Rozprawy doktorskie na temat „Microfluidic devices”

Microfluidics have been created to acquire, operate, and process complex fluids in extremely tiny volumes with high efficiency and high speed, and without the requirement for an experienced operator. In addition, microfluidic systems also enable miniaturization and incorporation of different complex functions, which can help bring intricate diagnostic tools out of the laboratories. Ideally, these systems should be inexpensive, precise, reliable, robust, and well-suited to the medical diagnostic systems. Most of the microfluidic devices reported previously were based on devices made of polydimethylsiloxane (PDMS). PDMS is a material that dissolves in many common organic solvents. Meanwhile, it is also prone to absorb small molecules like the proteins, which is detrimental to a stable and reliable result. Current work focuses on bioassays that are badly needed in our life and these bioassays are addressed based on microfluidic platform with different materials. The translation of microfluidic technology into large scale implementations highly relies on new materials that address the limitations of PDMS. Firstly, we fabricated two different microfluidic platforms for rapid antimicrobial susceptibility testing (AST). One was made of hydrogel, and the bacterial cells were cultured on the top of the device; the other was of polypropylene (PP), and bacterial cells were cultured inside the microchannels. Meanwhile, we developed a novel "barcode" sensor, a microscope-free method for cell accumulation and cell counting, as the downstream of the PP-based chips. As a result, AST can be accomplished simply through an application on a mobile phone rather than using an expensive and sophisticated microscope. Secondly, we presented a self-contained paper-based system for lead(II) ion detection based on G-quadruplex-based luminescence switch-on assay, comprising a novel type of paper-based chip and a matching portable device. Different from the reported paper-based devices, the paper substrate we chose was art paper, which is used for printing magazines. This type of paper could prevent the absorption of liquid into the paper matrix and hold the liquid in place for a period of time; and it could also be used for temporary liquid containing like a plastic substrate (such as polypropylene (PP) and polystyrene (PS)), but the surface of the paper is inherently hydrophilic. In such a design, liquid drops are suspended on the surface of the device in designed reservoirs, rather than absorbed into the paper; when the chip is tilted, the liquid drops will move to other reservoirs according to the guidance of channels defined on the surface. To differentiate it from reported μPAD devices that are fabricated with water-permeable paper, we name this new type of paper-based devices suspending-droplet mode paper-based microfluidic devices (SD-μPAD). Different from the conventional μPADs that use capillary force to drive liquid, our SD-μPADs uses wetting and gravity as driving force. To fabricate the superhydrophobic pattern on the paper device, we developed a new microcontact printing-based method to produce inexpensive and precisely patterned superhydrophobic coating on paper. The coating material is poly(dimethylsiloxane) (PDMS), a hydrophobic and transparent silicone that has long been used for fabricating microfluidic devices. Importantly, the negative-relief stamp we used is made of Teflon, a non-stick polymer, so that the PDMS-coated paper could be peeled from the stamp flawlessly. After such fabrication process, the stamped area of the paper is coated with a textured PDMS layer that is decorated with arrays of micropillars, which could provide superhydrophobic effect and most effectively hold the droplets in place; the remaining area of the paper is still hydrophilic. As a demonstration of this new design, we developed a method using the reaction characteristics of iridium(III) complex for rapid, onsite detection of lead(II) ions in liquid samples. As the reagents have already been loaded onto the paper device during fabrication, the only reagent the users need to add is water. Because of the large Stokes shift of the iridium(III) complex probe, inexpensive optical filters can be employed, and we were able to make an inexpensive, battery-powered compact device for routine portable detection using a smartphone as a detector, allowing the rapid analysis and interpretation of results on site as well as the automatic dissemination of data to professional institutes, including tests even in poor rural areas in developing countries. Thirdly, we upgraded our suspending-droplet mode paper-based microfluidic device (SD-μPAD), which is used for the detection of lead(II) ions in liquid solution. The reason is that our paper-based SD chips are not suitable for long reaction process (> 20 min) detection of biomolecules due to the potential permeation and contaminating problems of art papers. Hence, we chose polypropylene (PP), a hydrophobic, cheap, and thermal stable material (< 110°C), as the material for the fabrication of the SD microfluidic chip. We