-12-01

    We describe the design, fabrication, and testing of a microfabricated metering rotary nanopump for the purpose of driving fluid flow in microfluidic devices. The miniature peristaltic pump is composed of a set of microfluidic channels wrapped in a helix around a central camshaft in which a non-cylindrical cam rotates. The cam compresses the helical channels to induce peristaltic flow as it is rotated. The polydimethylsiloxane (PDMS) nanopump design is able to produce intermittent delivery or removal of several nanolitres of fluid per revolution as well as consistent continuous flow rates ranging from as low as 15 nL min(-1) to above 1.0 µL min(-1). At back pressures encountered in typical microfluidic devices, the pump acts as a high impedance flow source. The durability, biocompatibility, ease of integration with soft-lithographic fabrication, the use of a simple rotary motor instead of multiple synchronized pneumatic or mechanical actuators, and the absence of power consumption or fluidic conductance in the resting state all contribute to a compact pump with a low cost of fabrication and versatile implementation. This suggests that the pump design may be useful for a wide variety of biological experiments and point of care devices. PMID:20959938

  10. A Microfluidics-HPLC/Differential Mobility Spectrometer Macromolecular Detection System for Human and Robotic Missions

    NASA Technical Reports Server (NTRS)

    Coy, S. L.; Killeen, K.; Han, J.; Eiceman, G. A.; Kanik, I.; Kidd, R. D.

    2011-01-01

    Our goal is to develop a unique, miniaturized, solute analyzer based on microfluidics technology. The analyzer consists of an integrated microfluidics High Performance Liquid Chromatographic chip / Differential Mobility Spectrometer (?HPLCchip/ DMS) detection system

  11. Micro Electromechanical Systems (MEMS) Based Microfluidic Devices for Biomedical Applications

    PubMed Central

    Ashraf, Muhammad Waseem; Tayyaba, Shahzadi; Afzulpurkar, Nitin

    2011-01-01

    Micro Electromechanical Systems (MEMS) based microfluidic devices have gained popularity in biomedicine field over the last few years. In this paper, a comprehensive overview of microfluidic devices such as micropumps and microneedles has been presented for biomedical applications. The aim of this paper is to present the major features and issues related to micropumps and microneedles, e.g., working principles, actuation methods, fabrication techniques, construction, performance parameters, failure analysis, testing, safety issues, applications, commercialization issues and future prospects. Based on the actuation mechanisms, the micropumps are classified into two main types, i.e., mechanical and non-mechanical micropumps. Microneedles can be categorized according to their structure, fabrication process, material, overall shape, tip shape, size, array density and application. The presented literature review on micropumps and microneedles will provide comprehensive information for researchers working on design and development of microfluidic devices for biomedical applications. PMID:21747700

  12. System-level simulation of liquid filling in microfluidic chips.

    PubMed

    Song, Hongjun; Wang, Yi; Pant, Kapil

    2011-06-01

    Liquid filling in microfluidic channels is a complex process that depends on a variety of geometric, operating, and material parameters such as microchannel geometry, flow velocity∕pressure, liquid surface tension, and contact angle of channel surface. Accurate analysis of the filling process can provide key insights into the filling time, air bubble trapping, and dead zone formation, and help evaluate trade-offs among the various design parameters and lead to optimal chip design. However, efficient modeling of liquid filling in complex microfluidic networks continues to be a significant challenge. High-fidelity computational methods, such as the volume of fluid method, are prohibitively expensive from a computational standpoint. Analytical models, on the other hand, are primarily applicable to idealized geometries and, hence, are unable to accurately capture chip level behavior of complex microfluidic systems. This paper presents a parametrized dynamic model for the system-level analysis of liquid filling in three-dimensional (3D) microfluidic networks. In our approach, a complex microfluidic network is deconstructed into a set of commonly used components, such as reservoirs, microchannels, and junctions. The components are then assembled according to their spatial layout and operating rationale to achieve a rapid system-level model. A dynamic model based on the transient momentum equation is developed to track the liquid front in the microchannels. The principle of mass conservation at the junction is used to link the fluidic parameters in the microchannels emanating from the junction. Assembly of these component models yields a set of differential and algebraic equations, which upon integration provides temporal information of the liquid filling process, particularly liquid front propagation (i.e., the arrival time). The models are used to simulate the transient liquid filling process in a variety of microfluidic constructs and in a multiplexer, representing a

  13. Perfusion Electronic Record Documentation Using Epic Systems Software

    PubMed Central

    Steffens, Thomas G.; Gunser, John M.; Saviello, George M.

    2015-01-01

    Abstract: This paper describes the design and use of Epic Systems software for documentation of perfusion activities as part of the patient electronic medical record. The University of Wisconsin Hospital and Clinics adapted the Anesthesia software module and developed an integrated perfusion/anesthesia record for the documentation of cardiac and non-cardiac surgical procedures. This project involved multiple committees, approvals, and training to successfully implement. This article will describe our documentation options, concepts, design, challenges, training, and implementation during our initial experience. PMID:26834288

  14. Nanostructured microfluidic digestion system for rapid high-performance proteolysis

    PubMed Central

    Cheng, Gong; Hao, Si-Jie; Yu, Xu

    2014-01-01

    A novel microfluidic protein digestion system with nanostructured and bioactive inner surface was constructed by an easy biomimetic self-assembly strategy for rapid and effective proteolysis in 2 minutes, which is faster than the conventional overnight digestion methods. It is expected that this work would contribute to rapid online digestion in future high-throughput proteomics. PMID:25511010

  15. Cusps and spouts in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Duboin, Aurélien; Malloggi, Florent; Monti, Fabrice; Tabeling, Patrick

    2011-11-01

    By injecting mineral oil (inner phase) and polymer solutions (outer phase), in a microfluidic flow focusing geometry, we observed the formation of cusps. These cusps undergo a transition from a steady state, to a thin cylindrical spout (oil in polymer). These oil spouts, do not touch the walls, and are surprisingly stable (they do not break into droplets). We study the nature of the cusp-spout transition, and find it is of first order. By taking advantage of the stability of the jet, we expect to synthesize micro-wires with this approach.

  16. Characterizing potential heart agents with an isolated perfused heart system

    SciTech Connect

    Pendleton, D.B.; Sands, H.; Gallagher, B.M.; Camin, L.L.

    1984-01-01

    The authors have used an isolated perfused heart system for characterizing potential myocardial perfusion radiopharamaceuticals. Rabbit or guinea pig (GP) hearts are removed and perfused through the aorta with a blood-free buffer. Heart rate and ventricular pressure are monitored as indices of viability. Tc-99m-MAA is 96-100% retained in these hearts, and Tc-99m human serum albumin shows less than 5% extraction. Tl-201 is 30-40% extracted. It is known that in-vivo, Tc-99m(dmpe)/sub 2/Cl/sub 2//sup +/ is taken up by rabbit heart but not by GP or human heart. Analogous results are obtained with the isolated perfused heart model, where the complex is extracted well by the isolated rabbit heart (24%) but not by the GP heart (<5%). Values are unchanged if human, rabbit or GP blood is mixed and co-injected with the complex. Tc-99m)dmpe)/sub 3//sup +/ is also taken up by rabbit but not by GP hearts in-vivo. However, isolated perfused hearts of both species extract this complex well (45-52%). Heart uptake is diminished to <7% if the complex is pre-equilibrated with human blood. GP blood produces a moderate inhibition (in GP hearts only) and rabbit blood has no effect. This suggests that a human or GP blood factor may have a significant effect on heart uptake of this complex. Tc-99m(CN-t-butyl)/sub 6//sup +/ is taken up well by both rabbit and GP hearts in-vivo, and is extracted 100% by both isolated perfused hearts. Heart retention remains high (73-75%) in the presence of human blood.

  17. Microfluidics-Based Laser Guided Cell-Micropatterning System

    PubMed Central

    Erdman, Nick; Schmidt, Lucas; Qin, Wan; Yang, Xiaoqi; Lin, Yongliang; DeSilva, Mauris N; Gao, Bruce Z.

    2014-01-01

    The ability to place individual cells into an engineered microenvironment in a cell-culture model is critical for the study of in vivo-relevant cell-cell and cell-extracellular matrix interactions. Microfluidics provides a high-throughput modality to inject various cell types into a microenvironment. Laser guided systems provide the high spatial and temporal resolution necessary for single-cell micropatterning. Combining these two techniques, the authors designed, constructed, tested, and evaluated 1) a novel removable microfluidics-based cell-delivery biochip and 2) a combined system that uses the novel biochip coupled with a laser guided cell-micropatterning system to place individual cells into both 2D and 3D arrays. Cell-suspensions of chick forebrain neurons and glial cells were loaded into their respective inlet reservoirs and traversed the microfluidic channels until reaching the outlet ports. Individual cells were trapped and guided from the outlet of a microfluidic channel to a target site on the cell-culture substrate. At the target site, 2D and 3D pattern arrays were constructed with micron-level accuracy. Single-cell manipulation was accomplished at a rate of 150 μm/s in the radial plane and 50 μm/s in the axial direction of the laser beam. Results demonstrated that a single-cell can typically be patterned in 20-30 seconds, and that highly accurate and reproducible cellular arrays and systems can be achieved through coupling the microfluidics-based cell-delivery biochip with the laser guided system. PMID:25190714

  18. Microfluidics-based laser cell-micropatterning system.

    PubMed

    Erdman, Nick; Schmidt, Lucas; Qin, Wan; Yang, Xiaoqi; Lin, Yongliang; DeSilva, Mauris N; Gao, Bruce Z

    2014-09-01

    The ability to place individual cells into an engineered microenvironment in a cell-culture model is critical for the study of in vivo relevant cell-cell and cell-extracellular matrix interactions. Microfluidics provides a high-throughput modality to inject various cell types into a microenvironment. Laser guided systems provide the high spatial and temporal resolution necessary for single-cell micropatterning. Combining these two techniques, the authors designed, constructed, tested and evaluated (1) a novel removable microfluidics-based cell-delivery biochip and (2) a combined system that uses the novel biochip coupled with a laser guided cell-micropatterning system to place individual cells into both two-dimensional (2D) and three-dimensional (3D) arrays. Cell-suspensions of chick forebrain neurons and glial cells were loaded into their respective inlet reservoirs and traversed the microfluidic channels until reaching the outlet ports. Individual cells were trapped and guided from the outlet of a microfluidic channel to a target site on the cell-culture substrate. At the target site, 2D and 3D pattern arrays were constructed with micron-level accuracy. Single-cell manipulation was accomplished at a rate of 150 μm s(-1) in the radial plane and 50 μm s(-1) in the axial direction of the laser beam. Results demonstrated that a single-cell can typically be patterned in 20-30 s, and that highly accurate and reproducible cellular arrays and systems can be achieved through coupling the microfluidics-based cell-delivery biochip with the laser guided system. PMID:25190714

  19. Microfluidic Radiometal Labeling Systems for Biomolecules

    SciTech Connect

    Reichert, D E; Kenis, P J. A.

    2011-12-29

    In a typical labeling procedure with radiometals, such as Cu-64 and Ga-68; a very large (~ 100-fold) excess of the non-radioactive reactant (precursor) is used to promote rapid and efficient incorporation of the radioisotope into the PET imaging agent. In order to achieve high specific activities, careful control of reaction conditions and extensive chromatographic purifications are required in order to separate the labeled compounds from the cold precursors. Here we propose a microfluidic approach to overcome these problems, and achieve high specific activities in a more convenient, semi-automated fashion and faster time frame. Microfluidic reactors, consisting of a network of micron-sized channels (typical dimensions in the range 10 - 300¼m), filters, separation columns, electrodes and reaction loops/chambers etched onto a solid substrate, are now emerging as an extremely useful technology for the intensification and miniaturization of chemical processes. The ability to manipulate, process and analyze reagent concentrations and reaction interfaces in both space and time within the channel network of a microreactor provides the fine level of reaction control that is desirable in PET radiochemistry practice. These factors can bring radiometal labeling, specifically the preparation of radio-labeled biomolecules such as antibodies, much closer to their theoretical maximum specific activities.

  20. 21 CFR 876.5880 - Isolated kidney perfusion and transport system and accessories.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Isolated kidney perfusion and transport system and....5880 Isolated kidney perfusion and transport system and accessories. (a) Identification. An isolated kidney perfusion and transport system and accessories is a device that is used to support a donated or...

  1. 21 CFR 876.5880 - Isolated kidney perfusion and transport system and accessories.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Isolated kidney perfusion and transport system and....5880 Isolated kidney perfusion and transport system and accessories. (a) Identification. An isolated kidney perfusion and transport system and accesssories is a device that is used to support a donated or...

  2. 21 CFR 876.5880 - Isolated kidney perfusion and transport system and accessories.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Isolated kidney perfusion and transport system and....5880 Isolated kidney perfusion and transport system and accessories. (a) Identification. An isolated kidney perfusion and transport system and accessories is a device that is used to support a donated or...

  3. 21 CFR 876.5880 - Isolated kidney perfusion and transport system and accessories.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Isolated kidney perfusion and transport system and....5880 Isolated kidney perfusion and transport system and accessories. (a) Identification. An isolated kidney perfusion and transport system and accesssories is a device that is used to support a donated or...

  4. 21 CFR 876.5880 - Isolated kidney perfusion and transport system and accessories.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Isolated kidney perfusion and transport system and....5880 Isolated kidney perfusion and transport system and accessories. (a) Identification. An isolated kidney perfusion and transport system and accesssories is a device that is used to support a donated or...

  5. Myocardial perfusion abnormalities in asymptomatic patients with systemic lupus erythematosus

    SciTech Connect

    Hosenpud, J.D.; Montanaro, A.; Hart, M.V.; Haines, J.E.; Specht, H.D.; Bennett, R.M.; Kloster, F.E.

    1984-08-01

    Accelerated coronary artery disease and myocardial infarction in young patients with systemic lupus erythematosus is well documented; however, the prevalence of coronary involvement is unknown. Accordingly, 26 patients with systemic lupus were selected irrespective of previous cardiac history to undergo exercise thallium-201 cardiac scintigraphy. Segmental perfusion abnormalities were present in 10 of the 26 studies (38.5 percent). Five patients had reversible defects suggesting ischemia, four patients had persistent defects consistent with scar, and one patient had both reversible and persistent defects in two areas. There was no correlation between positive thallium results and duration of disease, amount of corticosteroid treatment, major organ system involvement or age. Only a history of pericarditis appeared to be associated with positive thallium-201 results (p less than 0.05). It is concluded that segmental myocardial perfusion abnormalities are common in patients with systemic lupus erythematosus. Whether this reflects large-vessel coronary disease or small-vessel abnormalities remains to be determined.

  6. Artificial Lymphatic Drainage Systems for Vascularized Microfluidic Scaffolds

    PubMed Central

    Wong, Keith H. K.; Truslow, James G.; Khankhel, Aimal H.; Chan, Kelvin L. S.; Tien, Joe

    2012-01-01

    The formation of a stably perfused microvasculature continues to be a major challenge in tissue engineering. Previous work has suggested the importance of a sufficiently large transmural pressure in maintaining vascular stability and perfusion. Here we show that a system of empty channels that provides a drainage function analogous to that of lymphatic microvasculature in vivo can stabilize vascular adhesion and maintain perfusion rate in dense, hydraulically resistive fibrin scaffolds in vitro. In the absence of drainage, endothelial delamination increased as scaffold density increased from 6 mg/mL to 30 mg/mL and scaffold hydraulic conductivity decreased by a factor of twenty. Single drainage channels exerted only localized vascular stabilization, the extent of which depended on the distance between vessel and drainage as well as scaffold density. Computational modeling of these experiments yielded an estimate of 0.40–1.36 cm H2O for the minimum transmural pressure required for vascular stability. We further designed and constructed fibrin patches (0.8 by 0.9 cm2) that were perfused by a parallel array of vessels and drained by an orthogonal array of drainage channels; only with the drainage did the vessels display long-term stability and perfusion. This work underscores the importance of drainage in vascularization, especially when a dense, hydraulically resistive scaffold is used. PMID:23281125

  7. A microfluidic system for dynamic yeast cell imaging.

    PubMed

    Lee, Philip J; Helman, Noah C; Lim, Wendell A; Hung, Paul J

    2008-01-01

    The investigation of cellular processes and gene regulatory networks within living cells requires the development of improved technology for dynamic, single cell imaging. Here, we demonstrate a microfluidic system capable of mechanical trapping of yeast cells with continuous flow and flow switching capability during time-lapse high magnification fluorescence imaging. The novel functionality of the system was validated by observing the response of pheromone-induced expression of GFP in Saccharomyces cerevisiae. PMID:18254385

  8. Imaging label-free biosensor with microfluidic system

    NASA Astrophysics Data System (ADS)

    Jahns, S.; Glorius, P.; Hansen, M.; Nazirizadeh, Y.; Gerken, M.

    2015-06-01

    We present a microfluidic system suitable for parallel label-free detection of several biomarkers utilizing a compact imaging measurement system. The microfluidic system contains a filter unit to separate the plasma from human blood and a functionalized, photonic crystal slab sensor chip. The nanostructure of the photonic crystal slab sensor chip is fabricated by nanoimprint lithography of a period grating surface into a photoresist and subsequent deposition of a TiO2 layer. Photonic crystal slabs are slab waveguides supporting quasi-guided modes coupling to far-field radiation, which are sensitive to refractive index changes due to biomarker binding on the functionalized surface. In our imaging read-out system the resulting resonance shift of the quasi-guided mode in the transmission spectrum is converted into an intensity change detectable with a simple camera. By continuously taking photographs of the sensor surface local intensity changes are observed revealing the binding kinetics of the biomarker to its specific target. Data from two distinct measurement fields are used for evaluation. For testing the sensor chip, 1 μM biotin as well as 1 μM recombinant human CD40 ligand were immobilized in spotsvia amin coupling to the sensor surface. Each binding experiment was performed with 250 nM streptavidin and 90 nM CD40 ligand antibody dissolved in phosphate buffered saline. In the next test series, a functionalized sensor chip was bonded onto a 15 mm x 15 mm opening of the 75 mm x 25 mm x 2 mm microfluidic system. We demonstrate the functionality of the microfluidic system for filtering human blood such that only blood plasma was transported to the sensor chip. The results of first binding experiments in buffer with this test chip will be presented.

  9. Centrifugal Microfluidic System for Nucleic Acid Amplification and Detection

    PubMed Central

    Miao, Baogang; Peng, Niancai; Li, Lei; Li, Zheng; Hu, Fei; Zhang, Zengming; Wang, Chaohui

    2015-01-01

    We report here the development of a rapid PCR microfluidic system comprising a double-shaft turntable and centrifugal-based disc that rapidly drives the PCR mixture between chambers set at different temperatures, and the bidirectional flow improved the space utilization of the disc. Three heating resistors and thermistors maintained uniform, specific temperatures for the denaturation, annealing, and extension steps of the PCR. Infrared imaging showed that there was little thermal interference between reaction chambers; the system enabled the cycle number and reaction time of each step to be independently adjusted. To validate the function and efficiency of the centrifugal microfluidic system, a 350-base pair target gene from the hepatitis B virus was amplified and quantitated by fluorescence detection. By optimizing the cycling parameters, the reaction time was reduced to 32 min as compared to 120 min for a commercial PCR machine. DNA samples with concentrations ranging from 10 to 106 copies/mL could be quantitatively analyzed using this system. This centrifugal-based microfluidic platform is a useful system and possesses industrialization potential that can be used for portable diagnostics. PMID:26556354

  10. Electrical microfluidic pressure gauge for elastomer microelectromechanical systems

    PubMed Central

    Kartalov, Emil P.; Maltezos, George; Anderson, W. French; Taylor, Clive R.; Scherer, Axel

    2009-01-01

    We report on an electrical microfluidic pressure gauge. A polydimethylsiloxane microvalve closes at a characteristic applied pressure determined by the material’s properties and the valve’s dimensions. Hence, when the same pressure is applied to all valves of a heterogeneous valve array, some valves close while others remain open. The state of the array is combined with knowledge of the respective characteristic closing pressures of the individual valves to yield an estimate of the applied pressure. The state of each valve is obtained by electrical measurements, since the electrical resistance of the respective underlying fluid-filled channel increases by at least two orders of magnitude as the valve closes and its insulating elastomer material interrupts the electrical circuit. The overall system functions as a pressure gauge with electrical readout. This device would be a critical component in active pressure-regulation loops in future integrated microfluidic systems. PMID:19587835

  11. Suspended microfluidics

    PubMed Central

    Casavant, Benjamin P.; Berthier, Erwin; Theberge, Ashleigh B.; Berthier, Jean; Montanez-Sauri, Sara I.; Bischel, Lauren L.; Brakke, Kenneth; Hedman, Curtis J.; Bushman, Wade; Keller, Nancy P.; Beebe, David J.

    2013-01-01

    Although the field of microfluidics has made significant progress in bringing new tools to address biological questions, the accessibility and adoption of microfluidics within the life sciences are still limited. Open microfluidic systems have the potential to lower the barriers to adoption, but the absence of robust design rules has hindered their use. Here, we present an open microfluidic platform, suspended microfluidics, that uses surface tension to fill and maintain a fluid in microscale structures devoid of a ceiling and floor. We developed a simple and ubiquitous model predicting fluid flow in suspended microfluidic systems and show that it encompasses many known capillary phenomena. Suspended microfluidics was used to create arrays of collagen membranes, mico Dots (μDots), in a horizontal plane separating two fluidic chambers, demonstrating a transwell platform able to discern collective or individual cellular invasion. Further, we demonstrated that μDots can also be used as a simple multiplexed 3D cellular growth platform. Using the μDot array, we probed the combined effects of soluble factors and matrix components, finding that laminin mitigates the growth suppression properties of the matrix metalloproteinase inhibitor GM6001. Based on the same fluidic principles, we created a suspended microfluidic metabolite extraction platform using a multilayer biphasic system that leverages the accessibility of open microchannels to retrieve steroids and other metabolites readily from cell culture. Suspended microfluidics brings the high degree of fluidic control and unique functionality of closed microfluidics into the highly accessible and robust platform of open microfluidics. PMID:23729815

  12. Microfluidic cell culture chip with multiplexed medium delivery and efficient cell/scaffold loading mechanisms for high-throughput perfusion 3-dimensional cell culture-based assays.

    PubMed

    Huang, Song-Bin; Wu, Min-Hsien; Wang, Shih-Siou; Lee, Gwo-Bin

    2011-06-01

    This study reports a microfluidic cell culture chip consisting of 48 microbioreactors for high-throughput perfusion 3-dimensional (3-D) cell culture-based assays. Its advantages include the capability for multiplexed and backflow-free medium delivery, and both efficient and high-throughput micro-scale, 3-D cell culture construct loading. In this work, the microfluidic cell culture chip is fabricated using two major processes, specifically, a computer-numerical-controlled (CNC) mold machining process and a polydimethylsiloxane (PDMS) replication process. The chip is composed of micropumps, microbioreactors, connecting microchannels and a cell/agarose scaffold loading mechanism. The performance of the new pneumatic micropumps and the cell/agarose scaffold loading mechanism has been experimentally evaluated. The experimental results show that this proposed multiplexed medium-pumping design is able to provide a uniform pumping rate ranging from 1.5 to 298.3 μl hr(-1) without any fluid backflow and the resultant medium contamination. In addition, the simple cell/agarose loading method has been proven to be able to load the 3-D cell culture construct uniformly and efficiently in all 48 microbioreactors investigated. Furthermore, a micro-scale, perfusion, 3-D cell culture-based assay has been successfully demonstrated using this proposed cell culture chip. The experimental results are also compared to a similar evaluation using a conventional static 3-D cell culture with a larger scale culture. It is concluded that the choice of a cell culture format can influence assay results. As a whole, because of the inherent advantages of a miniaturized perfusion 3-D cell culture assay, the cell culture chip not only can provide a stable, well-defined and more biologically-meaningful culture environment, but it also features a low consumption of research resources. Moreover, due to the integrated medium pumping mechanism and the simple cell/agarose loading method, this chip is

  13. Silicon photonic sensors incorporated in a digital microfluidic system.

    PubMed

    Lerma Arce, Cristina; Witters, Daan; Puers, Robert; Lammertyn, Jeroen; Bienstman, Peter

    2012-12-01

    Label-free biosensing with silicon nanophotonic microring resonator sensors has proven to be an excellent sensing technique for achieving high-throughput and high sensitivity, comparing favorably with other labeled and label-free sensing techniques. However, as in any biosensing platform, silicon nanophotonic microring resonator sensors require a fluidic component which allows the continuous delivery of the sample to the sensor surface. This component is typically based on microchannels in polydimethylsiloxane or other materials, which add cost and complexity to the system. The use of microdroplets in a digital microfluidic system, instead of continuous flows, is one of the recent trends in the field, where microliter- to picoliter-sized droplets are generated, transported, mixed, and split, thereby creating miniaturized reaction chambers which can be controlled individually in time and space. This avoids cross talk between samples or reagents and allows fluid plugs to be manipulated on reconfigurable paths, which cannot be achieved using the more established and more complex technology of microfluidic channels where droplets are controlled in series. It has great potential for high-throughput liquid handling, while avoiding on-chip cross-contamination. We present the integration of two miniaturized technologies: label-free silicon nanophotonic microring resonator sensors and digital microfluidics, providing an alternative to the typical microfluidic system based on microchannels. The performance of this combined system is demonstrated by performing proof-of-principle measurements of glucose, sodium chloride, and ethanol concentrations. These results show that multiplexed real-time detection and analysis, great flexibility, and portability make the combination of these technologies an ideal platform for easy and fast use in any laboratory. PMID:22926129

  14. Microfluidic systems with embedded materials and structures and method thereof

    DOEpatents

    Morse, Jeffrey D.; Rose, Klint A; Maghribi, Mariam; Benett, William; Krulevitch, Peter; Hamilton, Julie; Graff, Robert T.; Jankowski, Alan

    2007-03-06

    Described herein is a process for fabricating microfluidic systems with embedded components in which micron-scale features are molded into the polymeric material polydimethylsiloxane (PDMS). Micromachining is used to create a mold master and the liquid precursors for PDMS are poured over the mold and allowed to cure. The PDMS is then removed form the mold and bonded to another material such as PDMS, glass, or silicon after a simple surface preparation step to form sealed microchannels.

  15. Microfluidics co-culture systems for studying tooth innervation

    PubMed Central

    Pagella, Pierfrancesco; Neto, Estrela; Jiménez-Rojo, Lucia; Lamghari, Meriem; Mitsiadis, Thimios A.