established a convenient, low-cost, portable and reliable platform for monitoring VEGF165 accurately, which can be applied for point-of-care (POC) testing. In this project, we also employed the label-free oligonucleotide-based luminescence switch-on assay on the microfluidic platform, which possesses the advantages of high sensitivity and high selectivity. Based on the detection of VEGF165 in a three-step reaction process, we adopted a new design for the droplet transfer throughout the channels. This design could migrate the droplet through the chambers via controlling the orientation of the chip, which systematically combined the superhydrophobic force of the coating, the gravity of the droplet and the surface tension between PP and droplet. Therefore, traditional micro pump could be avoided and the total cost for the device could be substantially reduced. In addition, we developed an automatic, matched and portable device for the detection of VEGF165, which assembled by a rotatable chip holder, a UV lamp, a filter, and a camera. Finally, we developed a new whole Teflon membrane-based chip for the aptamer screening. Our article "Whole-Teflon microfluidic chips" introduced the fabrication of a microfluidic device entirely using Teflon materials, one group of the most inert materials in the world. It was a successful and representative introduction of new materials into the fabrication of microfluidic devices, which show dramatically greater anti-fouling performance. However, even such device was inadequate for current purpose, as it is rigid and lacks convenient valve control functions for particle suspensions used in systematic evolution of ligands by exponential enrichment (SELEX). For this project, we propose a SMART screening strategy based on a highly integrated microfluidic chip. This new type of whole-Teflon devices, which are made of flexible Teflon membranes, offering convenient valving control for the whole SELEX process to be performed on chip and fulfilling the anti-fouling requirement in the meantime. The SELEX cycles including positive and negative selections could be automatically performed inside tiny-size microchambers on a microchip, and the enrichment is real-time monitored. The selection cycles would be ended after the resulted signal of the aptamers with high specificity reached a plateau, or no target aptamer is captured after a number of cycles of enrichment. Owning to the antifouling property of the chip materials, the loss of the sample is tremendously reduced. The SMART platform therefore is not only free of complicated manual operations, but also high-yield and well reproducible over conventional methods

Microfluidics is the science and technology of manipulating and analysing small amounts of liquid. Microfluidics has several advantages including small sample volume, small footprint, being cheap, portable, and precise. Microfluidics has applications in a wide range of areas such as in chemistry, electronics, and most importantly in biological sciences. Microfluidic functions are greatly influenced by the geometry and dimensions of the microchannels. The main challenge facing microfluidics is that once the conventional rigid microfluidic device is fabricated, its dimensions cannot be changed or modified. To overcome this problem, we proposed the concept of flexible and stretchable microfluidics. Stretchable microfluidics allows flexible devices to change their dimensions and thus enabling new functionalities. This thesis aims to (i) understand the fundamentals of flexible microfluidics, (ii) design and fabricate a new generation of stretchable microfluidic devices with tuneable dimensions, and (iii) apply stretchable microfluidics to three main handling tasks of separation mixing and trapping. Our main goal is to evaluate how the dimensions of different types of microfluidic devices alter under elongation and how these dimensional changes influence its functions. In this thesis, following a comprehensive introduction in chapter 1, a thorough literature review over flexible microfluidics is provided in chapter 2. The review covers three main areas of flexible microfluidics including materials, effect of flexibility on microfluidic functions, and the current applications and future perspectives of flexible microfluidics. Chapter 3 and 4 investigate the effect of stretchability on inertial microfluidics. Inertial microfluidics is a promising approach for particle separation. The current obstacle of inertial microfluidics in biological applications is the broad size distribution of biological microparticles. Rigid microfluidic devices work well for a narrow range of particle sizes. For focusing and separating a new set of particles, troublesome and time-consuming design, fabrication, testing, and optimization procedures are needed. Thus, a stretchable a microfluidic device with tuneable dimensions was fabricated and studied in chapter 3. By changing the channel dimensions under elongation, the device could be adapted to different particle sizes and flow rate ratios. Stretching the device significantly improved the focusing and separation