    2014-01-01

    Innervation plays a key role in the development and homeostasis of organs and tissues of the orofacial complex. Among these structures, teeth are peculiar organs as they are not innervated until later stages of development. Furthermore, the implication of neurons in tooth initiation, morphogenesis and differentiation is still controversial. Co-cultures constitute a valuable method to investigate and manipulate the interactions of nerve fibers with their target organs in a controlled and isolated environment. Conventional co-cultures between neurons and their target tissues have already been performed, but these cultures do not offer optimal conditions that are closely mimicking the in vivo situation. Indeed, specific cell populations require different culture media in order to preserve their physiological properties. In this study we evaluate the usefulness of a microfluidics system for co-culturing mouse trigeminal ganglia and developing teeth. This device allows the application of specific media for the appropriate development of both neuronal and dental tissues. The results show that mouse trigeminal ganglia and teeth survive for long culture periods in this microfluidics system, and that teeth maintain the attractive or repulsive effect on trigeminal neurites that has been observed in vivo. Neurites are repealed when co-cultured with embryonic tooth germs, while postnatal teeth exert an attractive effect to trigeminal ganglia-derived neurons. In conclusion, microfluidics system devices provide a valuable tool for studying the behavior of neurons during the development of orofacial tissues and organs, faithfully imitating the in vivo situation. PMID:25202282

  16. Multiplex microfluidic system integrating sequential operations of microalgal lipid production.

    PubMed

    Kwak, Ho Seok; Kim, Jaoon Young Hwan; Na, Sang Cheol; Jeon, Noo Li; Sim, Sang Jun

    2016-02-21

    The unit cost for the production of algal biofuel needs to be reduced in order to be a substitute for fossil fuel. To achieve this goal, the development of a novel system is needed for a rapid screening of numerous microalgal species to isolate superior strains with the highest lipid productivity. Here, we developed a PDMS-based multiplex microfluidic system with eight chambers and micropillar arrays to expedite multiple steps for lipid sample preparation from different microalgal strains. We could rapidly and efficiently perform sequential operations from cell culture to lipid extraction of eight different microalgal strains simultaneously on a single device without harvesting and purification steps, which are labor- and energy-intensive, by the simple injection of medium and solvent into the central inlet due to the integrated micropillar arrays connecting the chambers and central inlet. The lipid extraction efficiency using this system was comparable (94.5-102.6%) to the conventional Bligh-Dyer method. We investigated the cell growth and lipid productivity of different strains using the microfluidic device. We observed that each strain has a different lipid accumulation pattern according to stress conditions. These results demonstrate that our multiplex microfluidic approach can provide an efficient analytical tool for the rapid analysis of strain performances (e.g. cell growth and lipid productivities) and the determination of the optimal lipid induction condition for each strain. PMID:26783562

  17. Evaluation of peristaltic micromixers for highly integrated microfluidic systems

    NASA Astrophysics Data System (ADS)

    Kim, Duckjong; Rho, Hoon Suk; Jambovane, Sachin; Shin, Soojeong; Hong, Jong Wook

    2016-03-01

    Microfluidic devices based on the multilayer soft lithography allow accurate manipulation of liquids, handling reagents at the sub-nanoliter level, and performing multiple reactions in parallel processors by adapting micromixers. Here, we have experimentally evaluated and compared several designs of micromixers and operating conditions to find design guidelines for the micromixers. We tested circular, triangular, and rectangular mixing loops and measured mixing performance according to the position and the width of the valves that drive nanoliters of fluids in the micrometer scale mixing loop. We found that the rectangular mixer is best for the applications of highly integrated microfluidic platforms in terms of the mixing performance and the space utilization. This study provides an improved understanding of the flow behaviors inside micromixers and design guidelines for micromixers that are critical to build higher order fluidic systems for the complicated parallel bio/chemical processes on a chip.

  18. Microfluidic hubs, systems, and methods for interface fluidic modules

    DOEpatents

    Bartsch, Michael S; Claudnic, Mark R; Kim, Hanyoup; Patel, Kamlesh D; Renzi, Ronald F; Van De Vreugde, James L

    2015-01-27

    Embodiments of microfluidic hubs and systems are described that may be used to connect fluidic modules. A space between surfaces may be set by fixtures described herein. In some examples a fixture may set substrate-to-substrate spacing based on a distance between registration surfaces on which the respective substrates rest. Fluidic interfaces are described, including examples where fluid conduits (e.g. capillaries) extend into the fixture to the space between surfaces. Droplets of fluid may be introduced to and/or removed from microfluidic hubs described herein, and fluid actuators may be used to move droplets within the space between surfaces. Continuous flow modules may be integrated with the hubs in some examples.

  19. Evaluation of peristaltic micromixers for highly integrated microfluidic systems.

    PubMed

    Kim, Duckjong; Rho, Hoon Suk; Jambovane, Sachin; Shin, Soojeong; Hong, Jong Wook

    2016-03-01

    Microfluidic devices based on the multilayer soft lithography allow accurate manipulation of liquids, handling reagents at the sub-nanoliter level, and performing multiple reactions in parallel processors by adapting micromixers. Here, we have experimentally evaluated and compared several designs of micromixers and operating conditions to find design guidelines for the micromixers. We tested circular, triangular, and rectangular mixing loops and measured mixing performance according to the position and the width of the valves that drive nanoliters of fluids in the micrometer scale mixing loop. We found that the rectangular mixer is best for the applications of highly integrated microfluidic platforms in terms of the mixing performance and the space utilization. This study provides an improved understanding of the flow behaviors inside micromixers and design guidelines for micromixers that are critical to build higher order fluidic systems for the complicated parallel bio/chemical processes on a chip. PMID:27036809

  20. Flow control for capillary-pumped microfluidic systems

    NASA Astrophysics Data System (ADS)

    Vestad, T.; Marr, D. W. M.; Oakey, J.

    2004-11-01

    Advantages of performing analytical and diagnostic tasks in microfluidic-based systems include small sample volume requirements, rapid transport times and the promise of compact, portable instrumentation. The application of such systems in home and point-of-care situations has been limited, however, because these devices typically require significant associated hardware to initiate and control fluid flow. Capillary-based pumping can address many of these deficiencies by taking advantage of surface tension to pull fluid through devices. The development of practical instrumentation however will rely upon the development of precision control schemes to complement capillary pumping. Here, we introduce a straightforward, robust approach that allows for reconfigurable fluid guidance through otherwise fixed capillary networks. This technique is based on the opening and closing of microfluidic channels cast in a flexible elastomer via automated or even manual mechanical actuation. This straightforward approach can completely and precisely control flows such as samples of complex fluids, including whole blood, at very high resolutions according to real-time user feedback. These results demonstrate the suitability of this technique for portable, microfluidic instruments in laboratory, field or clinical diagnostic applications.

  1. Label-free cell separation and sorting in microfluidic systems

    PubMed Central

    Gossett, Daniel R.; Weaver, Westbrook M.; Mach, Albert J.; Hur, Soojung Claire; Tse, Henry Tat Kwong; Lee, Wonhee; Amini, Hamed

    2010-01-01

    Cell separation and sorting are essential steps in cell biology research and in many diagnostic and therapeutic methods. Recently, there has been interest in methods which avoid the use of biochemical labels; numerous intrinsic biomarkers have been explored to identify cells including size, electrical polarizability, and hydrodynamic properties. This review highlights microfluidic techniques used for label-free discrimination and fractionation of cell populations. Microfluidic systems have been adopted to precisely handle single cells and interface with other tools for biochemical analysis. We analyzed many of these techniques, detailing their mode of separation, while concentrating on recent developments and evaluating their prospects for application. Furthermore, this was done from a perspective where inertial effects are considered important and general performance metrics were proposed which would ease comparison of reported technologies. Lastly, we assess the current state of these technologies and suggest directions which may make them more accessible. Figure A wide range of microfluidic technologies have been developed to separate and sort cells by taking advantage of differences in their intrinsic biophysical properties PMID:20419490

  2. DIELECTROPHORESIS-BASED MICROFLUIDIC SEPARATION AND DETECTION SYSTEMS

    PubMed Central

    Yang, Jun; Vykoukal, Jody; Noshari, Jamileh; Becker, Frederick; Gascoyne, Peter; Krulevitch, Peter; Fuller, Chris; Ackler, Harold; Hamilton, Julie; Boser, Bernhard; Eldredge, Adam; Hitchens, Duncan; Andrews, Craig

    2009-01-01

    Diagnosis and treatment of human diseases frequently requires isolation and detection of certain cell types from a complex mixture. Compared with traditional separation and detection techniques, microfluidic approaches promise to yield easy-to-use diagnostic instruments tolerant of a wide range of operating environments and capable of accomplishing automated analyses. These approaches will enable diagnostic advances to be disseminated from sophisticated clinical laboratories to the point-of-care. Applications will include the separation and differential analysis of blood cell subpopulations for host-based detection of blood cell changes caused by disease, infection, or exposure to toxins, and the separation and analysis of surface-sensitized, custom dielectric beads for chemical, biological, and biomolecular targets. Here we report a new particle separation and analysis microsystem that uses dielectrophoretic field-flow fractionation (DEP-FFF). The system consists of a microfluidic chip with integrated sample injector, a DEP-FFF separator, and an AC impedance sensor. We show the design of a miniaturized impedance sensor integrated circuit (IC) with improved sensitivity, a new packaging approach for micro-flumes that features a slide-together compression package and novel microfluidic interconnects, and the design, control, integration and packaging of a fieldable prototype. Illustrative applications will be shown, including the separation of different sized beads and different cell types, blood cell differential analysis, and impedance sensing results for beads, spores and cells. PMID:22025905

  3. Rapid laser prototyping of valves for microfluidic autonomous systems

    NASA Astrophysics Data System (ADS)

    Mohammed, M. I.; Abraham, E.; Y Desmulliez, M. P.

    2013-03-01

    Capillary forces in microfluidics provide a simple yet elegant means to direct liquids through flow channel networks. The ability to manipulate the flow in a truly automated manner has proven more problematic. The majority of valves require some form of flow control devices, which are manually, mechanically or electrically driven. Most demonstrated capillary systems have been manufactured by photolithography, which, despite its high precision and repeatability, can be labour intensive, requires a clean room environment and the use of fixed photomasks, limiting thereby the agility of the manufacturing process to readily examine alternative designs. In this paper, we describe a robust and rapid CO2 laser manufacturing process and demonstrate a range of capillary-driven microfluidic valve structures embedded within a microfluidic network. The manufacturing process described allows for advanced control and manipulation of fluids such that flow can be halted, triggered and delayed based on simple geometrical alterations to a given microchannel. The rapid prototyping methodology has been employed with PMMA substrates and a complete device has been created, ready for use, within 2-3 h. We believe that this agile manufacturing process can be applied to produce a range of complex autonomous fluidic platforms and allows subsequent designs to be rapidly explored.

  4. Development of a fast thermal response microfluidic system using liquid metal

    NASA Astrophysics Data System (ADS)

    Gao, Meng; Gui, Lin

    2016-07-01

    Room temperature liquid metal gallium alloy has been widely used in many micro-electromechanical systems applications, such as on-chip electrical microheaters, micro temperature sensors, micro pumps and so on. Injecting liquid metal into microchannels can provide a simple, rapid, low-cost but efficient way to integrate these elements in microfluidic chips with high accuracy. The liquid metal-filled microstructures can be designed in any shape and easily integrated into microfluidic chips. In this paper, an on-chip liquid metal-based thermal microfluidic system is proposed for quick temperature control at the microscale. The micro system utilizes just one microfluidic chip as a basic working platform, which has liquid metal-based on-chip heaters, temperature sensors and electroosmotic flow pumps. Under the comprehensive control of these elements, the micro system can quickly change the temperature of a target fluid in the microfluidic chip. These liquid metal-based on-chip elements are very helpful for the fabrication and miniaturization of the microfluidic chip. In this paper, deionized water is used to test the temperature control performance of the thermal microfluidic system. According to the experimental results, the micro system can efficiently control the temperature of water ranging from 28 °C to 90 °C. The thermal microfluidic system has great potential for use in many microfluidic applications, such as on-chip polymerase chain reaction, temperature gradient focusing, protein crystallization and chemical synthesis.

  5. SmartBuild-a truly plug-n-play modular microfluidic system.

    PubMed

    Yuen, Po Ki

    2008-08-01

    In this Technical Note, for the first time, a truly "plug-n-play" modular microfluidic system (SmartBuild Plug-n-Play Modular Microfluidic System) is presented for designing and building integrated modular microfluidic systems for biological and chemical applications. The modular microfluidic system can be built by connecting multiple microfluidic components together to form a larger integrated system. The SmartBuild System comprises of a motherboard with interconnect channels/grooves, fitting components, microchannel inserts with different configurations and microchips/modules with different functionalities. Also, heaters, micropumps and valving systems can be designed and used in the system. Examples of an integrated mixing system and reaction systems are presented here to demonstrate the versatility of the SmartBuild System. PMID:18651081

  6. Temperature control system for water-perfused suits

    NASA Technical Reports Server (NTRS)

    Brengelmann, G. L.; Mckeag, M.; Rowell, L. B.

    1977-01-01

    A system used to control skin temperature in human subjects wearing water-perfused garments is described. It supplies 8 l/min at 10 psi with water temperature controlled within plus or minus 0.1 C. Temperature control is facilitated by a low circulating thermal mass and a fast responding heater based on a commercially available quartz heat lamp. The system is open so that hot or cold water can be added from the building mains to produce rates of change of water temperature exceeding 5 C/min. These capabilities allow semiautomatic control of skin temperature within plus or minus 0.1 C of desired wave forms.

  7. Microfluidic electronics.

    PubMed

    Cheng, Shi; Wu, Zhigang

    2012-08-21

    Microfluidics, a field that has been well-established for several decades, has seen extensive applications in the areas of biology, chemistry, and medicine. However, it might be very hard to imagine how such soft microfluidic devices would be used in other areas, such as electronics, in which stiff, solid metals, insulators, and semiconductors have previously dominated. Very recently, things have radically changed. Taking advantage of native properties of microfluidics, advances in microfluidics-based electronics have shown great potential in numerous new appealing applications, e.g. bio-inspired devices, body-worn healthcare and medical sensing systems, and ergonomic units, in which conventional rigid, bulky electronics are facing insurmountable obstacles to fulfil the demand on comfortable user experience. Not only would the birth of microfluidic electronics contribute to both the microfluidics and electronics fields, but it may also shape the future of our daily life. Nevertheless, microfluidic electronics are still at a very early stage, and significant efforts in research and development are needed to advance this emerging field. The intention of this article is to review recent research outcomes in the field of microfluidic electronics, and address current technical challenges and issues. The outlook of future development in microfluidic electronic devices and systems, as well as new fabrication techniques, is also discussed. Moreover, the authors would like to inspire both the microfluidics and electronics communities to further exploit this newly-established field. PMID:22711057

  8. Microfluidic System Based High Throughput Drug Screening System for Curcumin/TRAIL Combinational Chemotherapy in Human Prostate Cancer PC3 Cells

    PubMed Central

    An, Dami; Kim, Kwangmi; Kim, Jeongyun

    2014-01-01

    We have developed a fully automated high throughput drug screening (HTDS) system based on the microfluidic cell culture array to perform combinational chemotherapy. This system has 64 individually addressable cell culture chambers where the sequential combinatorial concentrations of two different drugs can be generated by two microfluidic diffusive mixers. Each diffusive mixer has two integrated micropumps connected to the media and the drug reservoirs respectively for generating the desired combination without the need for any extra equipment to perfuse the solution such as syringe pumps. The cell array is periodically exposed to the drug combination with the programmed LabVIEW system during a couple of days without extra handling after seeding the cells into the microfluidic device and also, this device does not require the continuous generation of solutions compared to the previous systems. Therefore, the total amount of drug being consumed per experiment is less than a few hundred micro liters in each reservoir. The utility of this system is demonstrated through investigating the viability of the prostate cancer PC3 cell line with the combinational treatments of curcumin and tumor necrosis factor-alpha related apoptosis inducing ligand (TRAIL). Our results suggest that the system can be used for screening and optimizing drug combination with a small amount of reagent for combinatorial chemotherapy against cancer cells. PMID:25143816

  9. Microfluidic system for single cell sorting with optical tweezers

    NASA Astrophysics Data System (ADS)

    Bruns, Thomas; Becsi, Laszlo; Talkenberg, Marc; Wagner, Michael; Weber, Petra; Mescheder, Ulrich; Schneckenburger, Herbert

    2010-11-01

    A microfluidic system was developed and combined with optical tweezers for single cell sorting. This system consists of a glass chip of 300 μm thickness with an etched crosswise channel structure, a silicon layer for sealing and a PMMA substrate for tubular coupling. Selected cells are trapped and moved in perpendicular direction to the main flow for recovery in special reservoirs and further evaluation (e.g. by polymerase chain reaction, PCR). In addition, maximum light doses and exposure times for maintaining cell viability were determined.

  10. Digital Microfluidic Logic Gates

    NASA Astrophysics Data System (ADS)

    Zhao, Yang; Xu, Tao; Chakrabarty, Krishnendu

    Microfluidic computing is an emerging application for microfluidics technology. We propose microfluidic logic gates based on digital microfluidics. Using the principle of electrowetting-on-dielectric, AND, OR, NOT and XOR gates are implemented through basic droplet-handling operations such as transporting, merging and splitting. The same input-output interpretation enables the cascading of gates to create nontrivial computing systems. We present a potential application for microfluidic logic gates by implementing microfluidic logic operations for on-chip HIV test.

  11. An easy-to-use microfluidic interconnection system to create quick and reversibly interfaced simple microfluidic devices

    NASA Astrophysics Data System (ADS)

    Pfreundt, Andrea; Brandt Andersen, Karsten; Dimaki, Maria; Svendsen, Winnie E.

    2015-11-01

    The presented microfluidic interconnection system provides an alternative for the individual interfacing of simple microfluidic devices fabricated in polymers such as polymethylmethacrylate, polycarbonate and cyclic olefin polymer. A modification of the device inlet enables the direct attachment of tubing (such as polytetrafluoroethylene tubing) secured and sealed by using a small plug, without the need for additional assembly, glue or o-rings. This provides a very clean connection that does not require additional, potentially incompatible, materials. The tightly sealed connection can withstand pressures above 250 psi and therefore supports applications with high flow rates or highly viscous fluids. The ease of incorporation, configuration, fabrication and use make this interconnection system ideal for the rapid prototyping of simple microfluidic devices or other integrated systems that require microfluidic interfaces. It provides a valuable addition to the toolbox of individual and small arrays of connectors suitable for micromachined or template-based injection molded devices since it does not require protruding, threaded or glued modifications on the inlet and avoids bulky and expensive fittings.

  12. A microfluidic mixing system for single-molecule measurements.

    PubMed

    Pfeil, Shawn H; Wickersham, Charles E; Hoffmann, Armin; Lipman, Everett A

    2009-05-01

    This article describes the design and fabrication of a microfluidic mixing system optimized for ultrasensitive optical measurements. Channels are replica-molded in polydimethylsiloxane elastomer and sealed with fused-silica coverglass. The resulting devices have broad chemical compatibility and extremely low fluorescence background, enabling measurements of individual molecules under well-characterized nonequilibrium conditions. Fluid delivery and pressure connections are made using an interface that allows for rapid assembly, rapid sample exchange, and modular device replacement while providing access for high numerical aperture optics. PMID:19485532

  13. Silicon micromachined pumps employing piezoelectric membrane actuation for microfluidic systems

    NASA Astrophysics Data System (ADS)

    Koch, Michael

    Microsystems technology is a rapidly expanding area that comprises electronics, mechanics and optics. In this field, physical/chemical sensing, fluid handling and optical communication are emerging as potential markets. Microfluidic systems like an implantable insulin pump, a drug delivery system and a total chemical analysis system are currently being developed by academia and industry around the world. This project contributes to the area of microfluidics in that a novel thick-film-on-silicon membrane actuator has been developed to allow inexpensive mass production of micropumps. To date piezoelectric plates have been surface mounted onto a silicon membrane. This single chip fabrication method can now be replaced by screen printing thick piezoelectric layers onto 4 inch silicon substrates. Two different pump types have been developed. These are membrane pumps with either cantilever valves or diffuser/nozzle valves. Pump rates between 100 and 200 μl min-1 and backpressures up to 4 kPa have been achieved with these pumps. Along with the technology of micropumps, simulators have been developed. A novel coupled FEM-CFD solver was realised by a computer controlled coupling of two commercially available packages (ANSYS and CFX-Flow3D). The results of this simulator were in good agreement with measurements on micromachined cantilever valves. CFX- Flow3D was also used to successfully model the behaviour of the diffuser/nozzle valve. Finally, the pump has been simulated using a continuity equation. A behavioural dynamic extension of the cantilever valve was necessary to achieve better prediction of the pump rates for higher frequencies. As well, a common process has been developed for microfluidic devices like micromixers, particle counters and sorters as well as flow sensors. The micromixer has been tested already and achieves mixing for input pressures between 2 and 7 kPa. This agrees with simulations of the diffusive mixing with CFX-Flow3D. Together with the micropump

  14. Microfluidic systems and methods of transport and lysis of cells and analysis of cell lysate

    DOEpatents

    Culbertson, Christopher T.; Jacobson, Stephen C.; McClain, Maxine A.; Ramsey, J. Michael

    2004-08-31

    Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

  15. Microfluidic systems and methods for transport and lysis of cells and analysis of cell lysate

    DOEpatents

    Culbertson, Christopher T [Oak Ridge, TN; Jacobson, Stephen C [Knoxville, TN; McClain, Maxine A [Knoxville, TN; Ramsey, J Michael [Knoxville, TN

    2008-09-02

    Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

  16. Development of Microfluidic Systems Enabling High-Throughput Single-Cell Protein Characterization.

    PubMed

    Fan, Beiyuan; Li, Xiufeng; Chen, Deyong; Peng, Hongshang; Wang, Junbo; Chen, Jian

    2016-01-01

    This article reviews recent developments in microfluidic systems enabling high-throughput characterization of single-cell proteins. Four key perspectives of microfluidic platforms are included in this review: (1) microfluidic fluorescent flow cytometry; (2) droplet based microfluidic flow cytometry; (3) large-array micro wells (microengraving); and (4) large-array micro chambers (barcode microchips). We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on three key performance parameters (absolute quantification, sensitivity, and throughput). PMID:26891303

  17. Development of Microfluidic Systems Enabling High-Throughput Single-Cell Protein Characterization

    PubMed Central

    Fan, Beiyuan; Li, Xiufeng; Chen, Deyong; Peng, Hongshang; Wang, Junbo; Chen, Jian

    2016-01-01

    This article reviews recent developments in microfluidic systems enabling high-throughput characterization of single-cell proteins. Four key perspectives of microfluidic platforms are included in this review: (1) microfluidic fluorescent flow cytometry; (2) droplet based microfluidic flow cytometry; (3) large-array micro wells (microengraving); and (4) large-array micro chambers (barcode microchips). We examine the advantages and limitations of each technique and discuss future research opportunities by focusing on three key performance parameters (absolute quantification, sensitivity, and throughput). PMID:26891303

  18. Label-free all-electronic biosensing in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Stanton, Michael A.

    Label-free, all-electronic detection techniques offer great promise for advancements in medical and biological analysis. Electrical sensing can be used to measure both interfacial and bulk impedance changes in conducting solutions. Electronic sensors produced using standard microfabrication processes are easily integrated into microfluidic systems. Combined with the sensitivity of radiofrequency electrical measurements, this approach offers significant advantages over competing biological sensing methods. Scalable fabrication methods also provide a means of bypassing the prohibitive costs and infrastructure associated with current technologies. We describe the design, development and use of a radiofrequency reflectometer integrated into a microfluidic system towards the specific detection of biologically relevant materials. We developed a detection protocol based on impedimetric changes caused by the binding of antibody/antigen pairs to the sensing region. Here we report the surface chemistry that forms the necessary capture mechanism. Gold-thiol binding was utilized to create an ordered alkane monolayer on the sensor surface. Exposed functional groups target the N-terminus, affixing a protein to the monolayer. The general applicability of this method lends itself to a wide variety of proteins. To demonstrate specificity, commercially available mouse anti- Streptococcus Pneumoniae monoclonal antibody was used to target the full-length recombinant pneumococcal surface protein A, type 2 strain D39 expressed by Streptococcus Pneumoniae. We demonstrate the RF response of the sensor to both the presence of the surface decoration and bound SPn cells in a 1x phosphate buffered saline solution. The combined microfluidic sensor represents a powerful platform for the analysis and detection of cells and biomolecules.

  19. Microfluidic filtration system to isolate extracellular vesicles from blood.

    PubMed

    Davies, Ryan T; Kim, Junho; Jang, Su Chul; Choi, Eun-Jeong; Gho, Yong Song; Park, Jaesung

    2012-12-21

    Extracellular vesicles are released by various cell types, particularly tumor cells, and may be potential targets for blood-based cancer diagnosis. However, studies performed on blood-borne vesicles to date have been limited by lack of effective, standardized purification strategies. Using in situ prepared nanoporous membranes, we present a simple strategy employing a microfluidic filtration system to isolate vesicles from whole blood samples. This method can be applied to purify nano-sized particles from blood allowing isolation of intact extracellular vesicles, avoiding the need for laborious and potentially damaging centrifugation steps or overly specific antibody-based affinity purification. Porous polymer monoliths were integrated as membranes into poly(methyl methacrylate) microfluidic chips by benchtop UV photopolymerization through a mask, allowing precise positioning of membrane elements while preserving simplicity of device preparation. Pore size could be manipulated by changing the ratio of porogenic solvent to prepolymer solution, and was tuned to a size proper for extraction of vesicles. Using the membrane as a size exclusion filter, we separated vesicles from cells and large debris by injecting whole blood under pressure through the microfluidic device. To enhance isolation purity, DC electrophoresis was employed as an alternative driving force to propel particles across the filter and increase the separation efficiency of vesicles from proteins. From the whole blood of melanoma-grown mice, we isolated extracellular vesicles and performed RT-PCR to verify their contents of RNA. Melan A mRNA derived from melanoma tumor cells were found enriched in filtered samples, confirming the recovery of vesicles via their cargo. This filtration system can be incorporated into other on-chip processes enabling integrated sample preparation for the downstream analysis of blood-based extracellular vesicles. PMID:23111789

  20. Laminated plastic microfluidic components for biological and chemical systems

    SciTech Connect

    Martin, P.M.; Matson, D.W.; Bennett, W.D.; Lin, Y.; Hammerstrom, D.J.