efficiency of the specific particle sizes. In chapter 4, we focused on the application of stretchable inertial microfluidics for cancer detection. The performance of the stretchable device was verified by isolating cancer cells from WBCs and from whole blood with high recovery rates and purities. Chapter 5 studies the effect of stretchability on micromixing. A micromixer is an indispensable component in miniaturised platforms for chemical, biochemical, and biomedical applications. Mixing in microscale is challenging due to the laminar flow associated with low Reynolds numbers. This chapter reports a stretchable micromixer with dynamically tuneable channel dimensions. Periodic elongation of the stretchable micromixer results in mixing disturbance in intermediate Reynolds numbers. Periodically stretching the device changes the channel geometry and dimensions leading to dynamically evolving secondary and main flows. We evaluated the performance of this stretchable micromixer both experimentally and numerically. Chapter 6 reports a stretchable microtrapper. Microfluidic technologies have been widely used for single-cell trapping. However, there are no robust methods for the facile release of the captured cells for subsequent studies. Therefore, we developed a stretchable microfluidic cell trapper for easy on-demand release of cells in a deterministic manner. By tunning the horizontal elongation of the device, the gap at the bottom of the traps widened and provided ample space for releasing particle/cell with sizes of interest. The proposed stretchable micro trapper demonstrated a deterministic recovery of the captured cells by adjusting the elongation length of the device. Flexible and stretchable microfluidic devices with tuneable dimensions were introduced and studied extensively in this thesis. We showed that by applying stretchability to microfluidic functions including inertial microfluidics, micromixing, and single cell studies, several drawbacks associated with fixed dimensions were addressed and recovered. We believe that flexible and stretchable microfluidics is a new research direction of microfluidics.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Environment and Sc
Science, Environment, Engineering and Technology
Full Text

My research broadly covers various important aspects of microfluidic devices and biosensors. Specifically, this dissertation reports: (1) a new and effective room temperature method of bonding polydimethylsiloxane (PDMS) microfluidics to substrates such as silicon and glass, (2) a new microfluidic pump concept and implementation specifically designed to repeatedly drive a small sample volume (<1 µL) very rapidly (~500 µL/min) through a sensor-containing flow channel to significantly decrease sensor response time through advection-driven rather than diffusion-driven mass transport, (3) use of a new microfluidic material based on polyethylene glycol diacrylate (PEGDA) to implement impedance-based dynamic nanochannel sensors for protein sensing, and (4) an investigation of galvanoluminescence and how to avoid it for conditions important to fluorescence-based dielectrophoresis (DEP) microfluidic biosensors. Over the last decade, the Nordin research group has developed a lab-on-a-chip (LOC) biosensor based on silicon photonic microcantilever arrays integrated with polydimethylsiloxane (PDMS) microfluidics for protein biomarker detection. Integration requires reliable bonding at room temperature with adequate bond strength between the PDMS element and microcantilever sensor substrate. The requirement for a room temperature process is particularly critical because microcantilevers must be individually functionalized with antibody-based receptor molecules prior to bonding and cannot withstand significant heating after functionalization. I developed a new room temperature bonding method using PDMS curing agent as an intermediate adhesive layer. Two curing agents (Sylgard 184 and 182) were compared, as well as an alternate UV curable adhesive (NOA 75). The bond strength of Sylgard 184 was found to be stronger than Sylgard 182 under the same curing conditions. Overnight room temperature curing with Sylgard 184 yields an average burst pressure of 433 kPa, which is more than adequate for many PDMS sensor devices. In contrast, UV curable epoxy required a 12 hour bake at 50 °C to achieve maximum bond strength, which resulted in a burst pressure of only 124 kPa. In many biosensing scenarios it is desirable to use a small sample volume (<1 µL) to detect small analyte concentrations in as short a time as possible. I report a new microfluidic pump to address this need, which we call a reflow pump. It is designed to rapidly pump a small sample volume back and forth in a flow channel. Ultimately, the flow channel would contain functionalized sensor surfaces. The rapid flow permits use of advection-driven mass transport to the sensor surfaces to dramatically reduce sensor response times compared to diffusion-based mass transport. Normally such rapid flow would have the effect of decreasing the fraction of analyte molecules in the volume that would see the sensor