    1999-07-01

    Laminated plastic microfluidic components are being developed for biological testing systems and chemical sensors. Applications include a DNA thermal cycler, DNA analytical systems, electrophoretic flow systems, dialysis systems, and metal sensors for ground water. This article describes fabrication processes developed for these plastic microfluidic components, and the fabrication of a chromium metal sensor and a microdialysis device. Most of the components have a stacked architecture. Using this architecture, the fluid flows, or is pumped through, as many as nine laminated functional levels. Functions include pumping, mixing, reaction, detection, reservoirs, separations, and electronics. Polyimide, poly(methylmethacrylate) (PMMA), and polycarbonate materials with thicknesses between 25 and 125 {mu}m are used to construct the components. This makes the components low cost, inert to many biological fluids and chemicals, and disposable. The components are fabricated by excimer laser micromachining the microchannel patterns and microstructures in the various laminates. In some cases, micropumps are integrated into these components to move the fluids. Vias and interconnects are also cut by the laser and integrated with micropumps. The laminates are sealed and bonded by adhesive and thermal processes and are leak tight. The parts withstand pressures as high as 790 kPa. Typical channel widths are 50 to 100 {mu}m, with aspect ratios near 5. {copyright} {ital 1999 American Vacuum Society.}

  1. Design, modeling and characterization of microfluidic architectures for high flow rate, small footprint microfluidic systems.

    PubMed

    Saias, Laure; Autebert, Julien; Malaquin, Laurent; Viovy, Jean-Louis

    2011-03-01

    We propose a strategy for optimizing distribution of flow in a microfluidic chamber for microreactor, lateral flow assay and immunocapture applications. It is aimed at maximizing flow throughput, while keeping footprint, cell thickness, and shear stress in the distribution channels at a minimum, and offering a uniform flow field along the whole analysis chamber. In order to minimize footprint, the traditional tree-like or "rhombus" design, in which distribution microchannels undergo a series of splittings into two subchannels with equal lengths and widths, was replaced by a design in which subchannel lengths are unequal, and widths are analytically adapted within the Hele-Shaw approximation, in order to keep the flow resistance uniform along all flow paths. The design was validated by hydrodynamic flow simulation using COMSOL finite element software. Simulations show that, if the channel is too narrow, the Hele-Shaw approximation loses accuracy, and the flow velocity in the chamber can fluctuate by up to 20%. We thus used COMSOL simulation to fine-tune the channel parameters, and obtained a fluctuation of flow velocity across the whole chamber below 10%. The design was then implemented into a PDMS device, and flow profiles were measured experimentally using particle tracking. Finally, we show that this system can be applied to cell sorting in self-assembling magnetic arrays, increasing flow throughput by a factor 100 as compared to earlier reported designs. PMID:21240403

  2. Passive injection control for microfluidic systems

    DOEpatents

    Paul, Phillip H.; Arnold, Don W.; Neyer, David W.

    2004-12-21

    Apparatus for eliminating siphoning, "dead" regions, and fluid concentration gradients in microscale analytical devices. In its most basic embodiment, the present invention affords passive injection control for both electric field-driven and pressure-driven systems by providing additional fluid flow channels or auxiliary channels disposed on either side of a sample separation column. The auxiliary channels are sized such that volumetric fluid flow rate through these channels, while sufficient to move the sample away from the sample injection region in a timely fashion, is less than that through the sample separation channel or chromatograph.

  3. CAD system for automatic analysis of CT perfusion maps

    NASA Astrophysics Data System (ADS)

    Hachaj, T.; Ogiela, M. R.

    2011-03-01

    In this article, authors present novel algorithms developed for the computer-assisted diagnosis (CAD) system for analysis of dynamic brain perfusion, computer tomography (CT) maps, cerebral blood flow (CBF), and cerebral blood volume (CBV). Those methods perform both quantitative analysis [detection and measurement and description with brain anatomy atlas (AA) of potential asymmetries/lesions] and qualitative analysis (semantic interpretation of visualized symptoms). The semantic interpretation (decision about type of lesion: ischemic/hemorrhagic, is the brain tissue at risk of infraction or not) of visualized symptoms is done by, so-called, cognitive inference processes allowing for reasoning on character of pathological regions based on specialist image knowledge. The whole system is implemented in.NET platform (C# programming language) and can be used on any standard PC computer with.NET framework installed.

  4. Zebrafish embryo development in a microfluidic flow-through system.

    PubMed

    Wielhouwer, Eric M; Ali, Shaukat; Al-Afandi, Abdulrahman; Blom, Marko T; Riekerink, Marinus B Olde; Poelma, Christian; Westerweel, Jerry; Oonk, Johannes; Vrouwe, Elwin X; Buesink, Wilfred; vanMil, Harald G J; Chicken, Jonathan; van't Oever, Ronny; Richardson, Michael K

    2011-05-21

    The zebrafish embryo is a small, cheap, whole-animal model which may replace rodents in some areas of research. Unfortunately, zebrafish embryos are commonly cultured in microtitre plates using cell-culture protocols with static buffer replacement. Such protocols are highly invasive, consume large quantities of reagents and do not readily permit high-quality imaging. Zebrafish and rodent embryos have previously been cultured in static microfluidic drops, and zebrafish embryos have also been raised in a prototype polydimethylsiloxane setup in a Petri dish. Other than this, no animal embryo has ever been shown to undergo embryonic development in a microfluidic flow-through system. We have developed and prototyped a specialized lab-on-a-chip made from bonded layers of borosilicate glass. We find that zebrafish embryos can develop in the chip for 5 days, with continuous buffer flow at pressures of 0.005-0.04 MPa. Phenotypic effects were seen, but these were scored subjectively as 'minor'. Survival rates of 100% could be reached with buffer flows of 2 µL per well per min. High-quality imaging was possible. An acute ethanol exposure test in the chip replicated the same assay performed in microtitre plates. More than 100 embryos could be cultured in an area, excluding infrastructure, smaller than a credit card. We discuss how biochip technology, coupled with zebrafish larvae, could allow biological research to be conducted in massive, parallel experiments, at high speed and low cost. PMID:21491052

  5. A hybrid microfluidic platform for cell-based assays via diffusive and convective trans-membrane perfusion

    PubMed Central

    Vereshchagina, Elizaveta; Mc Glade, Declan; Glynn, Macdara; Ducrée, Jens

    2013-01-01

    We present a novel 3D hybrid assembly of a polymer microfluidic chip with polycarbonate track-etched membrane (PCTEM) enabling membrane-supported cell culture. Two chip designs have been developed to establish either diffusive or convective reagent delivery using the integrated PCTEM. While it is well suited to a range of cell-based assays, we specifically employ this platform for the screening of a common antitumor chemotoxic agent (mitomycin C – MMC) on the HL60 myeloid leukemia cell line. The toxic activity of MMC is based on the generation of severe DNA damage in the cells. Using either mode of operation, the HL60 cells were cultured on-chip before, during, and after exposure to MMC at concentrations ranging from 0 to 50 μM. Cell viability was analysed off-chip by the trypan blue dye exclusion assay. The results of the on-chip viability assay were found to be consistent with those obtained off-chip and indicated ca. 40% cell survival at MMC concentration of 50 μM. The catalogue of capabilities of the here described cell assay platform comprises of (i) the culturing of cells either under shear-free conditions or under induced through-membrane flows, (ii) the tight time control of the reagent exposure, (iii) the straightforward assembly of devices, (iv) the flexibility on the choice of the membrane, and, prospectively, (v) the amenability for large-scale parallelization. PMID:24404021

  6. Enhanced optical chromatography in a PDMS microfluidic system

    NASA Astrophysics Data System (ADS)

    Terray, A.; Arnold, J.; Hart, S. J.

    2005-12-01

    The purely refractive index driven separation of uniformly sized polystyrene, n = 1.59 and poly(methylmethacrylate), n = 1.49 in an optical chromatography system has been enhanced through the incorporation of a custom poly(dimethysiloxane) (PDMS) microfluidic system. A customized channel geometry was used to create separate regions with different linear flow velocities tailored to the specific application. These separate flow regions were then used to expose the entities in the separation to different linear flow velocities thus enhancing their separation relative to the same separation in a constant velocity flow environment. A microbiological sample containing spores of the biological warfare agent, Bacillus anthracis, and a common environmental interferent, mulberry pollen, was investigated to test the use of tailored velocity regions. These very different samples were analyzed simultaneously only through the use of tailored velocity regions.

  7. Perfusion Electronic Record Documentation Using Epic Systems Software.

    PubMed

    Riley, Jeffrey B; Justison, George A

    2015-12-01

    The authors comment on Steffens and Gunser's article describing the University of Wisconsin adoption of the Epic anesthesia record to include perfusion information from the cardiopulmonary bypass patient experience. We highlight the current-day lessons and the valuable quality and safety principles the Wisconsin-Epic model anesthesia-perfusion record provides. PMID:26834289

  8. Microfluidic System Simulation Including the Electro-Viscous Effect

    NASA Technical Reports Server (NTRS)

    Rojas, Eileen; Chen, C. P.; Majumdar, Alok

    2007-01-01

    This paper describes a practical approach using a general purpose lumped-parameter computer program, GFSSP (Generalized Fluid System Simulation Program) for calculating flow distribution in a network of micro-channels including electro-viscous effects due to the existence of electrical double layer (EDL). In this study, an empirical formulation for calculating an effective viscosity of ionic solutions based on dimensional analysis is described to account for surface charge and bulk fluid conductivity, which give rise to electro-viscous effect in microfluidics network. Two dimensional slit micro flow data was used to determine the model coefficients. Geometry effect is then included through a Poiseuille number correlation in GFSSP. The bi-power model was used to calculate flow distribution of isotropically etched straight channel and T-junction microflows involving ionic solutions. Performance of the proposed model is assessed against experimental test data.

  9. On-demand droplet release for droplet-based microfluidic system.

    PubMed

    Wang, Wei; Yang, Chun; Liu, YingShuai; Li, Chang Ming

    2010-03-01

    On-demand droplet release from microwell was successfully implemented and well combined with droplet trapping/fusion functions to make an ideal and integrated droplet based microfluidic system. PMID:20162230

  10. Modular integration of electronics and microfluidic systems using flexible printed circuit boards.

    PubMed

    Wu, Amy; Wang, Lisen; Jensen, Erik; Mathies, Richard; Boser, Bernhard

    2010-02-21

    Microfluidic systems offer an attractive alternative to conventional wet chemical methods with benefits including reduced sample and reagent volumes, shorter reaction times, high-throughput, automation, and low cost. However, most present microfluidic systems rely on external means to analyze reaction products. This substantially adds to the size, complexity, and cost of the overall system. Electronic detection based on sub-millimetre size integrated circuits (ICs) has been demonstrated for a wide range of targets including nucleic and amino acids, but deployment of this technology to date has been limited due to the lack of a flexible process to integrate these chips within microfluidic devices. This paper presents a modular and inexpensive process to integrate ICs with microfluidic systems based on standard printed circuit board (PCB) technology to assemble the independently designed microfluidic and electronic components. The integrated system can accommodate multiple chips of different sizes bonded to glass or PDMS microfluidic systems. Since IC chips and flex PCB manufacturing and assembly are industry standards with low cost, the integrated system is economical for both laboratory and point-of-care settings. PMID:20126694

  11. Cell Separation by Non-Inertial Force Fields in Microfluidic Systems

    PubMed Central

    Tsutsui, Hideaki; Ho, Chih-Ming

    2009-01-01

    Cell and microparticle separation in microfluidic systems has recently gained significant attention in sample preparations for biological and chemical studies. Microfluidic separation is typically achieved by applying differential forces on the target particles to guide them into different paths. This paper reviews basic concepts and novel designs of such microfluidic separators with emphasis on the use of non-inertial force fields, including dielectrophoretic force, optical gradient force, magnetic force, and acoustic primary radiation force. Comparisons of separation performances with discussions on physiological effects and instrumentation issues toward point-of-care devices are provided as references for choosing appropriate separation methods for various applications. PMID:20046897

  12. An integrated microfluidic biochemical detection system for protein analysis with magnetic bead-based sampling capabilities.

    PubMed

    Choi, Jin-Woo; Oh, Kwang W; Thomas, Jennifer H; Heineman, William R; Halsall, H Brian; Nevin, Joseph H; Helmicki, Arthur J; Henderson, H Thurman; Ahn, Chong H

    2002-02-01

    This paper presents the development and characterization of an integrated microfluidic biochemical detection system for fast and low-volume immunoassays using magnetic beads, which are used as both immobilization surfaces and bio-molecule carriers. Microfluidic components have been developed and integrated to construct a microfluidic biochemical detection system. Magnetic bead-based immunoassay, as a typical example of biochemical detection and analysis, has been successfully performed on the integrated microfluidic biochemical analysis system that includes a surface-mounted biofilter and electrochemical sensor on a glass microfluidic motherboard. Total time required for an immunoassay was less than 20 min including sample incubation time, and sample volume wasted was less than 50 microl during five repeated assays. Fast and low-volume biochemical analysis has been successfully achieved with the developed biofilter and immunosensor, which is integrated to the microfluidic system. Such a magnetic bead-based biochemical detection system, described in this paper, can be applied to protein analysis systems. PMID:15100857

  13. Moving droplets between closed and open microfluidic systems.

    PubMed

    Wang, Weiqiang; Jones, Thomas B

    2015-05-21

    In electric-field-mediated droplet microfluidics, there are two distinct architectures - closed systems using parallel-plate electrodes and open systems using coplanar electrodes fabricated on an open substrate. An architecture combining both closed and open systems on a chip would facilitate many of the chemical and biological processes now envisioned for the laboratory on a chip. To accomplish such an integration requires a means to move droplets back and forth between the two. This paper presents an investigation of the requirements for such manipulation of both water and oil droplets. The required wetting conditions for a droplet to cross the open/closed boundary is revealed by a force balance analysis and predictions of this model are compared to experimental results. Water droplets can be moved between closed and open systems by electrowetting actuation; droplet detachment from the upper plate is facilitated by the use of beveled edge. The force model predicts that driving an oil droplet from a closed to an open structure requires an oleophobic surface. This prediction has been tested and confirmed using <100> silicon wafers made oleophobic by re-entrant microstructures etched into the surface. PMID:25850701

  14. Field-effect flow control in a polydimethylsiloxane-based microfluidic system.

    PubMed

    Buch, J S; Wang, P C; DeVoe, D L; Lee, C S

    2001-10-01

    The application of the field-effect for direct control of electroosmosis in a polydimethylsiloxane (PDMS)-based microfluidic system, constructed on a silicon wafer with a 2.0 microm electrically insulating layer of silicon dioxide, is demonstrated. This microfluidic system consists of a 2.0 cm open microchannel fabricated on a PDMS slab, which can reversibly adhere to the silicon wafer to form a hybrid microfluidic device. Aside from mechanically serving as a robust bottom substrate to seal the channel and support the microfluidic system, the silicon wafer is exploited to achieve field-effect flow control by grounding the semiconductive silicon medium. When an electric field is applied through the channel, a radial electric potential gradient is created across the silicon dioxide layer that allows for direct control of the zeta potential and the resulting electroosmotic flow (EOF). By configuring this microfluidic system with two power supplies at both ends of the microchannel, the applied electric potentials can be varied for manipulating the polarity and the magnitude of the radial electric potential gradient across the silicon dioxide layer. At the same time, the longitudinal potential gradient through the microchannel, which is used to induce EOF, is held constant. The results of EOF control in this hybrid microfluidic system are presented for phosphate buffer at pH 3 and pH 5. It is also demonstrated that EOF control can be performed at higher solution pH of 6 and 7.4 by modifying the silicon wafer surface with cetyltrimethylammonium bromide (CTAB) prior to assembly of the hybrid microfluidic system. Results of EOF control from this study are compared with those reported in the literature involving the use of other microfluidic devices under comparable solution conditions. PMID:11700719

  15. Dynamic cell culture: a microfluidic function generator for live cell microscopy.

    PubMed

    Lee, Philip J; Gaige, Terry A; Hung, Paul J

    2009-01-01

    We present a microfluidic system for time-lapsed, live cell microscopy with the ability to control solution exchange via a dynamic flow controller. The application specific microfluidic plates are designed to maintain adherent and non-adherent cell types for multiple days with continuous medium perfusion. Upstream channels with flow controlled via custom software allow the delivery of unique exposure profiles to the cultured cells, such as square waves, step functions, ramps, etc. PMID:19209350

  16. Microfluidics on compliant substrates: recent developments in foldable and bendable devices and system packaging

    NASA Astrophysics Data System (ADS)

    Gray, Bonnie L.

    2012-04-01

    Microfluidics is revolutionizing laboratory methods and biomedical devices, offering new capabilities and instrumentation in multiple areas such as DNA analysis, proteomics, enzymatic analysis, single cell analysis, immunology, point-of-care medicine, personalized medicine, drug delivery, and environmental toxin and pathogen detection. For many applications (e.g., wearable and implantable health monitors, drug delivery devices, and prosthetics) mechanically flexible polymer devices and systems that can conform to the body offer benefits that cannot be achieved using systems based on conventional rigid substrate materials. However, difficulties in implementing active devices and reliable packaging technologies have limited the success of flexible microfluidics. Employing highly compliant materials such as PDMS that are typically employed for prototyping, we review mechanically flexible polymer microfluidic technologies based on free-standing polymer substrates and novel electronic and microfluidic interconnection schemes. Central to these new technologies are hybrid microfabrication methods employing novel nanocomposite polymer materials and devices. We review microfabrication methods using these materials, along with demonstrations of example devices and packaging schemes that employ them. We review these recent developments and place them in the context of the fields of flexible microfluidics and conformable systems, and discuss cross-over applications to conventional rigid-substrate microfluidics.

  17. A latchable thermally activated phase change actuator for microfluidic systems

    NASA Astrophysics Data System (ADS)

    Richter, Christiane; Sachsenheimer, Kai; Rapp, Bastian E.

    2016-03-01

    Complex microfluidic systems often require a high number of individually controllable active components like valves and pumps. In this paper we present the development and optimization of a latchable thermally controlled phase change actuator which uses a solid/liquid phase transition of a phase change medium and the displacement of the liquid phase change medium to change and stabilize the two states of the actuator. Because the phase change is triggered by heat produced with ohmic resistors the used control signal is an electrical signal. In contrast to pneumatically activated membrane valves this concept allows the individual control of several dozen actuators with only two external pressure lines. Within this paper we show the general working principle of the actuator and demonstrate its general function and the scalability of the concept at an example of four actuators. Additionally we present the complete results of our studies to optimize the response behavior of the actuator - the influence of the heating power as well as the used phase change medium on melting and solidifying times.

  18. Evaluation of nanoencapsulated verteporfin's cytotoxicity using a microfluidic system.

    PubMed

    Tokarska, Katarzyna; Bułka, Magdalena; Bazylińska, Urszula; Jastrzębska, Elżbieta; Chudy, Michał; Dybko, Artur; Wilk, Kazimiera A; Brzózka, Zbigniew

    2016-08-01

    A new-generation of nanoencapsulated photosensitizers could be a good solution to perform effective photodynamic therapy (PDT). In this paper, we present physicochemical characterization and cellular investigation of newly prepared long-sustained release oil-core polyelectrolyte nanocarriers loaded with verteporfin (nano VP) in relation to free VP. For this purpose, a macroscale multiwell plates and multifunctional microfluidic system (for three types of cell cultures: monoculture, coculture and mixed culture) were used. A physical analysis of nano VP showed its high stability, monodispersity with unimodal shape and highly positive charge, what made them good candidates for cancer treatment. Biological properties (cellular internalization and uptake as well as cytotoxicity) of nano and free VP were evaluated using both carcinoma (A549) and normal (MRC-5) human lung cells. It was investigated that verteporfin was accumulated in cancer cells preferentially. Low cytotoxicity of the tested photosensitizer was observed in both macro, and microscale. However, in experiments performed in the microsystem, nano VP allowed the reduction of cytotoxic effect, especially in relation to the normal cells. It could result from the specific environment of cell growth in the microsystem which can quite closely mimic the in vivo conditions. Our results suggest that the presented microsystem could be a very useful microtool for testing of new generation of photosensitizers in various configurations of cell cultures, which are difficult to perform in the macroscale. Moreover, the prepared nano VP could be successfully used for further research i.e. evaluation of PDT procedures. PMID:26997162

  19. Direct observation of dispersion and mixing processes in microfluidic systems.

    PubMed

    Takabayashi, Yoshimasa; Fujino, Tatsuya; Korenaga, Takashi

    2008-01-01

    The diffusion phenomena, dispersion and mixing processes of the sample solute (Basic Blue 3 dye and KMnO4 aqueous solutions) were directly observed in laminar flow in glass microchannels. Quasi steady-state UV-visible absorption spectrometry was carried out using CCD camera images of the colored sample dispersion and mixing processes, and the absorbance change (DeltaAbs) was discussed based on the dimensionless parameter, tau which represents the flow time renormalized to the diffusion coefficient and the channel cross section. It was found that DeltaAbs showed almost the same tau dependence, even though the solutions and the microchannel sizes differed in laminar flow, if the microchannel fabrication method was the same. On the basis of this fundamental result, the total microchannel length required for the reaction of 2,3-diaminonaphthalene (DAN) and NO2- at a flow rate of 2 microL min(-1) was calculated, and the obtained value ( approximately 100 mm) showed very good agreement with our previous microchip research. It was concluded that both results were useful for designing the microchannel width, depth and length to control the chemical reaction time in recent microfluidic systems. PMID:18997379

  20. Magnetic microfluidic system for isolation of single cells

    NASA Astrophysics Data System (ADS)

    Mitterboeck, Richard; Kokkinis, Georgios; Berris, Theocharis; Keplinger, Franz; Giouroudi, Ioanna

    2015-06-01

    This paper presents the design and realization of a compact, portable and cost effective microfluidic system for isolation and detection of rare circulating tumor cells (CTCs) in suspension. The innovative aspect of the proposed isolation method is that it utilizes superparamagnetic particles (SMPs) to label CTCs and then isolate those using microtraps with integrated current carrying microconductors. The magnetically labeled and trapped CTCs can then be detected by integrated magnetic microsensors e.g. giant magnetoresistive (GMR) or giant magnetoimpedance (GMI) sensors. The channel and trap dimensions are optimized to protect the cells from shear stress and achieve high trapping efficiency. These intact single CTCs can then be used for additional analysis, testing and patient specific drug screening. Being able to analyze the CTCs metastasis-driving capabilities on the single cell level is considered of great importance for developing patient specific therapies. Experiments showed that it is possible to capture single labeled cells in multiple microtraps and hold them there without permanent electric current and magnetic field.

  1. Microfluidic system for high throughput characterisation of echogenic particles.

    PubMed

    Rademeyer, Paul; Carugo, Dario; Lee, Jeong Yu; Stride, Eleanor

    2015-01-21

    Echogenic particles, such as microbubbles and volatile liquid micro/nano droplets, have shown considerable potential in a variety of clinical diagnostic and therapeutic applications. The accurate prediction of their response to ultrasound excitation is however extremely challenging, and this has hindered the optimisation of techniques such as quantitative ultrasound imaging and targeted drug delivery. Existing characterisation techniques, such as ultra-high speed microscopy provide important insights, but suffer from a number of limitations; most significantly difficulty in obtaining large data sets suitable for statistical analysis and the need to physically constrain the particles, thereby altering their dynamics. Here a microfluidic system is presented that overcomes these challenges to enable the measurement of single echogenic particle response to ultrasound excitation. A co-axial flow focusing device is used to direct a continuous stream of unconstrained particles through the combined focal region of an ultrasound transducer and a laser. Both the optical and acoustic scatter from individual particles are then simultaneously recorded. Calibration of the device and example results for different types of echogenic particle are presented, demonstrating a high throughput of up to 20 particles per second and the ability to resolve changes in particle radius down to 0.1 μm with an uncertainty of less than 3%. PMID:25367757

  2. Microfluidic organs-on-chips.

    PubMed

    Bhatia, Sangeeta N; Ingber, Donald E

    2014-08-01

    An organ-on-a-chip is a microfluidic cell culture device created with microchip manufacturing methods that contains continuously perfused chambers inhabited by living cells arranged to simulate tissue- and organ-level physiology. By recapitulating the multicellular architectures, tissue-tissue interfaces, physicochemical microenvironments and vascular perfusion of the body, these devices produce levels of tissue and organ functionality not possible with conventional 2D or 3D culture systems. They also enable high-resolution, real-time imaging and in vitro analysis of biochemical, genetic and metabolic activities of living cells in a functional tissue and organ context. This technology has great potential to advance the study of tissue development, organ physiology and disease etiology. In the context of drug discovery and development, it should be especially valuable for the study of molecular mechanisms of action, prioritization of lead candidates, toxicity testing and biomarker identification. PMID:25093883

  3. Utilization of the organ care system as ex-vivo lung perfusion after cold storage transportation.

    PubMed

    Mohite, P N; Maunz, O; Popov, A-F; Zych, B; Patil, N P; Simon, A R

    2015-11-01

    The Organ Care System (OCS) allows perfusion and ventilation of the donor lungs under physiological conditions. Ongoing trials to compare preservation with OCS Lung with standard cold storage do not include donor lungs with suboptimal gas exchange and donor lungs treated with OCS following cold storage transportation. We present a case of a 48-yr-old man who received such lungs after cold storage transportation treated with ex-vivo lung perfusion utilizing OCS. PMID:25662732

  4. Point-of-care, portable microfluidic blood analyzer system

    NASA Astrophysics Data System (ADS)

    Maleki, Teimour; Fricke, Todd; Quesenberry, J. T.; Todd, Paul W.; Leary, James F.

    2012-03-01

    Recent advances in MEMS technology have provided an opportunity to develop microfluidic devices with enormous potential for portable, point-of-care, low-cost medical diagnostic tools. Hand-held flow cytometers will soon be used in disease diagnosis and monitoring. Despite much interest in miniaturizing commercially available cytometers, they remain costly, bulky, and require expert operation. In this article, we report progress on the development of a battery-powered handheld blood analyzer that will quickly and automatically process a drop of whole human blood by real-time, on-chip magnetic separation of white blood cells (WBCs), fluorescence analysis of labeled WBC subsets, and counting a reproducible fraction of the red blood cells (RBCs) by light scattering. The whole blood (WB) analyzer is composed of a micro-mixer, a special branching/separation system, an optical detection system, and electronic readout circuitry. A droplet of un-processed blood is mixed with the reagents, i.e. magnetic beads and fluorescent stain in the micro-mixer. Valve-less sorting is achieved by magnetic deflection of magnetic microparticle-labeled WBC. LED excitation in combination with an avalanche photodiode (APD) detection system is used for counting fluorescent WBC subsets using several colors of immune-Qdots, while counting a reproducible fraction of red blood cells (RBC) is performed using a laser light scatting measurement with a photodiode. Optimized branching/channel width is achieved using Comsol Multi-Physics™ simulation. To accommodate full portability, all required power supplies (40v, +/-10V, and +3V) are provided via step-up voltage converters from one battery. A simple onboard lock-in amplifier is used to increase the sensitivity/resolution of the pulse counting circuitry.

  5. Photo inactivation of virus particles in microfluidic capillary systems.

    PubMed

    Ren, Yudan; Crump, Colin M; Mackley, Malcolm M; Li Puma, Gianluca; Reis, Nuno M

    2016-07-01

    It has long been established that UVC light is a very effective method for inactivating pathogens in a fluid, yet the application of UVC irradiation to modern biotechnological processes is limited by the intrinsic short penetration distance of UVC light in optically dense protein solutions. This experimental and numerical study establishes that irradiating a fluid flowing continuously in a microfluidic capillary system, in which the diameter of the capillary is tuned to the depth of penetration of UVC light, uniquely treats the whole volume of the fluid to UVC light, resulting in fast and effective inactivation of pathogens, with particular focus to virus particles. This was demonstrated by inactivating human herpes simplex virus type-1 (HSV-1, a large enveloped virus) on a dense 10% fetal calf serum solution in a range of fluoropolymer capillary systems, including a 0.75 mm and 1.50 mm internal diameter capillaries and a high-throughput MicroCapillary Film with mean hydraulic diameter of 206 μm. Up to 99.96% of HSV-1 virus particles were effectively inactivated with a mean exposure time of up to 10 s, with undetectable collateral damage to solution proteins. The kinetics of virus inactivation matched well the results from a new mathematical model that considers the parabolic flow profile in the capillaries, and showed the methodology is fully predictable and scalable and avoids both the side effect of UVC light to proteins and the dilution of the fluid in current tubular UVC inactivation systems. This is expected to speed up the industrial adoption of non-invasive UVC virus inactivation in clinical biotechnology and biomanufacturing of therapeutic molecules. Biotechnol. Bioeng. 2016;113: 1481-1492. © 2015 Wiley Periodicals, Inc. PMID:26694540

  6. A compact fluorescence detection system integrated with LED and PIN photodetector for microfluidic application

    NASA Astrophysics Data System (ADS)

    Gu, Wenwen; Wen, Zhiyu; Xu, Yi; Wen, Zhongquan; Zeng, Wei

    2011-08-01

    Laser Induced Fluorescence (LIF) is the most sensitivity and widely used detection method in microfluidic chip. However the tranditional LIF detection has the disadvantage of bulky which limits the promotion and widely use of microfluidic chip. A novel compact fluorescence detection system which can be generally used in glass based micro-fluidic chip is proposed. The high-powered light emitting diode (LED) is used as the exciting light source, and multimode optical fiber is also used as transmission channel to transmit exciting light into the detection window. The filter is a specially designed thin film with the thickness of only 2-3 microns and is integrated on the bottom of the glass based microfluidic chip. PIN silicon based chip is chosen as the photodetector, and is integrated on the microfluidic chip by silicon and glass bonding. The integrated microfluidic chip is packaged in a specially designed cavity to shield external stray light. Experiment results show that the best working current for LED is 200mA; the distance between optical fiber tip and detection spot is 1.5mm; the transmittance of the thin film filter of emission light can reach up to 90% at the wavelength of 520nm; the detection limits for FITC labeled Phenylalanine is 10-7mol/L, and the on-line electrophoresis detection is also achieved.

  7. Analysis system for characterisation of simple, low-cost microfluidic components

    NASA Astrophysics Data System (ADS)

    Smith, Suzanne; Naidoo, Thegaran; Nxumalo, Zandile; Land, Kevin; Davies, Emlyn; Fourie, Louis; Marais, Philip; Roux, Pieter

    2014-06-01

    There is an inherent trade-off between cost and operational integrity of microfluidic components, especially when intended for use in point-of-care devices. We present an analysis system developed to characterise microfluidic components for performing blood cell counting, enabling the balance between function and cost to be established quantitatively. Microfluidic components for sample and reagent introduction, mixing and dispensing of fluids were investigated. A simple inlet port plugging mechanism is used to introduce and dispense a sample of blood, while a reagent is released into the microfluidic system through compression and bursting of a blister pack. Mixing and dispensing of the sample and reagent are facilitated via air actuation. For these microfluidic components to be implemented successfully, a number of aspects need to be characterised for development of an integrated point-of-care device design. The functional components were measured using a microfluidic component analysis system established in-house. Experiments were carried out to determine: 1. the force and speed requirements for sample inlet port plugging and blister pack compression and release using two linear actuators and load cells for plugging the inlet port, compressing the blister pack, and subsequently measuring the resulting forces exerted, 2. the accuracy and repeatability of total volumes of sample and reagent dispensed, and 3. the degree of mixing and dispensing uniformity of the sample and reagent for cell counting analysis. A programmable syringe pump was used for air actuation to facilitate mixing and dispensing of the sample and reagent. Two high speed cameras formed part of the analysis system and allowed for visualisation of the fluidic operations within the microfluidic device. Additional quantitative measures such as microscopy were also used to assess mixing and dilution accuracy, as well as uniformity of fluid dispensing - all of which are important requirements towards the

  8. Continuous synthesis of zinc oxide nanoparticles in a microfluidic system for photovoltaic application

    NASA Astrophysics Data System (ADS)

    Kang, Hyun Wook; Leem, Juyoung; Yoon, Sang Youl; Sung, Hyung Jin

    2014-02-01

    This study describes the synthesis of zinc oxide nanoparticles (ZnO NPs) using a microfluidic system. A continuous and efficient synthetic process was developed based on a microfluidic reactor in which was implemented a time pulsed mixing method that had been optimized using numerical simulations and experimental methods. Numerical simulations revealed that efficient mixing conditions could be obtained over the frequency range 5-15 Hz. This system used ethanol solutions containing 30 mM sodium hydroxide (NaOH) or 10 mM dehydrated zinc acetate (Zn(OAc)2) under 5 Hz pulsed conditions, which provided the optimal mixing performance conditions. The ZnO NPs prepared using the microfluidic synthetic system or batch-processed system were validated by several analytical methods, including transmission electron microscopy (TEM), energy dispersive X-ray spectrometry (EDS), X-ray diffraction (XRD), UV/VIS NIR and zeta (ζ) potential analysis. Bulk-heterojunction organic photovoltaic cells were fabricated with the synthesized ZnO NPs to investigate the practicability and compared with batch-process synthesized ZnO NPs. The results showed that microfluidic synthesized ZnO NPs had good preservability and stability in working solution and the synthetic microfluidic system provided a low-cost, environmentally friendly approach to the continuous production of ZnO NPs.

  9. [Micro-droplet characterization and its application for amino acid detection in droplet microfluidic system].

    PubMed

    Yuan, Huiling; Dong, Libing; Tu, Ran; Du, Wenbin; Ji, Shiru; Wang, Qinhong

    2014-01-01

    Recently, the droplet microfluidic system attracts interests due to its high throughput and low cost to detect and screen. The picoliter micro-droplets from droplet microfluidics are uniform with respect to the size and shape, and could be used as monodispensed micro-reactors for encapsulation and detection of single cell or its metabolites. Therefore, it is indispensable to characterize micro-droplet and its application from droplet microfluidic system. We first constructed the custom-designed droplet microfluidic system for generating micro-droplets, and then used the micro-droplets to encapsulate important amino acids such as glutamic acid, phenylalanine, tryptophan or tyrosine to test the droplets' properties, including the stability, diffusivity and bio-compatibility for investigating its application for amino acid detection and sorting. The custom-designed droplet microfluidic system could generate the uniformed micro-droplets with a controllable size between 20 to 50 microm. The micro-droplets could be stable for more than 20 h without cross-contamination or fusion each other. The throughput of detection and sorting of the system is about 600 micro-droplets per minute. This study provides a high-throughput platform for the analysis and screening of amino acid-producing microorganisms. PMID:24818488

  10. How to fabricate robust microfluidic systems for a dollar

    NASA Astrophysics Data System (ADS)

    Lapierre, Florian; Cameron, Neil R.; Oakeshott, John; Peat, Thomas; Zhu, Yonggang

    2013-12-01

    Since the past decade, the interest towards microfluidic devices has sensibly grown due to the wide variety of multidisciplinary applications. One branch of the microfluidic domain consists in the synthesis of various types of emulsions requested by cosmetic, food and biotechnological industries In particular, monodisperse water-in-oil microemulsion synthetised in microfluidic devices are quickly becoming the new generation of emulsions for precise bead control and high surface area. These microemulsions are generally aqueous bioreactors in the form of droplets from 500 nm to 10 μm in diameter, enclosed in an oil environment. An increasing demand for bigger emulsions has led us to investigate new techniques for fabricating fluidic devices allowing a better control over the final size of the droplets. An easy, cheap, reproducible and fast technology for generating emulsions in the range of 100s μm with high throughout (up to mL/h) is reported. Simply using pipette tips and tubing, an innovative microfluidic device was fabricated, able to synthetise water-in-oil emulsions within the range 50 - 500 _μm and double emulsions. These new emulsions are currently used for the synthesis of highly porous polymers beads from High Internal Phase Emulsion (HIPE). These beads will find high potential in 3D cell culture due to their high porosity (up to 90%) and pore size (from 5 to 30μm).

  11. Amperometric quantification based on serial dilution microfluidic systems.

    PubMed

    Stephan, Khaled; Pittet, Patrick; Sigaud, Monique; Renaud, Louis; Vittori, Olivier; Morin, Pierre; Ouaini, Naim; Ferrigno, Rosaria

    2009-03-01

    This paper describes a microfluidic device fabricated in poly(dimethylsiloxane) that was employed to perform amperometric quantifications using on-chip calibration curves and on-chip standard addition methods. This device integrated a network of Au electrodes within a microfluidic structure designed for automatic preparation of a series of solutions containing an electroactive molecule at a concentration linearly decreasing. This device was first characterized by fluorescence microscopy and then evaluated with a model electroactive molecule such as Fe(CN(6))(4-). Operating a quantification in this microfluidic parallel approach rather than in batch mode allows a reduced analysis time to be achieved. Moreover, the microfluidic approach is compatible with the on-chip calibration of sensors simultaneously to the analysis, therefore preventing problems due to sensor response deviation with time. When using the on-chip calibration and on-chip standard addition method, we reached concentration estimation better than 5%. We also demonstrated that compared to the calibration curve approach, the standard addition mode is less complex to operate. Indeed, in this case, it is not necessary to take into account flow rate discrepancies as in the calibration approach. PMID:19238282

  12. Particle dynamics and particle-cell interaction in microfluidic systems

    NASA Astrophysics Data System (ADS)

    Stamm, Matthew T.

    Particle-laden flow in a microchannel resulting in aggregation of microparticles was investigated to determine the dependence of the cluster growth rate on the following parameters: suspension void fraction, shear strain rate, and channel-height to particle-diameter ratio. The growth rate of an average cluster was found to increase linearly with suspension void fraction, and to obey a power-law relationships with shear strain rate as S 0.9 and channel-height to particle-diameter ratio as (h/d )--3.5. Ceramic liposomal nanoparticles and silica microparticles were functionalized with antibodies that act as targeting ligands. The bio-functionality and physical integrity of the cerasomes were characterized. Surface functionalization allows cerasomes to deliver drugs with selectivity and specificity that is not possible using standard liposomes. The functionalized particle-target cell binding process was characterized using BT-20 breast cancer cells. Two microfluidic systems were used; one with both species in suspension, the other with cells immobilized inside a microchannel and particle suspension as the mobile phase. Effects of incubation time, particle concentration, and shear strain rate on particle-cell binding were investigated. With both species in suspension, the particle-cell binding process was found to be reasonably well-described by a first-order model. Particle desorption and cellular loss of binding affinity in time were found to be negligible; cell-particle-cell interaction was identified as the limiting mechanism in particle-cell binding. Findings suggest that separation of a bound particle from a cell may be detrimental to cellular binding affinity. Cell-particle-cell interactions were prevented by immobilizing cells inside a microchannel. The initial stage of particle-cell binding was investigated and was again found to be reasonably well-described by a first-order model. For both systems, the time constant was found to be inversely proportional to

  13. Functional maintenance of differentiated embryoid bodies in microfluidic systems: a platform for personalized medicine.

    PubMed

    Guven, Sinan; Lindsey, Jennifer S; Poudel, Ishwari; Chinthala, Sireesha; Nickerson, Michael D; Gerami-Naini, Behzad; Gurkan, Umut A; Anchan, Raymond M; Demirci, Utkan

    2015-03-01

    Hormone replacement therapies have become important for treating diseases such as premature ovarian failure or menopausal complications. The clinical use of bioidentical hormones might significantly reduce some of the potential risks reportedly associated with the use of synthetic hormones. In the present study, we demonstrate the utility and advantage of a microfluidic chip culture system to enhance the development of personalized, on-demand, treatment modules using embryoid bodies (EBs). Functional EBs cultured on microfluidic chips represent a platform for personalized, patient-specific treatment cassettes that can be cryopreserved until required for treatment. We assessed the viability, differentiation, and functionality of EBs cultured and cryopreserved in this system. During extended microfluidic culture, estradiol, progesterone, testosterone, and anti-müllerian hormone levels were measured, and the expression of differentiated steroidogenic cells was confirmed by immunocytochemistry assay for the ovarian tissue markers anti-müllerian hormone receptor type II, follicle-stimulating hormone receptor, and inhibin β-A and the estrogen biosynthesis enzyme aromatase. Our studies showed that under microfluidic conditions, differentiated steroidogenic EBs continued to secrete estradiol and progesterone at physiologically relevant concentrations (30-120 pg/ml and 150-450 pg/ml, respectively) for up to 21 days. Collectively, we have demonstrated for the first time the feasibility of using a microfluidic chip system with continuous flow for the differentiation and extended culture of functional steroidogenic stem cell-derived EBs, the differentiation of EBs into cells expressing ovarian antigens in a microfluidic system, and the ability to cryopreserve this system with restoration of growth and functionality on thawing. These results present a platform for the development of a new therapeutic system for personalized medicine. PMID:25666845

  14. Functional Maintenance of Differentiated Embryoid Bodies in Microfluidic Systems: A Platform for Personalized Medicine

    PubMed Central

    Guven, Sinan; Lindsey, Jennifer S.; Poudel, Ishwari; Chinthala, Sireesha; Nickerson, Michael D.; Gerami-Naini, Behzad; Gurkan, Umut A.

    2015-01-01

    Hormone replacement therapies have become important for treating diseases such as premature ovarian failure or menopausal complications. The clinical use of bioidentical hormones might significantly reduce some of the potential risks reportedly associated with the use of synthetic hormones. In the present study, we demonstrate the utility and advantage of a microfluidic chip culture system to enhance the development of personalized, on-demand, treatment modules using embryoid bodies (EBs). Functional EBs cultured on microfluidic chips represent a platform for personalized, patient-specific treatment cassettes that can be cryopreserved until required for treatment. We assessed the viability, differentiation, and functionality of EBs cultured and cryopreserved in this system. During extended microfluidic culture, estradiol, progesterone, testosterone, and anti-müllerian hormone levels were measured, and the expression of differentiated steroidogenic cells was confirmed by immunocytochemistry assay for the ovarian tissue markers anti-müllerian hormone receptor type II, follicle-stimulating hormone receptor, and inhibin β-A and the estrogen biosynthesis enzyme aromatase. Our studies showed that under microfluidic conditions, differentiated steroidogenic EBs continued to secrete estradiol and progesterone at physiologically relevant concentrations (30–120 pg/ml and 150–450 pg/ml, respectively) for up to 21 days. Collectively, we have demonstrated for the first time the feasibility of using a microfluidic chip system with continuous flow for the differentiation and extended culture of functional steroidogenic stem cell-derived EBs, the differentiation of EBs into cells expressing ovarian antigens in a microfluidic system, and the ability to cryopreserve this system with restoration of growth and functionality on thawing. These results present a platform for the development of a new therapeutic system for personalized medicine. PMID:25666845

  15. Piezo-microfluidic transport system for multi-targets biochip detections

    NASA Astrophysics Data System (ADS)

    Li, Chia-Chin; Wang, Pei-Wen; Lee, Chih-Kung

    2016-03-01

    Detecting minute trace of interferon-gamma and various bio-markers by using a single biochip was adopted as a platform to examine the technology advancements presented. As bio-detection faces the restriction that only very small quantity of specimen is available, ways to make the best use of the sample available are a must. Since samples concentration will affect the binding rate of an immunoassay, the testing order will become an influencing factor if multiple biomarkers testing situation are needed by using only a single trace of sample. More specifically, if we test disease A first and then detect disease B using the sample just been measured by testing disease A, we most likely will get different results if we reverse the testing order. With an attempt to examine and maybe resolve the issues mentioned above, a micro-fluid control system was developed. The design requirements not only ask for microfluidic control but also demand the system developed has the potential to be integrated within the biochip once its performance is verified. A piezo-vibrating system that can generate traveling waves for microfluidic control was chosen due to its versatility and large force to volume ratio. A simulation software COMSOL was adopted first to predict the microfluidic behavior of the two-mode excited piezo-microfluidic transport system. Secondly, fluorescent particles was used to analyze the microfluidic behavior of system fabricated based on the simulation. Finally, Electrochemistry Impedance Spectroscopy (EIS) was implemented to verify the performance and extendibility of this newly developed system for multi-target detections.

  16. Rapid prototyping of arrayed microfluidic systems in polystyrene for cell-based assays

    PubMed Central

    Young, Edmond W.K.; Berthier, Erwin; Guckenberger, David J.; Sackmann, Eric; Lamers, Casey; Meyvantsson, Ivar; Huttenlocher, Anna; Beebe, David J.

    2011-01-01

    Microfluidic cell-based systems have enabled the study of cellular phenomena with improved spatiotemporal control of the microenvironment and at increased throughput. While PDMS has emerged as the most popular material in microfluidics research, it has specific limitations that prevent microfluidic platforms from achieving their full potential. We present here a complete process, ranging from mold design to embossing and bonding, that describes the fabrication of polystyrene (PS) microfluidic devices with similar cost and time expenditures as PDMS-based devices. Emphasis was placed on creating methods that can compete with PDMS fabrication methods in terms of robustness, complexity and time requirements. To achieve this goal several improvements were made to remove critical bottlenecks in existing PS embossing methods. First, traditional lithography techniques were adapted to fabricate bulk epoxy molds capable of resisting high temperatures and pressures. Second, a method was developed to emboss through-holes in a PS layer, enabling creation of large arrays of independent microfluidic systems on a single device without need to manually create access ports. Third, thermal bonding of PS layers was optimized in order to achieve quality bonding over large arrays of microsystems. The choice of materials and methods were validated for biological function using two different cell-based applications to demonstrate the versatility of our streamlined fabrication process. PMID:21261280

  17. Microfluidic devices, systems, and methods for quantifying particles using centrifugal force

    SciTech Connect

    Schaff, Ulrich Y.; Sommer, Gregory J.; Singh, Anup K.

    2015-11-17

    Embodiments of the present invention are directed toward microfluidic systems, apparatus, and methods for measuring a quantity of cells in a fluid. Examples include a differential white blood cell measurement using a centrifugal microfluidic system. A method may include introducing a fluid sample containing a quantity of cells into a microfluidic channel defined in part by a substrate. The quantity of cells may be transported toward a detection region defined in part by the substrate, wherein the detection region contains a density media, and wherein the density media has a density lower than a density of the cells and higher than a density of the fluid sample. The substrate may be spun such that at least a portion of the quantity of cells are transported through the density media. Signals may be detected from label moieties affixed to the cells.

  18. Interaction of droplets in recirculation regions within microfluidic systems

    NASA Astrophysics Data System (ADS)

    Ghazi, Nastaran; Hosseini, Ashkan; Shojaei-Zadeh, Shahab

    2012-11-01

    We investigate the interaction of oil droplets in continuous water phase as they travel across the streamlines of a recirculation region using microfluidic devices. Oil droplets are first generated using hydrodynamic focusing and then enter a recirculation region. The droplets then keep recirculating until they are pushed out by the incoming ones. We show that the frequency of droplet generation, viscosity contrast (oil to water), and geometry determine which droplets to stay in the recirculation region and which one to leave. Using flow field simulations, we investigate the migration of droplets and their trajectories based on the geometry of the recirculation region, the bubble size, and fluid properties. Under favorable conditions, when droplets interact within the recirculation region for long enough time, the film thickness that separates the two interfaces reduces and droplets will coalesce. The proposed design thus provides a suitable platform to study droplet coalescence within microfluidic devices.

  19. Mobile Monolith Polymer Elements For Flow Control In Microfluidic Systems

    DOEpatents

    Hasselbrink, Jr., Ernest F.; Rehm, Jason E.; Shepodd, Timothy J.; Kirby, Brian J.

    2006-01-24

    A cast-in-place and lithographically shaped mobile, monolithic polymer element for fluid flow control in microfluidic devices and method of manufacture. Microfluid flow control devices, or microvalves that provide for control of fluid or ionic current flow can be made incorporating a cast-in-place, mobile monolithic polymer element, disposed within a microchannel, and driven by fluid pressure (either liquid or gas) against a retaining or sealing surface. The polymer elements are made by the application of lithographic methods to monomer mixtures formulated in such a way that the polymer will not bond to microchannel walls. The polymer elements can seal against pressures greater than 5000 psi, and have a response time on the order of milliseconds. By the use of energetic radiation it is possible to depolymerize selected regions of the polymer element to form shapes that cannot be produced by conventional lithographic patterning and would be impossible to machine.

  20. Real-time optical pH measurement in a standard microfluidic cell culture system.

    PubMed

    Magnusson, Einar B; Halldorsson, Skarphedinn; Fleming, Ronan M T; Leosson, Kristjan

    2013-01-01

    The rapid growth of microfluidic cell culturing in biological and biomedical research and industry calls for fast, non-invasive and reliable methods of evaluating conditions such as pH inside a microfluidic system. We show that by careful calibration it is possible to measure pH within microfluidic chambers with high accuracy and precision, using a direct single-pass measurement of light absorption in a commercially available phenol-red-containing cell culture medium. The measurement is carried out using a standard laboratory microscope and, contrary to previously reported methods, requires no modification of the microfluidic device design. We demonstrate the validity of this method by measuring absorption of light transmitted through 30-micrometer thick microfluidic chambers, using an inverted microscope fitted with a scientific-grade digital camera and two bandpass filters. In the pH range of 7-8, our measurements have a standard deviation and absolute error below 0.05 for a measurement volume smaller than 4 nL. PMID:24049695

  1. Development of microfluidic system and optical tweezers for electrophysiological investigations of an individual cell

    NASA Astrophysics Data System (ADS)

    Alrifaiy, A.; Bitaraf, N.; Lindahl, O.; Ramser, K.

    2010-08-01

    We present a new approach of combining Lab-on-a-chip technologies with optical manipulation technique for accurate investigations in the field of cell biology. A general concept was to develop and combine different methods to perform advanced electrophysiological investigations of an individual living cell under optimal control of the surrounding environment. The conventional patch clamp technique was customized by modifying the open system with a gas-tight multifunctional microfluidics system and optical trapping technique (optical tweezers). The system offers possibilities to measure the electrical signaling and activity of the neuron under optimum conditions of hypoxia and anoxia while the oxygenation state is controlled optically by means of a spectroscopic technique. A cellbased microfluidics system with an integrated patch clamp pipette was developed successfully. Selectively, an individual neuron is manipulated within the microchannels of the microfluidic system under a sufficient control of the environment. Experiments were performed to manipulate single yeast cell and red blood cell (RBC) optically through the microfluidics system toward an integrated patch clamp pipette. An absorption spectrum of a single RCB was recorded which showed that laser light did not impinge on the spectroscopic spectrum of light. This is promising for further development of a complete lab-on-a-chip system for patch clamp measurements.

  2. Zeta potential determination by streaming current modelization and measurement in electrophoretic microfluidic systems.

    PubMed

    Renaud, Louis; Kleimann, Pascal; Morin, Pierre

    2004-01-01

    Electrophoresis in capillary and microfluidic systems, used in analytical chemistry to separate charged species, are quite sensitive to surface phenomena in terms of separation performances. In order to improve theses performances, new surface functionalization techniques are required. There is a need for methods to provide fast and accurate quantification about surface charges at liquid/solid interfaces. We present a fast, simple, and low-cost technique for the measurement of the zeta-potential, via the modelization and the measurement of streaming currents. Due to the small channel cross section in microfluidic devices, the streaming current modelization is easier than the streaming potential measurement. The modelization combines microfluidic simulations based on the Navier-Stokes equation and charge repartition simulations based on the Poisson-Boltzmann equation. This method has been validated with square and circular cross section shape fused-silica capillaries and can be easily transposed to any lab-on-chip microsystems. PMID:14730576

  3. An integrated microfluidic system for bovine DNA purification and digital PCR detection.

    PubMed

    Tian, Qingchang; Mu, Ying; Xu, Yanan; Song, Qi; Yu, Bingwen; Ma, Congcong; Jin, Wei; Jin, Qinhan

    2015-12-15

    In this paper, we described an integrated modularized microfluidic system that contained two distinct functional modules, one for nucleic acids (NA) extraction and the other for digital PCR (dPCR), allowing for detecting the bovine DNA in ovine tissue. PMID:26364950

  4. Screening of C60 crystallization using a microfluidic system.

    PubMed

    Shinohara, Kyosuke; Fukui, Takeshi; Abe, Hiroaki; Sekimura, Naoto; Okamoto, Koji

    2006-07-18

    We have carried out screening of C60 crystallization using a simple liquid/liquid interfacial precipitation method in a microfluidic device. By controlling the time, temperature, and concentration, various metastable phases of C60 crystals were found, including tubes, spheres, open-ended hollow columns, stars, branches, and trees. The obtained C60 crystal shapes are similar to those of snow crystals. These findings suggest an urgent need to screen C60 crystallization for the development of fullerene C60 drugs. PMID:16830986

  5. Tissue culture on a chip: Developmental biology applications of self-organized capillary networks in microfluidic devices.

    PubMed

    Miura, Takashi; Yokokawa, Ryuji

    2016-08-01

    Organ culture systems are used to elucidate the mechanisms of pattern formation in developmental biology. Various organ culture techniques have been used, but the lack of microcirculation in such cultures impedes the long-term maintenance of larger tissues. Recent advances in microfluidic devices now enable us to utilize self-organized perfusable capillary networks in organ cultures. In this review, we will overview past approaches to organ culture and current technical advances in microfluidic devices, and discuss possible applications of microfluidics towards the study of developmental biology. PMID:27272910

  6. Integration of microfluidics/electrochemical system for trace metal analysis by stripping voltammetry

    NASA Astrophysics Data System (ADS)

    Lin, Yuehe; Zhao, Rui; Thrall, Karla D.; Timchalk, C. A.; Bennett, Wendy D.; Matson, Dean W.

    1999-08-01

    Microanalytical systems based on a microfluidics/electrochemical detection scheme were developed. Individual modules, such as microfabricated piezoelectrically actuated pumps and a microelectrochemical cell were integrated onto portable platforms. This allows rapid change-out and repair of individual components by incorporating `plug and play' concepts now standard in PC's. Two different integration schemes were used for construction of the microanalytical systems based on microfluidics/electrochemical detection. In first scheme, all individual modules were integrated in the surface of the standard microfluidic platform based on a plug-and-play design. Microelectrochemical flow cell which integrated three electrodes based on a wall-jet design was fabricated on polymer substrate. The microelectrochemical flow cell was then plugged directly into the microfluidic platform. Another integration scheme was based on a multilayer lamination method utilizing stacking modules with different functionality to achieve a compact microanalytical device. Application of the microanalytical system for detection of lead in river water and saliva samples using stripping voltammetry is described.

  7. A microfluidic system with integrated molecular imprinting polymer films for surface plasmon resonance detection

    NASA Astrophysics Data System (ADS)

    Huang, Shih-Chiang; Lee, Gwo-Bin; Chien, Fan-Ching; Chen, Shean-Jen; Chen, Wen-Janq; Yang, Ming-Chang

    2006-07-01

    This paper presents a novel microfluidic system with integrated molecular imprinting polymer (MIP) films designed for surface plasmon resonance (SPR) biosensing of multiple nanoscale biomolecules. The innovative microfluidic chip uses pneumatic microvalves and micropumps to transport a precise amount of the biosample through multiple microchannels to sensing regions containing the locally spin-coated MIP films. The signals of SPR biosensing are basically proportional to the number of molecules adsorbed on the MIP films. Hence, a precise control of flow rates inside microchannels is important to determine the adsorption amount of the molecules in the SPR/MIP chips. The integration of micropumps and microvalves can automate the sample introduction process and precisely control the amount of the sample injection to the microfluidic system. The proposed biochip enables the label-free biosensing of biomolecules in an automatic format, and provides a highly sensitive, highly specific and high-throughput detection performance. Three samples, i.e. progesterone, cholesterol and testosterone, are successfully detected using the developed system. The experimental results show that the proposed SPR/MIP microfluidic chip provides a comparable sensitivity to that of large-scale SPR techniques, but with reduced sample consumption and an automatic format. As such, the developed biochip has significant potential for a wide variety of nanoscale biosensing applications. The preliminary results of the current paper were presented at Transducers 2005, Seoul, Korea, 5-9 June 2005.

  8. Entropy-based separation of yeast cells using a microfluidic system of conjoined spheres

    SciTech Connect

    Huang, Kai-Jian; Qin, S.-J. Bai, Zhong-Chen; Zhang, Xin; Mai, John D.

    2013-11-21

    A physical model is derived to create a biological cell separator that is based on controlling the entropy in a microfluidic system having conjoined spherical structures. A one-dimensional simplified model of this three-dimensional problem in terms of the corresponding effects of entropy on the Brownian motion of particles is presented. This dynamic mechanism is based on the Langevin equation from statistical thermodynamics and takes advantage of the characteristics of the Fokker-Planck equation. This mechanism can be applied to manipulate biological particles inside a microfluidic system with identical, conjoined, spherical compartments. This theoretical analysis is verified by performing a rapid and a simple technique for separating yeast cells in these conjoined, spherical microfluidic structures. The experimental results basically match with our theoretical model and we further analyze the parameters which can be used to control this separation mechanism. Both numerical simulations and experimental results show that the motion of the particles depends on the geometrical boundary conditions of the microfluidic system and the initial concentration of the diffusing material. This theoretical model can be implemented in future biophysics devices for the optimized design of passive cell sorters.

  9. Laminated microfluidic system for small sample protein analysis

    PubMed Central

    Saedinia, Sara; Nastiuk, Kent L.; Krolewski, John J.; Li, G. P.; Bachman, Mark

    2014-01-01

    We describe a technology based on lamination that allows for the production of highly integrated 3D devices suitable for performing a wide variety of microfluidic assays. This approach uses a suite of microfluidic coupons (“microfloupons”) that are intended to be stacked as needed to produce an assay of interest. Microfloupons may be manufactured in paper, plastic, gels, or other materials, in advance, by different manufacturers, then assembled by the assay designer as needed. To demonstrate this approach, we designed, assembled, and characterized a microfloupon device that performs sodium-dodecyl-sulfate polyacrylamide gel electrophoresis on a small sample of protein. This device allowed for the manipulation and transport of small amounts of protein sample, tight injection into a thin polyacrylamide gel, electrophoretic separation of the proteins into bands, and subsequent removal of the gel from the device for imaging and further analysis. The microfloupons are rugged enough to handle and can be easily aligned and laminated, allowing for a variety of different assays to be designed and configured by selecting appropriate microfloupons. This approach provides a convenient way to perform assays that have multiple steps, relieving the need to design highly sophisticated devices that incorporate all functions in a single unit, while still achieving the benefits of small sample size, automation, and high speed operation. PMID:24753728

  10. Electrokinetic Properties of Lubricin Antiadhesive Coatings in Microfluidic Systems.

    PubMed

    Greene, George W; Duffy, Emer; Shallan, Aliaa; Wuethrich, Alain; Paull, Brett

    2016-02-23

    Lubricin is a glycoprotein found in articular joints which has long been recognized as being an important biological boundary lubricant molecule and, more recently, an impressive antiadhesive that readily self-assembles into a well ordered, polymer brush layer on virtually any substrate. The lubricin molecule possesses an overabundance of anionic charge, a property that is atypical among antiadhesive molecules, that enables its use as a coating for applications involving electrokinetic processes such as electrophoresis and electroosmosis. Coating the surfaces of silica and polymeric microfluidic devices with self-assembled lubricin coatings affords a unique combination of excellent fouling resistance and high charge density that enables notoriously "sticky" biomolecules such as proteins to be used and controlled electrokinetically in the device without complications arising from nonspecific adsorption. Using capillary electrophoresis, we characterized the stability, uniformity, and electrokinetic properties of lubricin coatings applied to silica and PTFE capillaries over a range of run buffer pHs and when exposed to concentrated solutions of protein. In addition, we demonstrate the effectiveness of lubricin as a coating to minimize nonspecific protein adsorption in an electrokinetically controlled polydimethylsiloxane/silica microfluidic device. PMID:26814794

  11. The effect of captopril on thallium 201 myocardial perfusion in systemic sclerosis

    SciTech Connect

    Kahan, A.; Devaux, J.Y.; Amor, B.; Menkes, C.J.; Weber, S.; Venot, A.; Strauch, G. )

    1990-04-01

    In systemic sclerosis, abnormalities of myocardial perfusion are common and may be caused by a disturbance of the coronary microcirculation. We evaluated the long-term effect of captopril (75 to 150 mg per day) on thallium 201 myocardial perfusion in 12 normotensive patients with systemic sclerosis. Captopril significantly decreased the mean (+/- SD) number of segments with thallium 201 myocardial perfusion defects (6.5 +/- 1.9 at baseline and 4.4 +/- 2.7 after 1 year of treatment with captopril; p less than 0.02) and increased the mean global thallium score (9.6 +/- 1.7 at baseline and 11.4 +/- 2.1 after captopril; p less than 0.05). In a control group of eight normotensive patients with systemic sclerosis who did not receive captopril, no significant modification in thallium results occurred. Side effects with captopril included hypotension (six patients), taste disturbances (one patient), and skin rash (one patient). These side effects subsided when the dosage was reduced. These findings demonstrate that captopril improves thallium 201 myocardial perfusion in patients with systemic sclerosis and may therefore have a beneficial effect on scleroderma myocardial disease.

  12. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems

    NASA Astrophysics Data System (ADS)

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-01

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems.

  13. Three-Dimensionally Printed Microfluidic Cross-flow System for Ultrafiltration/Nanofiltration Membrane Performance Testing.

    PubMed

    Wardrip, Nathaniel C; Arnusch, Christopher J

    2016-01-01

    Minimization and management of membrane fouling is a formidable challenge in diverse industrial processes and other practices that utilize membrane technology. Understanding the fouling process could lead to optimization and higher efficiency of membrane based filtration. Here we show the design and fabrication of an automated three-dimensionally (3-D) printed microfluidic cross-flow filtration system that can test up to 4 membranes in parallel. The microfluidic cells were printed using multi-material photopolymer 3-D printing technology, which used a transparent hard polymer for the microfluidic cell body and incorporated a thin rubber-like polymer layer, which prevents leakages during operation. The performance of ultrafiltration (UF), and nanofiltration (NF) membranes were tested and membrane fouling could be observed with a model foulant bovine serum albumin (BSA). Feed solutions containing BSA showed flux decline of the membrane. This protocol may be extended to measure fouling or biofouling with many other organic, inorganic or microbial containing solutions. The microfluidic design is especially advantageous for testing materials that are costly or only available in small quantities, for example polysaccharides, proteins, or lipids due to the small surface area of the membrane being tested. This modular system may also be easily expanded for high throughput testing of membranes. PMID:26968008

  14. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems

    PubMed Central

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-01

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems. PMID:26823292

  15. Integrated optofluidic-microfluidic twin channels: toward diverse application of lab-on-a-chip systems.

    PubMed

    Lv, Chao; Xia, Hong; Guan, Wei; Sun, Yun-Lu; Tian, Zhen-Nan; Jiang, Tong; Wang, Ying-Shuai; Zhang, Yong-Lai; Chen, Qi-Dai; Ariga, Katsuhiko; Yu, Yu-De; Sun, Hong-Bo

    2016-01-01

    Optofluidics, which integrates microfluidics and micro-optical components, is crucial for optical sensing, fluorescence analysis, and cell detection. However, the realization of an integrated system from optofluidic manipulation and a microfluidic channel is often hampered by the lack of a universal substrate for achieving monolithic integration. In this study, we report on an integrated optofluidic-microfluidic twin channels chip fabricated by one-time exposure photolithography, in which the twin microchannels on both surfaces of the substrate were exactly aligned in the vertical direction. The twin microchannels can be controlled independently, meaning that fluids could flow through both microchannels simultaneously without interfering with each other. As representative examples, a tunable hydrogel microlens was integrated into the optofluidic channel by femtosecond laser direct writing, which responds to the salt solution concentration and could be used to detect the microstructure at different depths. The integration of such optofluidic and microfluidic channels provides an opportunity to apply optofluidic detection practically and may lead to great promise for the integration and miniaturization of Lab-on-a-Chip systems. PMID:26823292

  16. A Serial Sample Loading System: Interfacing Multi-well plates with Microfluidic Devices

    PubMed Central

    Rane, Tushar D.; Zec, Helena; Wang, Jeff Tza-Huei

    2013-01-01

    There is an increasing demand for novel high-throughput screening (HTS) technologies in the pharmaceutical and biotechnological industries. The robotic sample handling techniques currently used in these industries, although fast, are still limited to operating in multi-well plates with the sample volumes per reaction in the microliter regime. Digital microfluidics offers an alternative for reduction in sample volume consumption for HTS but lacks a reliable technique for transporting large number of samples to the microfluidic device. In this report, we develop a technique for serial delivery of sample arrays to a microfluidic device from multi-well plates, through a single sample inlet. Under this approach, a serial array of sample plugs, separated by an immiscible carrier fluid, is loaded into a capillary and delivered to a microfluidic device. Similar approaches have been attempted in the past, however, either with a slower sample loading device like syringe pump or vacuum based sample loading with limited driving pressure. We demonstrated the application of our positive pressure based ‘Serial Sample Loading’ (SSL) system to load a series of sample plugs into a capillary. The adaptability of the SSL system to generate sample plugs with a variety of volumes in a predictable manner was also demonstrated. PMID:22885789

  17. Systemic oxygen delivery by peritoneal perfusion of oxygen microbubbles.

    PubMed

    Feshitan, Jameel A; Legband, Nathan D; Borden, Mark A; Terry, Benjamin S

    2014-03-01

    Severe hypoxemia refractory to pulmonary mechanical ventilation remains life-threatening in critically ill patients. Peritoneal ventilation has long been desired for extrapulmonary oxygenation owing to easy access of the peritoneal cavity for catheterization and the relative safety compared to an extracorporeal circuit. Unfortunately, prior attempts involving direct oxygen ventilation or aqueous perfusates of fluorocarbons or hemoglobin carriers have failed, leading many researchers to abandon the method. We attribute these prior failures to limited mass transfer of oxygen to the peritoneum and have designed an oxygen formulation that overcomes this limitation. Using phospholipid-coated oxygen microbubbles (OMBs), we demonstrate 100% survival for rats experiencing acute lung trauma to at least 2 h. In contrast, all untreated rats and rats treated with peritoneal oxygenated saline died within 30 min. For rats treated with OMBs, hemoglobin saturation and heart rate were at normal levels over the 2-h timeframe. Peritoneal oxygenation with OMBs was therefore shown to be safe and effective, and the method requires less equipment and technical expertise than initiating and maintaining an extracorporeal circuit. Further translation of peritoneal oxygenation with OMBs may provide therapy for acute respiratory distress syndrome arising from trauma, sepsis, pneumonia, aspiration, burns and other pulmonary diseases. PMID:24439406

  18. Hydrodynamic guiding for addressing subsets of immobilized cells and molecules in microfluidic systems

    PubMed Central

    Brevig, Thomas; Krühne, Ulrich; Kahn, Rachel A; Ahl, Thomas; Beyer, Michael; Pedersen, Lars H

    2003-01-01

    Background The interest in microfluidics and surface patterning is increasing as the use of these technologies in diverse biomedical applications is substantiated. Controlled molecular and cellular surface patterning is a costly and time-consuming process. Methods for keeping multiple separate experimental conditions on a patterned area are, therefore, needed to amplify the amount of biological information that can be retrieved from a patterned surface area. We describe, in three examples of biomedical applications, how this can be achieved in an open microfluidic system, by hydrodynamically guiding sample fluid over biological molecules and living cells immobilized on a surface. Results A microfluidic format of a standard assay for cell-membrane integrity showed a fast and dose-dependent toxicity of saponin on mammalian cells. A model of the interactions of human mononuclear leukocytes and endothelial cells was established. By contrast to static adhesion assays, cell-cell adhesion in this dynamic model depended on cytokine-mediated activation of both endothelial and blood cells. The microfluidic system allowed the use of unprocessed blood as sample material, and a specific and fast immunoassay for measuring the concentration of C-reactive protein in whole blood was demonstrated. Conclusion The use of hydrodynamic guiding made multiple and dynamic experimental conditions on a small surface area possible. The ability to change the direction of flow and produce two-dimensional grids can increase the number of reactions per surface area even further. The described microfluidic system is widely applicable, and can take advantage of surfaces produced by current and future techniques for patterning in the micro- and nanometer scale. PMID:12875662

  19. A fast prototyping process for fabrication of microfluidic systems on soda-lime glass

    NASA Astrophysics Data System (ADS)

    Lin, Che-Hsin; Lee, Gwo-Bin; Lin, Yen-Heng; Chang, Guan-Liang

    2001-11-01

    This paper describes a fast, low-cost but reliable process for the fabrication of microfluidic systems on soda-lime glass substrates. Instead of using an expensive metal or polisilicon/nitride layer as an etch mask, a thin layer of AZ 4620 positive photoresist (PR) is used for buffered oxide etching (BOE) of soda-lime glass. A novel two-step baking process prolongs the survival time of the PR mask in the etchant, which avoids serious peeling problems of the PR. A new process to remove precipitated particles generated during the etching process is also reported in which the glass substrate is dipped into a 1 M hydrochloride solution. A microfluidic channel with a depth of 35.95±0.39 µm is formed after 40 min BOE in an ultrasonic bath. The resulting channel has a smooth profile with a surface roughness of less than 45.95±7.96 Å. Glass chips with microfluidic channels are then bonded at 580 °C for 20 min to seal the channel while a slight pressure is applied. A new bonding process has been developed such that the whole process can be finished within 10 h. To our knowledge, this is the shortest processing time that has ever been reported. In the present study, an innovative microfluidic device, a `micro flow-through sampling chip', has been demonstrated using the fabrication method. Successful sampling and separation of Cy5-labelled bovine serum albumin (BSA) and anti-BSA has been achieved. This simple fabrication process is suitable for fast prototyping and mass production of microfluidic systems.

  20. Research on microfluidic chip and imaging system used to measure Ca2+ in cell

    NASA Astrophysics Data System (ADS)

    Zhou, Wei; Zhang, Sixiang; Ran, Dugang; Liu, Bao

    2009-07-01

    The microfluidic fluorescence detecting system which is used to measure the concentration of Ca2+ had been designed. On the microfluidic chip we designed, cell-dyeing, cell fostering, reagent injecting and other operations can be completed. The monochromatic light came from optical monochromator which can emit continuous spectrum was used to excitated the fluorescent probe in the cell, then the fluorescence signal and image were sampled by the PMT and CCD, at last the data was processed and the content of Ca2+ in the cell was figured out by using the fluorescence ratio method. Meanwhile, by using the system, the dynamic curve of [Ca2+]1 in cell was given after being stimulated by high K+. The precise result verifies that the system is stable and credible and it meets the requirement of detecting [Ca2+]i in live cells in the filed of physiology.

  1. A microfluidic two-pump system inspired by liquid feeding in mosquitoes

    NASA Astrophysics Data System (ADS)

    Marino, Andrew; Goad, Angela; Stremler, Mark; Socha, John; Jung, Sunghwan

    Mosquitoes feed on nectar and blood using a two-pump system in the head-a smaller cibarial pump in line with a larger a pharyngeal pump, with a valve in between. To suck, mosquitoes transport the liquid (which may be a multi-component viscous fluid, blood) through a long micro-channel, the proboscis. In the engineering realm, microfluidic devices in biomedical applications, such as lab-on-a-chip technology, necessitate implementing a robust pump design to handle clogging and increase flow control compared to a single-pump system. In this talk, we introduce a microfluidic pump design inspired by the mosquito's two-pump system. The pumping performance (flow rate) in presence of impurities (air bubbles, soft clogs) is quantified as a function of phase difference and volume expansion of the pumps, and the elasticity of the valve.

  2. MEMS-based flow cytometry: microfluidics-based cell identification system by fluorescent imaging.

    PubMed

    Wu, W K; Liang, C K; Huang, J Z

    2004-01-01

    This study utilizes MEMS technology to realize a novel low-cost microfluidics-based biochip system for flow-type cell handling. Powered by vacuum pump, the microfluidic driving system enables cells to move in order one by one in the biochip by an effect of sheath flow prefocus. Then, cells are guided to a fluorescent inspection region where two detection tasks such as cell image identification and cell counting are conducted. Currently, the glass-based biochip has been manufactured and all the related devices have been well set up in our laboratory. With this proposed prototype system, typical results about cell separation of yeast cell and PC-3 cell are available and their separated images are also presented, respectively. PMID:17270801

  3. [Development of rotating perfusion bioreactor system and application for bone tissue engineering].

    PubMed

    Li, Xiang; Li, Dichen; Wang, Lin; Wang, Zhen; Lu, Bingheng

    2007-02-01

    A rotating perfusion bioreactor system has recently been developed in our laboratory to produce 3D dynamic culture condition, and the critical-sized scaffolds with interconnected microchennels were fabricated. Gas exchange occurs by semipermeable membrane covered on each side of bioreactor and gas-permeable peristaltic pump tube. Rotation and perfusion of culture media through large scaffolds enhance well mixing and mass transport of oxygen and nutrients in the bioreactor. Osteoblastic cells attached to microchennels are exposed to a low fluid flow-induced shear stress level. This bioreactor system overcomes several defects exited in static culture condition, improves the culture environment, facilitates osteoblast proliferation, differntiation, significant matrix production and mineralization, and the controllability of culture process is enhanced. Large scaffolds/osteoblast constructs were cultured in the bioreactor system for 14 days. Osteoblastic cells attached to microchannels of scaffolds were observed under scanning electron microscope (SEM). The results indicated that cells grew extensively in the microchennels of large scaffolds. PMID:17333894

  4. Small-angle optical deflection from collinear configuration for sensitive detection in microfluidic systems.

    PubMed

    Yang, Li; Li, Xiangtang; Li, Jing; Yuan, Hongyan; Zhao, Shulin; Xiao, Dan

    2012-07-01

    This paper describes a novel detection system based on small-angle optical deflection from the collinear configuration of a microfluidic chip. In this system, the incident light beam was focused on the microchannel through the edge of a lens, resulting in a small deflection angle that deviated 20° from the collinear configuration. The emitted fluorescence was collected through the center of the same lens and delivered to a photomultiplier tube in the vertical direction; the reflection light of the chip plate was kept away from the detector. In contrast to traditional confocal and nonconfocal laser-induced fluorescence detection systems, background levels resulting from scattered excitation light, reflection and refraction from the microchip was significantly eliminated. Significant enhancement of the signal-to-noise ratio was obtained by shaping a laser beam that combined an attenuator with a spectral filter to optimize laser power and the dimensions of the laser beam. FITC and FITC-labeled amino acid were used as model analytes to demonstrate the performance sensitivity, separation efficiency, and reproducibility of this detection system by using a hybrid polydimethylsiloxane/glass microfluidic device. The limit of detection of FITC was estimated to be 2 pM (0.55 zmol) (S/N = 3). Furthermore, the single cell analysis for the determination of intracellular glutathione in a single 3T3 mouse fibroblast cell was demonstrated. The results suggest that the proposed optical arrangements will be promising for development of sensitive, low-cost microfluidic systems. PMID:22806465

  5. Plug-n-play microfluidic systems from flexible assembly of glass-based flow-control modules.

    PubMed

    Meng, Zhi-Jun; Wang, Wei; Liang, Xuan; Zheng, Wei-Chao; Deng, Nan-Nan; Xie, Rui; Ju, Xiao-Jie; Liu, Zhuang; Chu, Liang-Yin

    2015-04-21

    In this study, we report on a simple and versatile plug-n-play microfluidic system that is fabricated from flexible assembly of glass-based flow-control modules for flexibly manipulating flows for versatile emulsion generation. The microfluidic system consists of three basic functional units: a flow-control module, a positioning groove, and a connection fastener. The flow-control module that is based on simple assembly of low-cost glass slides, coverslips, and glass capillaries provides excellent chemical resistance and optical properties, and easy wettability modification for flow manipulation. The flexible combination of the flow-control modules with 3D-printed positioning grooves and connection fasteners enables creation of versatile microfluidic systems for generating various higher-order multiple emulsions. The simple and reversible connection of the flow-control modules also allows easy disassembly of the microfluidic systems for further scale-up and functionalization. We demonstrate the scalability and controllability of flow manipulation by creating microfluidic systems from flexible assembly of flow-control modules for controllable generation of multiple emulsions from double emulsions to quadruple emulsions. Meanwhile, the flexible flow manipulation in the flow-control module provides advanced functions for improved control of the drop size, and for controllable generation of drops containing distinct components within multiple emulsions to extend the emulsion structure. Such modular microfluidic systems provide flexibility and versatility to flexibly manipulate micro-flows for enhanced and extended applications. PMID:25711675

  6. High-speed CMOS image sensor for high-throughput lensless microfluidic imaging system

    NASA Astrophysics Data System (ADS)

    Yan, Mei; Huang, Xiwei; Jia, Qixiang; Nadipalli, Revanth; Wang, Tongxi; Shang, Yang; Yu, Hao; Je, Minkyu; Yeo, Kiatseng

    2012-03-01

    The integration of CMOS image sensor and microfluidics becomes a promising technology for point-of-care (POC) diagnosis. However, commercial image sensors usually have limited speed and low-light sensitivity. One high-speed and high-sensitivity CMOS image sensor chip is introduced in this paper, targeted for high-throughput microfluidic imaging system. Firstly, high speed image sensor architecture is introduced with design of column-parallel single-slope analog-todigital converter (ADC) with digital correlated double sampling (CDS). The frame rate can be achieved to 2400 frames/second (fps) with resolution of 128×96 for high-throughput microfluidic imaging. Secondly, the designed system has superior low-light sensitivity, which is achieved by large pixel size (10μm×10μm, 56% fill factor). Pixel peak signalnoise- ratio (SNR) reaches to 50dB with 10dB improvement compared to the commercial pixel (2.2μm×2.2μm). The degradation of pixel resolution is compensated by super-resolution image processing algorithm. By reconstructing single image with multiple low-resolution frames, we can equivalently achieve 2μm resolution with physical 10μm pixel. Thirdly, the system-on-chip (SoC) integration results in a real-time controlled intelligent imaging system without expensive data storage and time-consuming computer analysis. This initial sensor prototype with timing-control makes it possible to develop high-throughput lensless microfluidic imaging system for POC diagnosis.

  7. Finger-powered microfluidic systems using multilayer soft lithography and injection molding processes.

    PubMed

    Iwai, Kosuke; Shih, Kuan Cheng; Lin, Xiao; Brubaker, Thomas A; Sochol, Ryan D; Lin, Liwei

    2014-10-01

    Point-of-care (POC) and disposable biomedical applications demand low-power microfluidic systems with pumping components that provide controlled pressure sources. Unfortunately, external pumps have hindered the implementation of such microfluidic systems due to limitations associated with portability and power requirements. Here, we propose and demonstrate a 'finger-powered' integrated pumping system as a modular element to provide pressure head for a variety of advanced microfluidic applications, including finger-powered on-chip microdroplet generation. By utilizing a human finger for the actuation force, electrical power sources that are typically needed to generate pressure head were obviated. Passive fluidic diodes were designed and implemented to enable distinct fluids from multiple inlet ports to be pumped using a single actuation source. Both multilayer soft lithography and injection molding processes were investigated for device fabrication and performance. Experimental results revealed that the pressure head generated from a human finger could be tuned based on the geometric characteristics of the pumping system, with a maximum observed pressure of 7.6 ± 0.1 kPa. In addition to the delivery of multiple, distinct fluids into microfluidic channels, we also employed the finger-powered pumping system to achieve the rapid formation of both water-in-oil droplets (106.9 ± 4.3 μm in diameter) and oil-in-water droplets (75.3 ± 12.6 μm in diameter) as well as the encapsulation of endothelial cells in droplets without using any external or electrical controllers. PMID:25102160

  8. An electric stimulation system for electrokinetic particle manipulation in microfluidic devices

    NASA Astrophysics Data System (ADS)

    Lopez-de la Fuente, M. S.; Moncada-Hernandez, H.; Perez-Gonzalez, V. H.; Lapizco-Encinas, B. H.; Martinez-Chapa, S. O.

    2013-03-01

    Microfluidic devices have grown significantly in the number of applications. Microfabrication techniques have evolved considerably; however, electric stimulation systems for microdevices have not advanced at the same pace. Electric stimulation of micro-fluidic devices is an important element in particle manipulation research. A flexible stimulation instrument is desired to perform configurable, repeatable, automated, and reliable experiments by allowing users to select the stimulation parameters. The instrument presented here is a configurable and programmable stimulation system for electrokinetic-driven microfluidic devices; it consists of a processor, a memory system, and a user interface to deliver several types of waveforms and stimulation patterns. It has been designed to be a flexible, highly configurable, low power instrument capable of delivering sine, triangle, and sawtooth waveforms with one single frequency or two superimposed frequencies ranging from 0.01 Hz to 40 kHz, and an output voltage of up to 30 Vpp. A specific stimulation pattern can be delivered over a single time period or as a sequence of different signals for different time periods. This stimulation system can be applied as a research tool where manipulation of particles suspended in liquid media is involved, such as biology, medicine, environment, embryology, and genetics. This system has the potential to lead to new schemes for laboratory procedures by allowing application specific and user defined electric stimulation. The development of this device is a step towards portable and programmable instrumentation for electric stimulation on electrokinetic-based microfluidic devices, which are meant to be integrated with lab-on-a-chip devices.

  9. Integrated microwave resonant device for dielectric analysis of microfluidic systems

    NASA Astrophysics Data System (ADS)

    Rowe, D. J.; Porch, A.; Barrow, D. A.; Allender, C. J.

    2011-08-01

    Herein we present a device for performing non-contact dielectric spectroscopy upon liquids in a microfluidic environment. The device is comprised of a compression-sealed polytetrafluoroethylene (PTFE) chip with an embedded coaxial resonator, which is overmoded for dielectric measurements at six discrete frequencies between 1 and 8 GHz. A novel capacitive coupling structure allows transmission measurements to be taken from one end of the resonator, and an optimised microchannel design maximises sensitivity and repeatability. The use of a PTFE substrate and a non-contact measurement gives excellent chemical and biological compatibility. A simple 'fingerprint' method for identifying solvents is demonstrated, whereby a sample is characterised by air-referenced changes in complex frequency. Complex permittivity values are also obtained via a perturbation theory-based inversion. A combination of experimental and simulated results is used to characterise the device behaviour, limits of operation and measurement uncertainty. The high stability of temporal measurements, coupled with the robustness of the design, make this device ideal for analytical chemistry and industrial process control.

  10. Lattice Boltzmann Simulation of Particle Laden Flows in Microfluidic Systems

    SciTech Connect

    Clague, D S; Weisgraber, T; Wheeler, E; Hon, G; Radford, J; Gascoyne, P; Smity, R; Liepmann, D; Meinhart, C; Santiago, J; Krulevitch, P

    2003-07-22

    The goal of this effort was to develop dynamic simulation tools to study and characterize particulate transport in Microfluidic devices. This includes the effects of external fields and near-field particle-particle, particle-surface interactions. The unique aspect of this effort is that we focused on the particles in suspension and rigorously accounted for all of the interactions that they experienced in solution. In contrast, other numerical methods within the program, finite element and finite volume approaches, typically treat the suspended species as non-interacting point particles. Later in the program, some of these approaches incorporated approximations to begin to account for particle-particle interactions. Through the programs (BioFlips and SIMBIOSYS), we developed collaborative relationships with device-oriented efforts. More specifically and at the request of the SIMBIOSYS program manager, we allowed our efforts/milestones to be more guided by the needs of our BioFlips colleagues; therefore, our efforts were focused on the needs of the MD Anderson Cancer Center (Peter Gascoyne), UCDavis (Rosemary Smith), and UC Berkeley (Dorian Liepmann). The first two collaborations involved the development of Dielectrophoresis analysis tools and the later involved the development of suspension and fluid modeling tools for microneedles.

  11. An electromagnetic microvalve for pneumatic control of microfluidic systems.

    PubMed

    Liu, Xuling; Li, Songjing

    2014-10-01

    An electromagnetic microvalve for pneumatic control of microfluidic devices has been designed, fabricated, and tested. The microvalve is composed of two parts: a miniature electromagnetic actuator and a valve body. The electromagnetic actuator consists mainly of a thin polydimethylsiloxane (PDMS)-based elastomer, which acts as the valve diaphragm. The diaphragm, used as a solid hydraulic medium, converts the large contact area of a valve core into a small contact area of valve head while maintaining a large stroking force. This microvalve remains closed because of a compressed mechanical spring force generated by the actuator. On the other hand, when a voltage is applied, the valve core moves up, relaxing the thin PDMS membrane, opening the microvalve. The fast open response (~17 ms) of the valve was achieved with a leak rate as low as 0.026 sccm at 200 KPa (N2) pressure. We tested the pertinent dynamic parameters such as flow rate in on/off mode, flow rate of duty cycles, and actuated frequencies in pulse width modulation (PWM) mode. Our method provides a simple, cheap, and small microvalve that avoids the bulky and expensive external pressure control solenoid manifold. This allows it to be easily integrated into portable and disposable devices. PMID:24742860

  12. A droplet microfluidic system for sequential generation of lipid bilayers and transmembrane electrical recordings.

    PubMed

    Czekalska, Magdalena A; Kaminski, Tomasz S; Jakiela, Slawomir; Tanuj Sapra, K; Bayley, Hagan; Garstecki, Piotr

    2015-01-21

    This paper demonstrates a microfluidic system that automates i) formation of a lipid bilayer at the interface between a pair of nanoliter-sized aqueous droplets in oil, ii) exchange of one droplet of the pair to form a new bilayer, and iii) current measurements on single proteins. A new microfluidic architecture is introduced - a set of traps designed to localize the droplets with respect to each other and with respect to the recording electrodes. The system allows for automated execution of experimental protocols by active control of the flow on chip with the use of simple external valves. Formation of stable artificial lipid bilayers, incorporation of α-hemolysin into the bilayers and electrical measurements of ionic transport through the protein pore are demonstrated. PMID:25412368

  13. Microfluidics-Based Lab-on-Chip Systems in DNA-Based Biosensing: An Overview

    PubMed Central

    Dutse, Sabo Wada; Yusof, Nor Azah

    2011-01-01

    Microfluidics-based lab-on-chip (LOC) systems are an active research area that is revolutionising high-throughput sequencing for the fast, sensitive and accurate detection of a variety of pathogens. LOCs also serve as portable diagnostic tools. The devices provide optimum control of nanolitre volumes of fluids and integrate various bioassay operations that allow the devices to rapidly sense pathogenic threat agents for environmental monitoring. LOC systems, such as microfluidic biochips, offer advantages compared to conventional identification procedures that are tedious, expensive and time consuming. This paper aims to provide a broad overview of the need for devices that are easy to operate, sensitive, fast, portable and sufficiently reliable to be used as complementary tools for the control of pathogenic agents that damage the environment. PMID:22163925

  14. Modular microfluidic cartridge-based universal diagnostic system for global health applications

    NASA Astrophysics Data System (ADS)

    Becker, Holger; Klemm, Richard; Dietze, William; White, Wallace; Hlawatsch, Nadine; Freyberg, Susanne; Moche, Christian; Dailey, Peter; Gärtner, Claudia

    2016-03-01

    Current microfluidics-enabled point-of-care diagnostic systems are typically designed specifically for one assay type, e.g. a molecular diagnostic assay for a single disease of a class of diseases. This approach often leads to high development cost and a significant training requirement for users of different instruments. We have developed an open platform diagnostic system which allows to run molecular, immunological and clinical assays on a single instrument platform with a standardized microfluidic cartridge architecture in an automated sample-in answer-out fashion. As examples, a molecular diagnostic assay for tuberculosis, an immunoassay for HIV p24 and a clinical chemistry assay for ALT liver function have been developed and results of their pre-clinical validation are presented.

  15. Hydrodynamic directional control of liquid metal droplets within a microfluidic flow focusing system

    NASA Astrophysics Data System (ADS)

    Gol, Berrak; Kurdzinski, Michael E.; Tovar-Lopez, Francisco J.; Petersen, Phred; Mitchell, Arnan; Khoshmanesh, Khashayar

    2016-04-01

    Here, we investigate the directional control of Galinstan liquid metal droplets when transferring from the high-viscosity glycerol core into the parallel low-viscosity NaOH sheath streams within a flow focusing microfluidic system. In the presence of sufficient flow mismatch between the sheath streams, the droplets are driven toward the higher velocity interface and cross the interface under the influence of surface tension gradient. A minimum flow mismatch of 125 μl/min is required to enable the continuous transfer of droplets toward the desired sheath stream. The response time of droplets, the time required to change the direction of droplet transfer, is governed by the response time of the syringe pump driven microfluidic system and is found to be 3.3 and 8.8 s when increasing and decreasing the flow rate of sheath stream, respectively.

  16. Laser ablated micropillar energy directors for ultrasonic welding of microfluidic systems

    NASA Astrophysics Data System (ADS)

    Esben Poulsen, Carl; Kistrup, Kasper; Korsgaard Andersen, Nis; Taboryski, Rafael; Fougt Hansen, Mikkel; Wolff, Anders

    2016-06-01

    We present a new type of energy director (ED) for ultrasonic welding of microfluidic systems. These micropillar EDs are based on the replication of cone like protrusion structures introduced using a pico-second laser and may therefore be added to any mould surface accessible to a pico-second laser beam. The technology is demonstrated on an injection moulded microfluidic device featuring high-aspect ratio (h  ×  w  =  2000 μm  ×  550 μm) and free-standing channel walls, where bonding is achieved with no detectable channel deformation. The bonding strength is similar to conventional EDs and the fabricated system can withstand pressures of over 9.5 bar.

  17. Design of liquid lens based optical system for microfluidic chip detection

    NASA Astrophysics Data System (ADS)

    Niu, Lijun; Zhou, Ya; Hu, Xiaoming; Zhou, Chang

    2015-08-01

    The precision of manufacturing and installing together with the flexibility is a serious challenge for laser induced fluorescent detector (LIFD) of microfluidic chip. In this paper, a focus tunable liquid lens based on liquid zoom system for LIFD with automatic adjustment is proposed. With the help of liquid zoom lens whose surface curvature can be varied continuously by current, the system can achieve a continuous zoom. Instead of using the traditional mechanical axial displacement scanning motion mechanism, the proposed zoom system can implement axial displacement scan by means of the well-designed autofocus feedback current control function. The simulation results show that the focal length variation range of the designed optical system is 4.87mm~ 8.40mm, which is also the axial scanning displacement range. The size of scanning spot is around 15μm when a 473nm wavelength laser is used, which can meet the demand of microfluidic chip detection. With this design, the required precision of the LIFD could be reduced significantly as well as costs. Moreover, it also makes the detection of microfluidic chip qualified to adapt to different size of detecting channel.

  18. Microfluidic system for high-throughput immunoglobulin-E analysis from clinical serum samples.

    PubMed

    Zheng, Lulu; Fu, Yongfeng; Jiang, Xiran; Man, Suqin; Ran, Wei; Feng, Meng; Liu, Sixiu; Cheng, Xunjia; Sui, Guodong

    2015-10-01

    Rapid and high-throughput analytical techniques for IgE that requires a small serum amount are very important, especially for pediatric patients. In these patients, blood is collected from veins, which is painful compared to fingertip blood collection. Herein, a novel microfluidic system capable of high-throughput parallel analyses of allergen-specific IgE from small amounts of patient serum was successfully developed. A six-plex immunoassay was constructed within a microfluidic chip, and the entire system was validated using samples from clinical patients. Major antigens from house dust mite (Dermatophagoides farinae and Blomia tropicalis), cat (Felis domesticus), fungus (Cladosporium herbarum), ragweed (Humulus japonicas), and tree pollen (Platanus acerifolia) were used as analysis targets. Sample consumption decreased to <0.05 µL compared with the 480µL serum consumption by fluoroenzyme immunoassay (UniCAP system Pharmacia Diagnostics AB, Uppsala, Sweden), the 50 µL serum consumption by enzyme-linked immune sorbent assay (ELISA), or the 1.5 µL serum consumption by conventional protein chip analysis. Analysis duration, reagent cost, and total cost for each measurement were also considerably decreased. The assay showed good accuracy and sensitivity toward the clinical samples. A significant correlation of allergen-specific IgE levels was found among the microfluidic assay, UniCAP system, and ELISA. PMID:26078132

  19. DEM microfabrication technique and its applications in bioscience and microfluidic systems

    NASA Astrophysics Data System (ADS)

    Chen, Di; Yang, Fan; Tang, Min; Li, Yigui; Zhu, Jun; Zhang, Dacheng

    2001-10-01

    A new LIGA-like microfabrication technique was developed by present authors. DEM (deepetching, electroforming and microreplication) is the abbreviation of three main process steps in this new microfabrication technique. In contrast to LIGA technique, DEM technique has the advantages of lower cost and shorter process period. Microfluidic systems like plastic capillary electrophoresis chips, micro flowmeters and three-dimensional DNA chips were developed using DEM technique. DEM technique offers a new way for fabrication of MEMS and MOEMS components.

  20. Microfluidic fuel cell systems with embedded materials and structures and method thereof

    DOEpatents

    Morse, Jeffrey D.; Rose, Klint A; Maghribi, Mariam; Benett, William; Krulevitch, Peter; Hamilton, Julie; Graff, Robert T.; Jankowski, Alan

    2005-07-26

    Described herein is a process for fabricating microfluidic systems with embedded components in which micron-scale features are molded into the polymeric material polydimethylsiloxane (PDMS). Micromachining is used to create a mold master and the liquid precursors for PDMS are poured over the mold and allowed to cure. The PDMS is then removed form the mold and bonded to another material such as PDMS, glass, or silicon after a simple surface preparation step to form sealed microchannels.

  1. The effect of nifedipine on myocardial perfusion and metabolism in systemic sclerosis. A positron emission tomographic study

    SciTech Connect

    Duboc, D.; Kahan, A.; Maziere, B.; Loc'h, C.; Crouzel, C.; Menkes, C.J.; Amor, B.; Strauch, G.; Guerin, F.; Syrota, A. )

    1991-02-01

    We assessed the effect of nifedipine on myocardial perfusion and metabolism in 9 patients with systemic sclerosis, using positron emission tomography with a perfusion tracer (potassium-38) and a metabolic tracer (18F-fluorodeoxyglucose (18FDG)). Nifedipine, 20 mg 3 times daily for 1 week, induced a significant increase in 38K myocardial uptake, a significant decrease in 18FDG myocardial uptake, and a significant increase in the myocardial 38K: 18FDG ratio. These results indicate that the increase in myocardial perfusion is associated with modifications in myocardial energy metabolism, which probably result from a beneficial anti-ischemic effect of nifedipine in patients with systemic sclerosis.

  2. Integrated Cantilever-Based Flow Sensors with Tunable Sensitivity for In-Line Monitoring of Flow Fluctuations in Microfluidic Systems

    PubMed Central

    Noeth, Nadine; Keller, Stephan Sylvest; Boisen, Anja

    2014-01-01

    For devices such as bio-/chemical sensors in microfluidic systems, flow fluctuations result in noise in the sensor output. Here, we demonstrate in-line monitoring of flow fluctuations with a cantilever-like sensor integrated in a microfluidic channel. The cantilevers are fabricated in different materials (SU-8 and SiN) and with different thicknesses. The integration of arrays of holes with different hole size and number of holes allows the modification of device sensitivity, theoretical detection limit and measurement range. For an average flow in the microliter range, the cantilever deflection is directly proportional to the flow rate fluctuations in the microfluidic channel. The SiN cantilevers show a detection limit below 1 nL/min and the thinnest SU-8 cantilevers a detection limit below 5 nL/min. Finally, the sensor is applied for in-line monitoring of flow fluctuations generated by external pumps connected to the microfluidic system. PMID:24366179

  3. Micro-fluidic interconnect

    DOEpatents

    Okandan, Murat; Galambos, Paul C.; Benavides, Gilbert L.; Hetherington, Dale L.

    2006-02-28

    An apparatus for simultaneously aligning and interconnecting microfluidic ports is presented. Such interconnections are required to utilize microfluidic devices fabricated in Micro-Electromechanical-Systems (MEMS) technologies, that have multiple fluidic access ports (e.g. 100 micron diameter) within a small footprint, (e.g. 3 mm.times.6 mm). Fanout of the small ports of a microfluidic device to a larger diameter (e.g. 500 microns) facilitates packaging and interconnection of the microfluidic device to printed wiring boards, electronics packages, fluidic manifolds etc.

  4. Microfluidics-Based in Vivo Mimetic Systems for the Study of Cellular Biology

    PubMed Central

    2015-01-01

    Conspectus The human body is a complex network of molecules, organelles, cells, tissues, and organs: an uncountable number of interactions and transformations interconnect all the system’s components. In addition to these biochemical components, biophysical components, such as pressure, flow, and morphology, and the location of all of these interactions play an important role in the human body. Technical difficulties have frequently limited researchers from observing cellular biology as it occurs within the human body, but some state-of-the-art analytical techniques have revealed distinct cellular behaviors that occur only in the context of the interactions. These types of findings have inspired bioanalytical chemists to provide new tools to better understand these cellular behaviors and interactions. What blocks us from understanding critical biological interactions in the human body? Conventional approaches are often too naïve to provide realistic data and in vivo whole animal studies give complex results that may or may not be relevant for humans. Microfluidics offers an opportunity to bridge these two extremes: while these studies will not model the complexity of the in vivo human system, they can control the complexity so researchers can examine critical factors of interest carefully and quantitatively. In addition, the use of human cells, such as cells isolated from donated blood, captures human-relevant data and limits the use of animals in research. In addition, researchers can adapt these systems easily and cost-effectively to a variety of high-end signal transduction mechanisms, facilitating high-throughput studies that are also spatially, temporally, or chemically resolved. These strengths should allow microfluidic platforms to reveal critical parameters in the human body and provide insights that will help with the translation of pharmacological advances to clinical trials. In this Account, we describe selected microfluidic innovations within the

  5. Toward complete miniaturisation of flow injection analysis systems: microfluidic enhancement of chemiluminescent detection.

    PubMed

    Gracioso Martins, Ana M; Glass, Nick R; Harrison, Sally; Rezk, Amgad R; Porter, Nichola A; Carpenter, Peter D; Du Plessis, Johan; Friend, James R; Yeo, Leslie Y

    2014-11-01

    Conventional flow injection systems for aquatic environmental analysis typically comprise large laboratory benchscale equipment, which place considerable constraints for portable field use. Here, we demonstrate the use of an integrated acoustically driven microfluidic mixing scheme to enhance detection of a chemiluminescent species tris(2,2'-bipyridyl)dichlororuthenium(II) hexahydrate-a common chemiluminescent reagent widely used for the analysis of a wide range of compounds such as illicit drugs, pharmaceuticals, and pesticides-such that rapid in-line quantification can be carried out with sufficient on-chip sensitivity. Specifically, we employ surface acoustic waves (SAWs) to drive intense chaotic streaming within a 100 μL chamber cast in polydimethoxylsiloxane (PDMS) atop a microfluidic chip consisting of a single crystal piezoelectric material. By optimizing the power, duration, and orientation of the SAW input, we show that the mixing intensity of the sample and reagent fed into the chamber can be increased by one to two orders of magnitude, leading to a similar enhancement in the detection sensitivity of the chemiluminescent species and thus achieving a theoretical limit of detection of 0.02 ppb (0.2 nM) of l-proline-a decade improvement over the industry gold-standard and two orders of magnitude more sensitive than that achievable with conventional systems-simply using a portable photodetector and without requiring sample preconcentration. This on-chip microfluidic mixing strategy, together with the integrated miniature photodetector and the possibility for chip-scale microfluidic actuation, then alludes to the attractive possibility of a completely miniaturized platform for portable field-use microanalytical systems. PMID:25275830

  6. Modeling and simulation of DNA flow in a microfluidic-based pathogen detection system

    SciTech Connect

    Trebotich, D; Miller, G H

    2005-01-31

    We present simulation results from a new computational model of DNA flow in microfluidic devices. This work is important because computational models are needed to design miniaturized biomedical devices that are becoming the state-of-the-art in many significant applications including pathogen detection as well as continuous monitoring and drug delivery. Currently advanced algorithms in design tools are non-existent but necessary to understand the complex fluid and polymer dynamics involved in biological flow at small scales. Our model is based on a fully coupled fluid-particle numerical algorithm with both stochastic and deterministic components in a bead-rod polymer representation. We have applied this work to DNA extraction configurations in a microfluidic PCR chamber used in a pathogen detection system. We demonstrate our method on the test problem of flow of a single DNA molecule in a 2D packed array microchannel. We are also investigating mechanisms for molecular ''sticking'' using short range forces.

  7. A fully integrated microfluidic genetic analysis system with sample-in-answer-out capability.

    PubMed

    Easley, Christopher J; Karlinsey, James M; Bienvenue, Joan M; Legendre, Lindsay A; Roper, Michael G; Feldman, Sanford H; Hughes, Molly A; Hewlett, Erik L; Merkel, Tod J; Ferrance, Jerome P; Landers, James P

    2006-12-19

    We describe a microfluidic genetic analysis system that represents a previously undescribed integrated microfluidic device capable of accepting whole blood as a crude biological sample with the endpoint generation of a genetic profile. Upon loading the sample, the glass microfluidic genetic analysis system device carries out on-chip DNA purification and PCR-based amplification, followed by separation and detection in a manner that allows for microliter samples to be screened for infectious pathogens with sample-in-answer-out results in < 30 min. A single syringe pump delivers sample/reagents to the chip for nucleic acid purification from a biological sample. Elastomeric membrane valving isolates each distinct functional region of the device and, together with resistive flow, directs purified DNA and PCR reagents from the extraction domain into a 550-nl chamber for rapid target sequence PCR amplification. Repeated pressure-based injections of nanoliter aliquots of amplicon (along with the DNA sizing standard) allow electrophoretic separation and detection to provide DNA fragment size information. The presence of Bacillus anthracis (anthrax) in 750 nl of whole blood from living asymptomatic infected mice and of Bordetella pertussis in 1 microl of nasal aspirate from a patient suspected of having whooping cough are confirmed by the resultant genetic profile. PMID:17159153

  8. A fluorescence-based centrifugal microfluidic system for parallel detection of multiple allergens

    NASA Astrophysics Data System (ADS)

    Chen, Q. L.; Ho, H. P.; Cheung, K. L.; Kong, S. K.; Suen, Y. K.; Kwan, Y. W.; Li, W. J.; Wong, C. K.

    2010-02-01

    This paper reports a robust polymer based centrifugal microfluidic analysis system that can provide parallel detection of multiple allergens in vitro. Many commercial food products (milk, bean, pollen, etc.) may introduce allergy to people. A low-cost device for rapid detection of allergens is highly desirable. With this as the objective, we have studied the feasibility of using a rotating disk device incorporating centrifugal microfluidics for performing actuationfree and multi-analyte detection of different allergen species with minimum sample usage and fast response time. Degranulation in basophils or mast cells is an indicator to demonstrate allergic reaction. In this connection, we used acridine orange (AO) to demonstrate degranulation in KU812 human basophils. It was found that the AO was released from granules when cells were stimulated by ionomycin, thus signifying the release of histamine which accounts for allergy symptoms [1-2]. Within this rotating optical platform, major microfluidic components including sample reservoirs, reaction chambers, microchannel and flow-control compartments are integrated into a single bio-compatible polydimethylsiloxane (PDMS) substrate. The flow sequence and reaction time can be controlled precisely. Sequentially through varying the spinning speed, the disk may perform a variety of steps on sample loading, reaction and detection. Our work demonstrates the feasibility of using centrifugation as a possible immunoassay system in the future.

  9. Microprocessor-based integration of microfluidic control for the implementation of automated sensor monitoring and multithreaded optimization algorithms.

    PubMed

    Ezra, Elishai; Maor, Idan; Bavli, Danny; Shalom, Itai; Levy, Gahl; Prill, Sebastian; Jaeger, Magnus S; Nahmias, Yaakov

    2015-08-01

    Microfluidic applications range from combinatorial synthesis to high throughput screening, with platforms integrating analog perfusion components, digitally controlled micro-valves and a range of sensors that demand a variety of communication protocols. Currently, discrete control units are used to regulate and monitor each component, resulting in scattered control interfaces that limit data integration and synchronization. Here, we present a microprocessor-based control unit, utilizing the MS Gadgeteer open framework that integrates all aspects of microfluidics through a high-current electronic circuit that supports and synchronizes digital and analog signals for perfusion components, pressure elements, and arbitrary sensor communication protocols using a plug-and-play interface. The control unit supports an integrated touch screen and TCP/IP interface that provides local and remote control of flow and data acquisition. To establish the ability of our control unit to integrate and synchronize complex microfluidic circuits we developed an equi-pressure combinatorial mixer. We demonstrate the generation of complex perfusion sequences, allowing the automated sampling, washing, and calibrating of an electrochemical lactate sensor continuously monitoring hepatocyte viability following exposure to the pesticide rotenone. Importantly, integration of an optical sensor allowed us to implement automated optimization protocols that require different computational challenges including: prioritized data structures in a genetic algorithm, distributed computational efforts in multiple-hill climbing searches and real-time realization of probabilistic models in simulated annealing. Our system offers a comprehensive solution for establishing optimization protocols and perfusion sequences in complex microfluidic circuits. PMID:26227212

  10. Fabrication and integration of microprism mirrors for high-speed three-dimensional measurement in inertial microfluidic system

    NASA Astrophysics Data System (ADS)

    Koh, Joonyoung; Kim, Jihye; Shin, Jung H.; Lee, Wonhee

    2014-09-01

    Inertial microfluidics utilizes fluid inertia from high flow velocity to manipulate particles and fluids in 3D. Acquiring a 3D information of particle positions and complex flow patterns within microfluidic devices requires 3D imaging techniques such as confocal microscopy, which are often expensive and slow. Here, we report on a prism-mirror-embedded microfluidic device that allows simultaneous imaging of the top and side view of the microchannel for a high-speed, low-cost 3D imaging. The microprism mirrors are fabricated and integrated into a microfluidic system using conventional microfabrication techniques including wet etch and soft lithography. This inexpensive high quality prism mirror provides a highly reflective, smooth mirror surface with precise 45° reflection angle, enabling 3D measurement of inertial migration of microparticles in a rectangular channel at speeds in excess of 10 000 frame/s.

  11. Note: A micro-perfusion system for use during real-time physiological studies under high pressure

    NASA Astrophysics Data System (ADS)

    Maltas, Jeff; Long, Zac; Huff, Alison; Maloney, Ryan; Ryan, Jordan; Urayama, Paul

    2014-10-01

    We construct a micro-perfusion system using piston screw pump generators for use during real-time, high-pressure physiological studies. Perfusion is achieved using two generators, with one generator being compressed while the other is retracted, thus maintaining pressurization while producing fluid flow. We demonstrate control over perfusion rates in the 10-μl/s range and the ability to change between fluid reservoirs at up to 50 MPa. We validate the screw-pump approach by monitoring the cyanide-induced response of UV-excited autofluorescence from Saccharomyces cerevisiae under pressurization.

  12. Versatile minimized system - a step towards safe perfusion.

    PubMed

    Ganushchak, Y M; Körver, E P J; Yamamoto, Y; Weerwind, P W

    2016-05-01

    A growing body of evidence indicates the superiority of minimized cardiopulmonary bypass (CPB) systems compared to conventional systems in terms of inflammatory reactions and transfusion requirements. Evident benefits of minimized CPB systems, however, do not come without consequences. Kinetic-assisted drainage, as used in these circuits, can result in severe fluctuations of venous line pressures and, consequently, fluctuation of the blood flow delivered to the patient. Furthermore, subatmospheric venous line pressures can cause gaseous microemboli. Another limitation is the absence of cardiotomy suction, which can lead to excessive blood loss via a cell saver. The most serious limitation of minimized circuits is that these circuits are very constrained in the case of complications or changing of the surgery plan. We developed a versatile minimized system (VMS) with a priming volume of about 600 ml. A compliance chamber in the venous line decreases peaks of pressure fluctuations. This chamber also acts as a bubble trap. Additionally, the open venous reservoir is connected parallel to the venous line and excluded from the circulation during an uncomplicated CPB. This reservoir can be included in the circulation via a roller pump and be used as a cardiotomy reservoir. The amount and rate of returned blood in the circulation is regulated by a movable level detector. Further, the circuit can easily be converted to an open system with vacuum-assisted venous drainage in the case of unexpected complications. The VMS combines the benefits of minimized circuits with the versatility and safety of a conventional CPB system. Perfusionists familiar with this system can secure an adequate and timely response at expected and unexpected intraoperative complications. PMID:26354746

  13. Nanomaterial based detection and degradation of biological and chemical contaminants in a microfluidic system

    NASA Astrophysics Data System (ADS)

    Jayamohan, Harikrishnan

    Monitoring and remediation of environmental contaminants (biological and chemical) form the crux of global water resource management. There is an extant need to develop point-of-use, low-power, low-cost tools that can address this problem effectively with minimal environmental impact. Nanotechnology and microfluidics have made enormous advances during the past decade in the area of biosensing and environmental remediation. The "marriage" of these two technologies can effectively address some of the above-mentioned needs. In this dissertation, nanomaterials were used in conjunction with microfluidic techniques to detect and degrade biological and chemical pollutants. In the first project, a point-of-use sensor was developed for detection of trichloroethylene (TCE) from water. A self-organizing nanotubular titanium dioxide (TNA) synthesized by electrochemical anodization and functionalized with photocatalytically deposited platinum (Pt/TNA) was applied to the detection. The morphology and crystallinity of the Pt/TNA sensor was characterized using field emission scanning electron microscope, energy dis- persive x-ray spectroscopy, and X-ray diffraction. The sensor could detect TCE in the concentrations ranging from 10 to 1000 ppm. The room-temperature operation capability of the sensor makes it less power intensive and can potentially be incorporated into a field-based sensor. In the second part, TNA synthesized on a foil was incorporated into a flow-based microfluidic format and applied to degradation of a model pollutant, methylene blue. The system was demonstrated to have enhanced photocatalytic performance at higher flow rates (50-200 muL/min) over the same microfluidic format with TiO2 nanoparticulate (commercial P25) catalyst. The microfluidic format with TNA catalyst was able to achieve 82% fractional conversion of 18 mM methylene blue in comparison to 55% in the case of the TiO2 nanoparticulate layer at a flow rate of 200 L/min. The microfluidic device was

  14. Uniform mixing in paper-based microfluidic systems using surface acoustic waves.

    PubMed

    Rezk, Amgad R; Qi, Aisha; Friend, James R; Li, Wai Ho; Yeo, Leslie Y

    2012-02-21

    Paper-based microfluidics has recently received considerable interest due to their ease and low cost, making them extremely attractive as point-of-care diagnostic devices. The incorporation of basic fluid actuation and manipulation schemes on paper substrates, however, afford the possibility to extend the functionality of this simple technology to a much wider range of typical lab-on-a-chip operations, given its considerable advantages in terms of cost, size and integrability over conventional microfluidic substrates. We present a convective actuation mechanism in a simple paper-based microfluidic device using surface acoustic waves to drive mixing. Employing a Y-channel structure patterned onto paper, the mixing induced by the 30 MHz acoustic waves is shown to be consistent and rapid, overcoming several limitations associated with its capillary-driven passive mixing counterpart wherein irreproducibilities and nonuniformities are often encountered in the mixing along the channel--capillary-driven passive mixing offers only poor control, is strongly dependent on the paper's texture and fibre alignment, and permits backflow, all due to the scale of the fibres being significant in comparison to the length scales of the features in a microfluidic system. Using a novel hue-based colourimetric technique, the mixing speed and efficiency is compared between the two methods, and used to assess the effects of changing the input power, channel tortuousity and fibre/flow alignment for the acoustically-driven mixing. The hue-based technique offers several advantages over grayscale pixel intensity analysis techniques in facilitating quantification without limitations on the colour contrast of the samples, and can be used, for example, for quantification in on-chip immunochromatographic assays. PMID:22193520

  15. Droplet-based microfluidic system to form and separate multicellular spheroids using magnetic nanoparticles.

    PubMed

    Yoon, Sungjun; Kim, Jeong Ah; Lee, Seung Hwan; Kim, Minsoo; Park, Tai Hyun

    2013-04-21

    The importance of creating a three-dimensional (3-D) multicellular spheroid has recently been gaining attention due to the limitations of monolayer cell culture to precisely mimic in vivo structure and cellular interactions. Due to this emerging interest, researchers have utilized new tools, such as microfluidic devices, that allow high-throughput and precise size control to produce multicellular spheroids. We have developed a droplet-based microfluidic system that can encapsulate both cells and magnetic nanoparticles within alginate beads to mimic the function of a multicellular tumor spheroid. Cells were entrapped within the alginate beads along with magnetic nanoparticles, and the beads of a relatively uniform size (diameters of 85% of the beads were 170-190 μm) were formed in the oil phase. These beads were passed through parallel streamlines of oil and culture medium, where the beads were magnetically transferred into the medium phase from the oil phase using an external magnetic force. This microfluidic chip eliminates additional steps for collecting the spheroids from the oil phase and transferring them to culture medium. Ultimately, the overall spheroid formation process can be achieved on a single microchip. PMID:23426090

  16. Novel localized heating technique on centrifugal microfluidic disc with wireless temperature monitoring system.

    PubMed

    Joseph, Karunan; Ibrahim, Fatimah; Cho, Jongman

    2015-01-01

    Recent advances in the field of centrifugal microfluidic disc suggest the need for electrical interface in the disc to perform active biomedical assays. In this paper, we have demonstrated an active application powered by the energy harvested from the rotation of the centrifugal microfluidic disc. A novel integration of power harvester disc onto centrifugal microfluidic disc to perform localized heating technique is the main idea of our paper. The power harvester disc utilizing electromagnetic induction mechanism generates electrical energy from the rotation of the disc. This contributes to the heat generation by the embedded heater on the localized heating disc. The main characteristic observed in our experiment is the heating pattern in relative to the rotation of the disc. The heating pattern is monitored wirelessly with a digital temperature sensing system also embedded on the disc. Maximum temperature achieved is 82 °C at rotational speed of 2000 RPM. The technique proves to be effective for continuous heating without the need to stop the centrifugal motion of the disc. PMID:26736977

  17. A pneumatic pressure-driven multi-throughput microfluidic circulation culture system.

    PubMed

    Satoh, T; Narazaki, G; Sugita, R; Kobayashi, H; Sugiura, S; Kanamori, T

    2016-06-21

    Here, we report a pneumatic pressure-driven microfluidic device capable of multi-throughput medium circulation culture. The circulation culture system has the following advantages for application in drug discovery: (i) simultaneous operation of multiple circulation units, (ii) use of a small amount of circulating medium (3.5 mL), (iii) pipette-friendly liquid handling, and (iv) a detachable interface with pneumatic pressure lines via sterile air-vent filters. The microfluidic device contains three independent circulation culture units, in which human umbilical vein endothelial cells (HUVECs) were cultured under physiological shear stress induced by circulation of the medium. Circulation of the medium in the three culture units was generated by programmed sequentially applied pressure from two pressure-control lines. HUVECs cultured in the microfluidic device were aligned under a one-way circulating flow with a shear stress of 10 dyn cm(-2); they exhibited a randomly ordered alignment under no shear stress and under reciprocating flow with a shear stress of 10 dyn cm(-2). We also observed 2.8- to 4.9-fold increases in expression of the mRNAs of endothelial nitric oxide synthase and thrombomodulin under one-way circulating flow with a shear stress of 10 dyn cm(-2) compared with conditions of no shear stress or reciprocating flow. PMID:27229626

  18. Microfluidic bioassay system based on microarrays of hydrogel sensing elements entrapping quantum dot-enzyme conjugates.

    PubMed

    Jang, Eunji; Kim, Sinyoung; Koh, Won-Gun

    2012-01-15

    This paper presents a simple method to fabricate a microfluidic biosensor that is able to detect substrates for H(2)O(2)-generating oxidase. The biosensor consists of three components (quantum dot-enzyme conjugates, hydrogel microstructures, and a set of microchannels) that were hierarchically integrated into a microfluidic device. The quantum dot (QD)-enzyme conjugates were entrapped within the poly(ethylene glycol) (PEG)-based hydrogel microstructures that were fabricated within the microchannels by a photopatterning process. Glucose oxidase (GOX) and alcohol oxidase (AOX) were chosen as the model oxidase enzymes, conjugated to carboxyl-terminated CdSe/ZnS QDs, and entrapped within the hydrogel microstructures, which resulted in a fluorescent hydrogel microarray that was responsive to glucose or alcohol. The hydrogel-entrapped GOX and AOX were able to perform enzyme-catalyzed oxidation of glucose and alcohol, respectively, to produce H(2)O(2), which subsequently quenched the fluorescence of the conjugated QDs. The fluorescence intensity of the hydrogel microstructures decreased as the glucose and alcohol concentrations increased, and the detection limits of this system were found to be 50 μM of glucose and 70 μM of alcohol. Because each microchannel was able to carry out different assays independently, the simultaneous detection of glucose and alcohol was possible using our novel microfluidic device composed of multiple microchannels. PMID:22177543

  19. Comparative Single-Cell Analysis of Different E. coli Expression Systems during Microfluidic Cultivation

    PubMed Central

    Hilgers, Fabienne; Loeschcke, Anita; Jaeger, Karl-Erich; Kohlheyer, Dietrich; Drepper, Thomas

    2016-01-01

    Recombinant protein production is mostly realized with large-scale cultivations and monitored at the level of the entire population. Detailed knowledge of cell-to-cell variations with respect to cellular growth and product formation is limited, even though phenotypic heterogeneity may distinctly hamper overall production yields, especially for toxic or difficult-to-express proteins. Unraveling phenotypic heterogeneity is thus a key aspect in understanding and optimizing recombinant protein production in biotechnology and synthetic biology. Here, microfluidic single-cell analysis serves as the method of choice to investigate and unmask population heterogeneities in a dynamic and spatiotemporal fashion. In this study, we report on comparative microfluidic single-cell analyses of commonly used E. coli expression systems to uncover system-inherent specifications in the synthetic M9CA growth medium. To this end, the PT7lac/LacI, the PBAD/AraC and the Pm/XylS system were systematically analyzed in order to gain detailed insights into variations of growth behavior and expression phenotypes and thus to uncover individual strengths and deficiencies at the single-cell level. Specifically, we evaluated the impact of different system-specific inducers, inducer concentrations as well as genetic modifications that affect inducer-uptake and regulation of target gene expression on responsiveness and phenotypic heterogeneity. Interestingly, the most frequently applied expression system based on E. coli strain BL21(DE3) clearly fell behind with respect to expression homogeneity and robustness of growth. Moreover, both the choice of inducer and the presence of inducer uptake systems proved crucial for phenotypic heterogeneity. Conclusively, microfluidic evaluation of different inducible E. coli expression systems and setups identified the modified lacY-deficient PT7lac/LacI as well as the Pm/XylS system with conventional m-toluic acid induction as key players for precise and robust

  20. Comparative Single-Cell Analysis of Different E. coli Expression Systems during Microfluidic Cultivation.

    PubMed

    Binder, Dennis; Probst, Christopher; Grünberger, Alexander; Hilgers, Fabienne; Loeschcke, Anita; Jaeger, Karl-Erich; Kohlheyer, Dietrich; Drepper, Thomas

    2016-01-01

    Recombinant protein production is mostly realized with large-scale cultivations and monitored at the level of the entire population. Detailed knowledge of cell-to-cell variations with respect to cellular growth and product formation is limited, even though phenotypic heterogeneity may distinctly hamper overall production yields, especially for toxic or difficult-to-express proteins. Unraveling phenotypic heterogeneity is thus a key aspect in understanding and optimizing recombinant protein production in biotechnology and synthetic biology. Here, microfluidic single-cell analysis serves as the method of choice to investigate and unmask population heterogeneities in a dynamic and spatiotemporal fashion. In this study, we report on comparative microfluidic single-cell analyses of commonly used E. coli expression systems to uncover system-inherent specifications in the synthetic M9CA growth medium. To this end, the PT7lac/LacI, the PBAD/AraC and the Pm/XylS system were systematically analyzed in order to gain detailed insights into variations of growth behavior and expression phenotypes and thus to uncover individual strengths and deficiencies at the single-cell level. Specifically, we evaluated the impact of different system-specific inducers, inducer concentrations as well as genetic modifications that affect inducer-uptake and regulation of target gene expression on responsiveness and phenotypic heterogeneity. Interestingly, the most frequently applied expression system based on E. coli strain BL21(DE3) clearly fell behind with respect to expression homogeneity and robustness of growth. Moreover, both the choice of inducer and the presence of inducer uptake systems proved crucial for phenotypic heterogeneity. Conclusively, microfluidic evaluation of different inducible E. coli expression systems and setups identified the modified lacY-deficient PT7lac/LacI as well as the Pm/XylS system with conventional m-toluic acid induction as key players for precise and robust

  1. Physiological Response to a Simplified Veno-Venous Perfusion-Induced Systemic Hyperthermia System

    PubMed Central

    Ballard-Croft, Cherry; Wang, Dongfang; Jones, Cameron; Sumpter, L. Ryan; Zhou, Xiaoqin; Thomas, Joe; Topaz, Stephen; Zwischenberger, Joseph B.

    2014-01-01

    Our original venovenous perfusion-induced systemic hyperthermia (vv-PISH) system appeared to significantly improve lung cancer patient survival, but was too complex with numerous dialysis problems. We tested a simplified vv-PISH circuit that includes the AvalonElite double lumen cannula, a modified heat exchanger, water heater/cooler, and a centrifugal pump. The purpose of this study was to evaluate this simplified vv-PISH system (without hemodialyzer) and to investigate the physiological response to whole body hyperthermia in pigs. We tested our vv-PISH circuit in healthy female adult swine (n= 7, 55-68 kg). The therapeutic core temperature (42°C), calculated as mean of rectal, bladder, and esophageal temperatures, was achieved in six swine. A maximum difference of 0.5°C was observed between the individual temperature sensor readings, indicating homogeneous heat distribution. Heart rate and mean arterial pressure were transiently altered, but were safely managed. A significant elevation in pulmonary artery pressure occurred during the heating phase, resulting in one pig death. In all other pigs, pulmonary artery pressure returned to physiologic values during the therapeutic phase. Arterial blood electrolytes were maintained without the need of a dialyzer. Major organ function was within normal parameters. The simplified vv-PISH circuit reliably delivered the hyperthermic dose with no need of dialysis. PMID:23085942

  2. Novel microfluidic system for online monitoring of biofilm dynamics by electrical impedance spectroscopy and amperometry

    NASA Astrophysics Data System (ADS)

    Bruchmann, Julia; Sachsenheimer, Kai; Schwartz, Thomas; Rapp, Bastian E.

    2016-03-01

    Biofilm formation is ubiquitous in nature where microorganisms attach to surfaces and form highly adapted and protected communities. In technical and industrial systems like drinking water supply, food production or shipping industry biofilms are a major cause of product contamination, biofouling, and biocorrosion. Therefore, understanding of biofilm formation and means of preventing biofilm formation is important to develop novel biofilm treatment strategies. A system allowing directly online detection and monitoring biofilm formation is necessary. However, until today, there are little to none technical systems featuring a non-destructive real-time characterization of biofilm formation in a highthroughput manner. This paper presents such a microfluidic system based on electrical impedance spectroscopy (EIS) and amperomertic current measurement. The sensor consists of four modules, each housing 24 independent electrodes within 12 microfluidic channels. Attached biomass on the electrodes is monitored as increased inhibition in charge transfer by EIS and a change in metabolic activity is measured as change in produced electric current by amperometry. This modular sensor system is highly adaptable and suitable for a broad range of microbiological applications. Among others, biofilm formation processes can be characterized online, biofilm manipulation like inactivation or destabilization can be monitored in real-time and gene expression can be analyzed in parallel. The use of different electrode designs allows effective biofilm studies during all biofilm phases. The whole system was recently extended by an integrated pneumatic microfluidic pump which enables easy handling procedures. Further developments of this pumping module will allow a fully- automated computer-controlled valving and pumping.

  3. Automated and miniaturized detection of biological threats with a centrifugal microfluidic system

    NASA Astrophysics Data System (ADS)

    Mark, D.; van Oordt, T.; Strohmeier, O.; Roth, G.; Drexler, J.; Eberhard, M.; Niedrig, M.; Patel, P.; Zgaga-Griesz, A.; Bessler, W.; Weidmann, M.; Hufert, F.; Zengerle, R.; von Stetten, F.

    2012-06-01

    The world's growing mobility, mass tourism, and the threat of terrorism increase the risk of the fast spread of infectious microorganisms and toxins. Today's procedures for pathogen detection involve complex stationary devices, and are often too time consuming for a rapid and effective response. Therefore a robust and mobile diagnostic system is required. We present a microstructured LabDisk which performs complex biochemical analyses together with a mobile centrifugal microfluidic device which processes the LabDisk. This portable system will allow fully automated and rapid detection of biological threats at the point-of-need.

  4. Plant-in-chip: Microfluidic system for studying root growth and pathogenic interactions in Arabidopsis

    NASA Astrophysics Data System (ADS)

    Parashar, Archana; Pandey, Santosh

    2011-06-01

    We report a microfluidic platform for the hydroponic growth of Arabidopsis plants with high-resolution visualization of root development and root-pathogen interactions. The platform comprises a set of parallel microchannels with individual input/output ports where 1-day old germinated seedlings are initially placed. Under optimum conditions, a root system grows in each microchannel and its images are recorded over a 198-h period. Different concentrations of plant growth media show different root growth characteristics. Later, the developed roots are inoculated with two plant pathogens (nematodes and zoospores) and their physicochemical interactions with the live root systems are observed.

  5. Nucleic Acid-based Detection of Bacterial Pathogens Using Integrated Microfluidic Platform Systems

    PubMed Central

    Lui, Clarissa; Cady, Nathaniel C.; Batt, Carl A.

    2009-01-01

    The advent of nucleic acid-based pathogen detection methods offers increased sensitivity and specificity over traditional microbiological techniques, driving the development of portable, integrated biosensors. The miniaturization and automation of integrated detection systems presents a significant advantage for rapid, portable field-based testing. In this review, we highlight current developments and directions in nucleic acid-based micro total analysis systems for the detection of bacterial pathogens. Recent progress in the miniaturization of microfluidic processing steps for cell capture, DNA extraction and purification, polymerase chain reaction, and product detection are detailed. Discussions include strategies and challenges for implementation of an integrated portable platform. PMID:22412335

  6. A compact disk-like centrifugal microfluidic system for high-throughput nanoliter-scale protein crystallization screening.

    PubMed

    Li, Gang; Chen, Qiang; Li, Junjun; Hu, Xiaojian; Zhao, Jianlong

    2010-06-01

    A centrifuge-based microfluidic system has been developed that enables automated high-throughput and low-volume protein crystallizations. In this system, protein solution was automatically and accurately metered and dispensed into nanoliter-sized multiple reaction chambers, and it was mixed with various types of precipitants using a combination of capillary effect and centrifugal force. It has the advantages of simple fabrication, easy operation, and extremely low waste. To demonstrate the feasibility of this system, we constructed a chip containing 24 units and used it to perform lysozyme and cyan fluorescent protein (CyPet) crystallization trials. The results demonstrate that high-quality crystals can be grown and harvested from such a nanoliter-volume microfluidic system. Compared to other microfluidic technologies for protein crystallization, this microfluidic system allows zero waste, simple structure and convenient operation, which suggests that our microfluidic disk can be applied not only to protein crystallization, but also to the miniaturization of various biochemical reactions requiring precise nanoscale control. PMID:20459060

  7. Robust fluidic connections to freestanding microfluidic hydrogels

    PubMed Central

    Baer, Bradly B.; Larsen, Taylor S. H.

    2015-01-01

    Biomimetic scaffolds approaching physiological scale, whose size and large cellular load far exceed the limits of diffusion, require incorporation of a fluidic means to achieve adequate nutrient/metabolite exchange. This need has driven the extension of microfluidic technologies into the area of biomaterials. While construction of perfusable scaffolds is essentially a problem of microfluidic device fabrication, functional implementation of free-standing, thick-tissue constructs depends upon successful integration of external pumping mechanisms through optimized connective assemblies. However, a critical analysis to identify optimal materials/assembly components for hydrogel substrates has received little focus to date. This investigation addresses this issue directly by evaluating the efficacy of a range of adhesive and mechanical fluidic connection methods to gelatin hydrogel constructs based upon both mechanical property analysis and cell compatibility. Results identify a novel bioadhesive, comprised of two enzymatically modified gelatin compounds, for connecting tubing to hydrogel constructs that is both structurally robust and non-cytotoxic. Furthermore, outcomes from this study provide clear evidence that fluidic interconnect success varies with substrate composition (specifically hydrogel versus polydimethylsiloxane), highlighting not only the importance of selecting the appropriately tailored components for fluidic hydrogel systems but also that of encouraging ongoing, targeted exploration of this issue. The optimization of such interconnect systems will ultimately promote exciting scientific and therapeutic developments provided by microfluidic, cell-laden scaffolds. PMID:26045731

  8. Enhancement of antibody production by growth factor addition in perfusion and hollow-fiber culture systems.

    PubMed

    Omasa, T; Kobayashi, M; Nishikawa, T; Shioya, S; Suga, K; Uemura, S; Kitani, Y; Imamura, Y

    1995-12-20

    The effects of the high-molecular-weight growth factors, transferrin and bovine serum albumin (BSA), on antibody production were analyzed quantitatively in continuous hollow-fiber cultivation over a period of 60 days. Transferrin enhanced cell growth but had no significant effect on the specific antibody production rate, whereas BSA significantly enhanced antibody production. The antibody production rate was increased 4- and 14-fold respectively by feeding BSA at 2 and 5 g L(-1) into the EC side of the system (the side connected to the cell-containing outer part of the hollow-fiber unit) compared with the production achieved without BSA. Addition of 5 g L(1) BSA into the IC side of the system (the side connected to the inner part of the hollow-fiber unit) resulted in a 2.5-fold increase in the antibody production rate. The effect of BSA was also analyzed using the perfusion culture system with a separation unit. When fresh medium containing either 2 or 5 g L(-1) BSA was fed into the reactor, both the specific growth rate and specific death rate increased, while the specific antibody production rate was increased 2- and 25-fold, respectively, by feeding BSA at these two concentrations compared with no addition. Comparing the two systems, the increase in the antibody production rate achieved with the hollow-fiber system was threefold greater than that in the perfusion culture system with the same concentration of BSA feeding. (c) 1995 John Wiley & Sons, Inc. PMID:18623537

  9. Characterization of microfluidic human epidermal keratinocyte culture

    PubMed Central

    O’Neill, Adrian T.; Monteiro-Riviere, Nancy A.

    2008-01-01

    Human epidermal keratinocytes (HEK) are skin cells of primary importance in maintaining the body’s defensive barrier and are used in vitro to assess the irritation potential and toxicity of chemical compounds. Microfluidic systems hold promise for high throughput irritant and toxicity assays, but HEK growth kinetics have yet to be characterized within microscale culture chambers. This research demonstrates HEK patterning on microscale patches of Type I collagen within microfluidic channels and maintenance of these cells under constant medium perfusion for 72 h. HEK were shown to maintain 93.0%–99.6% viability at 72 h under medium perfusion ranging from 0.025–0.4 μl min−1. HEK maintained this viability while ∼100% confluent—a level not possible in 96 well plates. Microscale HEK cultures offer the ability to precisely examine the morphology, behavior and viability of individual cells which may open the door to new discoveries in toxicological screening methods and wound healing techniques. PMID:19002858

  10. High-Throughput Cytotoxicity Testing System of Acetaminophen Using a Microfluidic Device (MFD) in HepG2 Cells.

    PubMed

    Ju, Seon Min; Jang, Hyun-Jun; Kim, Kyu-Bong; Kim, Jeongyun

    2015-01-01

    A lab-on-a-chip (LOC) is a microfluidic device (MFD) that integrates several lab functions into a single chip of only millimeters in size. LOC provides several advantages, such as low fluidic volumes consumption, faster analysis, compactness, and massive parallelization. These properties enable a microfluidic-based high-throughput drug screening (HTDS) system to acquire cell-based abundant cytotoxicity results depending on linear gradient concentration of drug with only few hundreds of microliters of the drug. Therefore, a microfluidic device was developed containing an array of eight separate microchambers for cultivating HepG2 cells to be exposed to eight different concentrations of acetaminophen (APAP) through a diffusive-mixing-based concentration gradient generator. Every chamber array with eight different concentrations (0, 5.7, 11.4, 17.1, 22.8, 28.5, 34.2, or 40 mM) APAP had four replicating cell culture chambers. Consequently, 32 experimental results were acquired with a single microfluidic device experiment. The microfluidic high-throughput cytotoxicity device (μHTCD) and 96-well culture system showed comparable cytotoxicity results with increasing APAP concentration of 0 to 40 mM. The HTDS system yields progressive concentration-dependent cytotoxicity results using minimal reagent and time. Data suggest that the HTDS system may be applicable as alternative method for cytotoxicity screening for new drugs in diverse cell types. PMID:26241707

  11. A hybrid microfluidic system for regulation of neural differentiation in induced pluripotent stem cells.

    PubMed

    Hesari, Zahra; Soleimani, Massoud; Atyabi, Fatemeh; Sharifdini, Meysam; Nadri, Samad; Warkiani, Majid Ebrahimi; Zare, Mehrak; Dinarvand, Rassoul

    2016-06-01

    Controlling cellular orientation, proliferation, and differentiation is valuable in designing organ replacements and directing tissue regeneration. In the present study, we developed a hybrid microfluidic system to produce a dynamic microenvironment by placing aligned PDMS microgrooves on surface of biodegradable polymers as physical guidance cues for controlling the neural differentiation of human induced pluripotent stem cells (hiPSCs). The neuronal differentiation capacity of cultured hiPSCs in the microfluidic system and other control groups was investigated using quantitative real time PCR (qPCR) and immunocytochemistry. The functionally of differentiated hiPSCs inside hybrid system's scaffolds was also evaluated on the rat hemisected spinal cord in acute phase. Implanted cell's fate was examined using tissue freeze section and the functional recovery was evaluated according to the Basso, Beattie, and Bresnahan (BBB) locomotor rating scale. Our results confirmed the differentiation of hiPSCs to neuronal cells on the microfluidic device where the expression of neuronal-specific genes was significantly higher compared to those cultured on the other systems such as plain tissue culture dishes and scaffolds without fluidic channels. Although survival and integration of implanted hiPSCs did not lead to a significant functional recovery, we believe that combination of fluidic channels with nanofiber scaffolds provides a great microenvironment for neural tissue engineering, and can be used as a powerful tool for in situ monitoring of differentiation potential of various kinds of stem cells. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 1534-1543, 2016. PMID:26914600

  12. A PDMS-Based Cylindrical Hybrid Lens for Enhanced Fluorescence Detection in Microfluidic Systems

    PubMed Central

    Lin, Bor-Shyh; Yang, Yu-Ching; Ho, Chong-Yi; Yang, Han-Yu; Wang, Hsiang-Yu

    2014-01-01

    Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS) to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 μg/mL and 0.05 μg/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light. PMID:24531300

  13. Recent Results of the Investigation of a Microfluidic Sampling Chip and Sampling System for Hot Cell Aqueous Processing Streams

    SciTech Connect

    Julia Tripp; Jack Law; Tara Smith

    2013-10-01

    A Fuel Cycle Research and Development project has investigated an innovative sampling method that could evolve into the next generation sampling and analysis system for metallic elements present in aqueous processing streams. Initially sampling technologies were evaluated and microfluidics sampling chip technology was selected and tested. A conceptual design for a fully automated microcapillary-based system was completed and a robotic automated sampling system was fabricated. The mechanical and sampling operation of the completed sampling system was investigated. In addition, the production of a less expensive, mass produced sampling chip was investigated to avoid chip reuse thus increasing sampling reproducibility/accuracy. The microfluidic-based robotic sampling system’s mechanical elements were tested to ensure analytical reproducibility and the optimum robotic handling of microfluidic sampling chips.

  14. Portable laser speckle perfusion imaging system based on digital signal processor

    NASA Astrophysics Data System (ADS)

    Tang, Xuejun; Feng, Nengyun; Sun, Xiaoli; Li, Pengcheng; Luo, Qingming

    2010-12-01

    The ability to monitor blood flow in vivo is of major importance in clinical diagnosis and in basic researches of life science. As a noninvasive full-field technique without the need of scanning, laser speckle contrast imaging (LSCI) is widely used to study blood flow with high spatial and temporal resolution. Current LSCI systems are based on personal computers for image processing with large size, which potentially limit the widespread clinical utility. The need for portable laser speckle contrast imaging system that does not compromise processing efficiency is crucial in clinical diagnosis. However, the processing of laser speckle contrast images is time-consuming due to the heavy calculation for enormous high-resolution image data. To address this problem, a portable laser speckle perfusion imaging system based on digital signal processor (DSP) and the algorithm which is suitable for DSP is described. With highly integrated DSP and the algorithm, we have markedly reduced the size and weight of the system as well as its energy consumption while preserving the high processing speed. In vivo experiments demonstrate that our portable laser speckle perfusion imaging system can obtain blood flow images at 25 frames per second with the resolution of 640 × 480 pixels. The portable and lightweight features make it capable of being adapted to a wide variety of application areas such as research laboratory, operating room, ambulance, and even disaster site.

  15. Portable laser speckle perfusion imaging system based on digital signal processor.

    PubMed

    Tang, Xuejun; Feng, Nengyun; Sun, Xiaoli; Li, Pengcheng; Luo, Qingming

    2010-12-01

    The ability to monitor blood flow in vivo is of major importance in clinical diagnosis and in basic researches of life science. As a noninvasive full-field technique without the need of scanning, laser speckle contrast imaging (LSCI) is widely used to study blood flow with high spatial and temporal resolution. Current LSCI systems are based on personal computers for image processing with large size, which potentially limit the widespread clinical utility. The need for portable laser speckle contrast imaging system that does not compromise processing efficiency is crucial in clinical diagnosis. However, the processing of laser speckle contrast images is time-consuming due to the heavy calculation for enormous high-resolution image data. To address this problem, a portable laser speckle perfusion imaging system based on digital signal processor (DSP) and the algorithm which is suitable for DSP is described. With highly integrated DSP and the algorithm, we have markedly reduced the size and weight of the system as well as its energy consumption while preserving the high processing speed. In vivo experiments demonstrate that our portable laser speckle perfusion imaging system can obtain blood flow images at 25 frames per second with the resolution of 640 × 480 pixels. The portable and lightweight features make it capable of being adapted to a wide variety of application areas such as research laboratory, operating room, ambulance, and even disaster site. PMID:21198054

  16. Detection and analysis of temperature-sensitive dermal blood perfusion dynamics and distribution by a hybrid camera system.

    PubMed

    Blanik, N; Paul, M; Blazek, V; Leonhardt, S

    2015-08-01

    In this paper we present an application of two optical imaging modalities for non-invasive assessment of dermal perfusion. This hybrid setup consists of a photo-plethysmographic camera sensing in the visible spectrum and a thermal camera sensing in the infrared-C-band. This allows to combine the information of both sources complementarily: The extracted perfusion index as well as the skin surface temperature. The feasibility of the presented system is tested in two studies with local temperature stress on the forehead of a subject. In the first, a local cooling on the subject's forehead is monitored and further analyzed. In the second, skin perfusion reactions to heat are considered. For both experiments the results are compared to baseline measurements and non-affected areas in the field of view of the cameras. As results, the dependencies between temperature and perfusion change are presented. Further, influences of the stressor can be visualized in functional mappings of calculated perfusion indices. For the performed test, a linear correlation between temperature and perfusion change is obtained. PMID:26736773

  17. Methods, microfluidic devices, and systems for detection of an active enzymatic agent

    SciTech Connect

    Sommer, Gregory J; Hatch, Anson V; Singh, Anup K; Wang, Ying-Chih

    2014-10-28

    Embodiments of the present invention provide methods, microfluidic devices, and systems for the detection of an active target agent in a fluid sample. A substrate molecule is used that contains a sequence which may cleave in the presence of an active target agent. A SNAP25 sequence is described, for example, that may be cleaved in the presence of Botulinum Neurotoxin. The substrate molecule includes a reporter moiety. The substrate molecule is exposed to the sample, and resulting reaction products separated using electrophoretic separation. The elution time of the reporter moiety may be utilized to identify the presence or absence of the active target agent.

  18. Sensing Characteristics of Shear-Mode AlN Solidly Mounted Resonators with a Silicone Microfluidic System in Viscous Media

    NASA Astrophysics Data System (ADS)

    Xiong, Juan; Guo, Peng; Sun, Xi-Liang; Wang, Sheng-Fu; Hu, Ming-Zhe; Gu, Hao-Shuang

    2014-02-01

    AlN solidly mounted resonators with silicone microfluidic systems vibrating in shear mode are fabricated and characterized. The fabrication process is compatible with integrated circuits and the c-axis tilted AlN films are deposited, which allow in-liquid operation through excitation of the shear mode. The silicone microfluidic system is mounted on top of the sensor chip to transport the analyses and confine the flow to the active area. The properties of sensor operation in air, deionized water, ethanol, isopropanol, 80% glycol aqueous solution, glycol, and olive oil are characterized. The effects of different viscosities on the resonance frequency shift and Q-factor of the sensor have been discussed. The sensitivity and Q value in glycol of the sensor are 1.52 MHz cm2/μg and around 60, respectively. The results indicate the potential of a highly sensitive microfluidic sensor system for the applications in viscous media.

  19. DropBot: An open-source digital microfluidic control system with precise control of electrostatic driving force and instantaneous drop velocity measurement

    SciTech Connect

    Fobel, Ryan; Donnelly Centre for Cellular and Biomolecular Research, 160 College St., Toronto, Ontario M5S 3E1 ; Fobel, Christian; Wheeler, Aaron R.; Donnelly Centre for Cellular and Biomolecular Research, 160 College St., Toronto, Ontario M5S 3E1; Department of Chemistry, University of Toronto, 80 St. George St., Toronto, Ontario M5S 3H6

    2013-05-13

    We introduce DropBot: an open-source instrument for digital microfluidics (http://microfluidics.utoronto.ca/dropbot). DropBot features two key functionalities for digital microfluidics: (1) real-time monitoring of instantaneous drop velocity (which we propose is a proxy for resistive forces), and (2) application of constant electrostatic driving forces through compensation for amplifier-loading and device capacitance. We anticipate that this system will enhance insight into failure modes and lead to new strategies for improved device reliability, and will be useful for the growing number of users who are adopting digital microfluidics for automated, miniaturized laboratory operation.

  20. DropBot: An open-source digital microfluidic control system with precise control of electrostatic driving force and instantaneous drop velocity measurement

    NASA Astrophysics Data System (ADS)

    Fobel, Ryan; Fobel, Christian; Wheeler, Aaron R.

    2013-05-01

    We introduce DropBot: an open-source instrument for digital microfluidics (http://microfluidics.utoronto.ca/dropbot). DropBot features two key functionalities for digital microfluidics: (1) real-time monitoring of instantaneous drop velocity (which we propose is a proxy for resistive forces), and (2) application of constant electrostatic driving forces through compensation for amplifier-loading and device capacitance. We anticipate that this system will enhance insight into failure modes and lead to new strategies for improved device reliability, and will be useful for the growing number of users who are adopting digital microfluidics for automated, miniaturized laboratory operation.

  1. Automatic microfluidic fluorescence-array measurement system for detecting organic phosphate.

    PubMed

    Chang, Hsing-Cheng; Lin, Jung-Chin; Lin, Shyan-Lung; Chang, I-Nan; Lin, Chern-Sheng; Chen, Shi-Yao

    2015-01-01

    In this study, an automatic microfluidic fluorescence-array measurement system is developed to detect the concentration of organic phosphate based on the luminol-hydrogen peroxide catalytic fluorescent mechanism. Not only sample quantity and cost can be reduced, but also detection time, accuracy and precision can be improved in the system. The system is composed of a CCD image module, a stepper motor with driver, a microfluidic fluorescence array, a background light elimination module, and a dynamic image-analyzed interface. The pesticides of chlorpyrifos and fenitrothion of organic phosphate are chosen as experimental samples. Only a 2.5 μ l quantity of sample is required to have a fast response time of 1.4 second. Experimental results show that the sensitivities of chlorpyrifos and fenitrothion are 1.88 V/ppm in the range of 0.166 ∼ 10 ppm with averaged error of 1.66% and 0.32 V/ppm in the range of 0.03 ∼ 10 ppm with averaged error of 1.68% respectively. The organophosphorus effective detection range of the developed system covers the legal prescription for pesticide residues. PMID:26409537

  2. Quantitative myocardial perfusion imaging in a porcine ischemia model using a prototype spectral detector CT system.

    PubMed

    Fahmi, Rachid; Eck, Brendan L; Levi, Jacob; Fares, Anas; Dhanantwari, Amar; Vembar, Mani; Bezerra, Hiram G; Wilson, David L

    2016-03-21

    We optimized and evaluated dynamic myocardial CT perfusion (CTP) imaging on a prototype spectral detector CT (SDCT) scanner. Simultaneous acquisition of energy sensitive projections on the SDCT system enabled projection-based material decomposition, which typically performs better than image-based decomposition required by some other system designs. In addition to virtual monoenergetic, or keV images, the SDCT provided conventional (kVp) images, allowing us to compare and contrast results. Physical phantom measurements demonstrated linearity of keV images, a requirement for quantitative perfusion. Comparisons of kVp to keV images demonstrated very significant reductions in tell-tale beam hardening (BH) artifacts in both phantom and pig images. In phantom images, consideration of iodine contrast to noise ratio and small residual BH artifacts suggested optimum processing at 70 keV. The processing pipeline for dynamic CTP measurements included 4D image registration, spatio-temporal noise filtering, and model-independent singular value decomposition deconvolution, automatically regularized using the L-curve criterion. In normal pig CTP, 70 keV perfusion estimates were homogeneous throughout the myocardium. At 120 kVp, flow was reduced by more than 20% on the BH-hypo-enhanced myocardium, a range that might falsely indicate actionable ischemia, considering the 0.8 threshold for actionable FFR. With partial occlusion of the left anterior descending (LAD) artery (FFR  <  0.8), perfusion defects at 70 keV were correctly identified in the LAD territory. At 120 kVp, BH affected the size and flow in the ischemic area; e.g. with FFR [Formula: see text] 0.65, the anterior-to-lateral flow ratio was 0.29  ±  0.01, over-estimating stenosis severity as compared to 0.42  ±  0.01 (p  <  0.05) at 70 keV. On the non-ischemic inferior wall (not a LAD territory), the flow ratio was 0.50  ±  0.04 falsely indicating an actionable ischemic condition

  3. Quantitative myocardial perfusion imaging in a porcine ischemia model using a prototype spectral detector CT system

    NASA Astrophysics Data System (ADS)

    Fahmi, Rachid; Eck, Brendan L.; Levi, Jacob; Fares, Anas; Dhanantwari, Amar; Vembar, Mani; Bezerra, Hiram G.; Wilson, David L.

    2016-03-01

    We optimized and evaluated dynamic myocardial CT perfusion (CTP) imaging on a prototype spectral detector CT (SDCT) scanner. Simultaneous acquisition of energy sensitive projections on the SDCT system enabled projection-based material decomposition, which typically performs better than image-based decomposition required by some other system designs. In addition to virtual monoenergetic, or keV images, the SDCT provided conventional (kVp) images, allowing us to compare and contrast results. Physical phantom measurements demonstrated linearity of keV images, a requirement for quantitative perfusion. Comparisons of kVp to keV images demonstrated very significant reductions in tell-tale beam hardening (BH) artifacts in both phantom and pig images. In phantom images, consideration of iodine contrast to noise ratio and small residual BH artifacts suggested optimum processing at 70 keV. The processing pipeline for dynamic CTP measurements included 4D image registration, spatio-temporal noise filtering, and model-independent singular value decomposition deconvolution, automatically regularized using the L-curve criterion. In normal pig CTP, 70 keV perfusion estimates were homogeneous throughout the myocardium. At 120 kVp, flow was reduced by more than 20% on the BH-hypo-enhanced myocardium, a range that might falsely indicate actionable ischemia, considering the 0.8 threshold for actionable FFR. With partial occlusion of the left anterior descending (LAD) artery (FFR  <  0.8), perfusion defects at 70 keV were correctly identified in the LAD territory. At 120 kVp, BH affected the size and flow in the ischemic area; e.g. with FFR ≈ 0.65, the anterior-to-lateral flow ratio was 0.29  ±  0.01, over-estimating stenosis severity as compared to 0.42  ±  0.01 (p  <  0.05) at 70 keV. On the non-ischemic inferior wall (not a LAD territory), the flow ratio was 0.50  ±  0.04 falsely indicating an actionable ischemic condition in a healthy

  4. A Scalable Perfusion Culture System with Miniature Peristaltic Pumps for Live-Cell Imaging Assays with Provision for Microfabricated Scaffolds

    PubMed Central

    Balakrishnan, Sreenath; Suma, M.S.; Raju, Shilpa R.; Bhargav, Santosh D.B.; Arunima, S.; Das, Saumitra

    2015-01-01

    Abstract We present a perfusion culture system with miniature bioreactors and peristaltic pumps. The bioreactors are designed for perfusion, live-cell imaging studies, easy incorporation of microfabricated scaffolds, and convenience of operation in standard cell culture techniques. By combining with miniature peristaltic pumps—one for each bioreactor to avoid cross-contamination and to maintain desired flow rate in each—we have made a culture system that facilitates perfusion culture inside standard incubators. This scalable system can support multiple parallel perfusion experiments. The major components are fabricated by three-dimensional printing using VeroWhite, which we show to be amenable to ex vivo cell culture. Furthermore, the components of the system can be reused, thus making it economical. We validate the system and illustrate its versatility by culturing primary rat hepatocytes, live imaging the growth of mouse fibroblasts (NIH 3T3) on microfabricated ring-scaffolds inserted into the bioreactor, performing perfusion culture of breast cancer cells (MCF7), and high-magnification imaging of hepatocarcinoma cells (HuH7). PMID:26309810

  5. SU-E-QI-06: Design and Initial Validation of a Precise Capillary Phantom to Test Perfusion Systems

    SciTech Connect

    Wood, R; Iacobucci, G; Khobragade, P; Ying, L; Snyder, K; Wack, D; Rudin, S; Ionita, C

    2014-06-15

    Purpose: To design a precise perfusion phantom mimicking capillaries of the brain vasculature which could be used to test various perfusion protocols and algorithms which generate perfusion maps. Methods: A perfusion phantom was designed in Solidworks and built using additive manufacturing. The phantom was an overall cylindrical shape of diameter and height 20mm and containing capillaries of 200μm or 300μm which were parallel and in contact making up the inside volume where flow was allowed. We created a flow loop using a peristaltic pump and contrast agent was injected manually. Digital Subtraction Angiographic images and low contrast images with cone beam CT were acquired after the contrast was injected. These images were analyzed by our own code in LabVIEW software and Time-Density Curve, MTT and TTP was calculated. Results: Perfused area was visible in the cone beam CT images; however, individual capillaries were not distinguishable. The Time-Density Curve acquired was accurate, sensitive and repeatable. The parameters MTT, and TTP offered by the phantom were very sensitive to slight changes in the TDC shape. Conclusion: We have created a robust calibrating model for evaluation of existing perfusion data analysis systems. This approach is extremely sensitive to changes in the flow due to the high temporal resolution and could be used as a golden standard to assist developers in calibrating and testing of imaging perfusion systems and software algorithms. Supported by NIH Grant: 2R01EB002873 and an equipment grant from Toshiba Medical Systems Corporation.

  6. Changes in mixed-function oxidase system in the perfused liver of the cold-acclimated rat

    NASA Astrophysics Data System (ADS)

    Takano, T.; Miyazaki, Y.; Motohashi, Y.; Yamada, K.

    1986-09-01

    Changes in the hepatic cytochrome P-450-dependent drug-metabolizing system were studied in perfused livers obtained from cold-acclimated male Wistar rats after 30 days of cold exposure (4‡C) when using hexobarbital as a substrate. In fasted animals the cold-acclimated rats showed higher levels of hexobarbital metabolic rates compared to control rats, but there was no significant difference in fed animals. The maximum rates of hexobarbital metabolism produced by xylitol perfusion were also significantly higher in the perfused liver of cold-acclimated rats. It was concluded that the function of the cytochrome P-450 system for hexobarbital in cold-acclimated rats changed due to both an increase in the activity of the cytochrome P-450 system and to changes in regulation of the cytochrome P-450 system by the supply of reducing equivalents.

  7. A microfluidic manifold with a single pump system to generate highly mono-disperse alginate beads for cell encapsulation

    PubMed Central

    Kim, Choong; Park, Juyoung

    2014-01-01

    Cell encapsulation technology is a promising strategy applicable to tissue engineering and cell therapy. Many advanced microencapsulation chips that function via multiple syringe pumps have been developed to generate mono-disperse hydrogel beads encapsulating cells. However, their operation is difficult and only trained microfluidic engineers can use them with dexterity. Hence, we propose a microfluidic manifold system, driven by a single syringe pump, which can enable the setup of automated flow sequences and generate highly mono-disperse alginate beads by minimizing disturbances to the pump pressure. The encapsulation of P19 mouse embryonic carcinoma cells and embryonic body formation are demonstrated to prove the efficiency of the proposed system. PMID:25587376

  8. A microfluidic chemical/biological sensing system based on membrane dissolution and optical absorption

    NASA Astrophysics Data System (ADS)

    Sridharamurthy, Sudheer S.; Dong, Liang; Jiang, Hongrui

    2007-01-01

    A microfluidic system to sense chemical and biological analytes using membranes dissolvable by the analyte is demonstrated. The scheme to detect the dissolution of the membrane is based on the difference in optical absorption of the membrane and the fluidic sample being assayed. The presence of the analyte in the sample chemically cleaves the membrane and causes the sample to flow into the membrane area. This causes a change in the optical absorption of the path between the light source and detector. A device comprising the microfluidic channels and the membrane is microfabricated using liquid-phase photopolymerization. A light emitting diode (LED) and a detector with an integrated amplifier are positioned and aligned on either side of the device. The state of the membrane is continuously monitored after introducing the sample. The temporal dissolution characteristics of the membrane are extracted in terms of the output voltage of the detector as a function of time. This is used to determine the concentration of the analyte. The absorption spectra of the membrane and fluidic sample are studied to determine the optimal wavelength that provides the maximum difference in absorbance between the membrane and the sample. In this work, the dissolution of a poly(acrylamide) hydrogel membrane in the presence of a reducing agent (dithiothreitol—DTT) is used as a model system. For this system, with 1 M DTT, complete membrane dissolution occurred after 65 min.

  9. Perfusion Assessment with the SPY System after Arterial Venous Reversal for Upper Extremity Ischemia

    PubMed Central

    2014-01-01

    Background: The timing and pattern of reperfusion following arterial- venous reversal (AVR) in patients with terminal ischemia of an upper extremity is not well understood. Methods: The current case series describes the timing and pattern of reperfusion observed in patients with terminal upper extremity ischemia who underwent AVR and repeated postoperative indocyanine green (ICG) angiography between 2004 and 2009. For all included patients, the SPY Near-Infrared Perfusion Assessment System permitted visualization of ICG-labeled blood flow for 60-second sampling periods at scheduled postoperative time points; outflow and rate and amplitude of inflow were objectively quantified with SPY-Q Analysis Toolkit image analysis software. Results: The series comprised 6 male patients (mean age, 46 years) who presented with upper extremity ischemia related to hypothenar hammer syndrome (n = 2), embolism with patent foramen ovale (n = 2), atherosclerosis (n = 1), and avulsion amputation of the thumb (n = 1); the patient with the avulsion amputation was diagnosed with thromboangiitis obliterans at the time of replantation. AVR was successful in all 6 patients. In 5 of 6 patients, ICG angiography and SPY-based visualization/quantification showed that venous outflow and arterial inflow gradually normalized (versus unaffected digits) between postoperative days (PODs) 0 and 3 and was maintained at long-term follow-up (≥3 months); for the patient who underwent thumb replantation, perfusion normalized between POD 3 and month 5 follow-up. Conclusions: AVR effectively reestablished blood flow in patients with terminal upper extremity ischemia. ICG angiography with SPY technology revealed that, in most cases, kinetic curves, timing, and patterns of perfusion gradually normalized over several PODs. PMID:25426368

  10. Magnetic manipulation of superparamagnetic nanoparticles in a microfluidic system for drug delivery applications

    NASA Astrophysics Data System (ADS)

    Agiotis, L.; Theodorakos, I.; Samothrakitis, S.; Papazoglou, S.; Zergioti, I.; Raptis, Y. S.

    2016-03-01

    Magnetic nanoparticles (MNPs), such as superparamagnetic iron oxide nanoparticles (SPIONS), have attracted major interest, due to their small size and unique magnetic properties, for drug delivery applications. In this context, iron oxide nanoparticles of magnetite (Fe3O4) (150 nm magnetic core diameter), were used as drug carriers, aiming to form a magnetically controlled nano-platform. The navigation capabilities of the iron oxide nanoparticles in a microfluidic channel were investigated by simulating the magnetic field and the magnetic force applied on the magnetic nanoparticles inside a microfluidic chip. The simulations have been performed using finite element method (ANSY'S software). The optimum setup which intends to simulate the magnetic navigation of the nanoparticles, by the use of MRI-type fields, in the human circulatory system, consists of two parallel permanent magnets to produce a homogeneous magnetic field, in order to ensure the maximum magnetization of the magnetic nanoparticles, an electromagnet for the induction of the magnetic gradients and the creation of the magnetic force and a microfluidic setup so as to simulate the blood flow inside the human blood vessels. The magnetization of the superparamagnetic nanoparticles and the consequent magnetic torque developed by the two permanent magnets, together with the mutual interactions between the magnetized nanoparticles lead to the creation of rhabdoid aggregates in the direction of the homogeneous field. Additionally, the magnetic gradients introduced by the operation of the electromagnet are capable of directing the aggregates, as a whole, to the desired direction. By removing the magnetic fields, the aggregates are disrupted, due to the super paramagnetic nature of the nanoparticles, avoiding thus the formation of undesired thrombosis.