surfaces. By configuring the pump to reflow fluid back and forth in the flow channel, the analyte molecules in the small sample volume are used efficiently in that they have many opportunities to make it to the sensor surfaces. I describe a 3-layer PDMS reflow pump that pumps 300 nL of fluid at 500 µL/min for 15 psi actuation pressure, and demonstrate a new two-layer configuration that significantly simplifies pump fabrication. Impedance-based nanochannel sensors operate on the basis of capturing target molecules in nanochannels such that impedance through the nanochannels is increased. While simple in concept, the response time can be quite long (8~12 hours) because the achievable flow rate through a nanochannel is very limited. An approach to dramatically increase the flow rate is to form nanochannels only during impedance measurements, and otherwise have an array of nanotrenches on the surface of a conventional microfluidic flow channel where they are exposed to normal microfluidic flow rates. I have implemented such a dynamic nanochannel approach with a recently-developed microfluidic material based polyethylene glycol diacrylate (PEGDA). I present the design, fabrication, and testing of PEGDA dynamic nanochannel array sensors, and demonstrate an 11.2 % increase in nanochannel impedance when exposed to 7.2 µM bovine serum albumin (BSA) in phosphate buffered saline (PBS). Recently, LOC biosensors for cancer cell detection have been demonstrated based on a combination of dielectrophoresis (DEP) and fluorescence detection. For fluorescence detection it is critical to minimize other sources of light in the system. However, reported devices use a non-noble metal electrode, indium tin oxide (ITO), to take advantage of its optical transparency. Unfortunately, use of non-noble metal electrodes can result in galvanoluminescence (GL) in which the AC voltage applied to the electrodes to achieve DEP causes light emission, which can potentially confound the fluorescence measurement. I designed and fabricated two types of devices to examine and identify conditions that lead to GL. Based on my observations, I have developed a method to avoid GL that involves measuring the impedance spectrum of a DEP device and choosing an operating frequency in the resistive portion of the spectrum. I also measure the emission spectrum of twelve salt solutions, all of which exhibited broadband GL. Finally, I show that in addition to Au, Cr and Ni do not exhibit GL, are therefore potentially attractive as low cost DEP electrode materials.

This work is a study to understand the various aspects of a microfluidic device. In the first half we take the role of an end user, experimenting to learn how best to use the device efficiently. In the second half we are the manufacturer, trying to fabricate a user friendly, and fully functioning microfluidic device. As the end user, we have three different T-junction droplet generator devices, with similar geometries. We start investigating by generating water droplets in an oil medium. They self-organise into various flow patterns: single-profile, double-helix profile and triple-helix profile. We document how, with increasing flow rate ratio and capillary number, we observe more densely packed droplet flow patterns. The device with the deeper expansion channel provides more space for the droplets and they self-organise the triple-helix pattern in 3-dimension. We then use the same devices to generate droplets for which we can calculate the volume. The fluid flow in a microchannel happens in four different regimes: ballooning, squeezing, dripping and jetting regimes. In single-cell and single-molecule analysis devices, the ability to create droplets on demand and of a certain volume is a desired capability. This can be achieved by understanding and learning how to use the fluid flow characteristics accurately. We experiment with the three different sized microfluidic devices, to measure the droplet volume throughout the squeezing to dripping regimes. This is achieved by manipulating the capillary number and the flow rate ratio. We observe a similar result as with the flow patterns: that the capillary number has an impact on the droplet volume. As the capillary number increases the droplet diameter decreases. Further, for a set capillary number we can fine tune the droplet diameter by changing the flow rate ratio. As the flow rate ratio increases the volume of water droplets increases, despite the fact the capillary number is set. These coincide with our flow pattern results. Our results fit to the scaling law to predict the droplet size introduced by Tanet al. in 2008 [51]. Unlike some other authors in the literature, we did not observe a critical capillary number where the droplet volume changes suddenly. However, we did observe a transition area where we cannot define the regime of the fluid flow. As the manufacturer we designed and fabricated our own planar free standing microfluidic devices using a polymer called SU-8. After looking into the weaknesses and the strengths of using SU-8, we describe how we successfully fabricated working devices and developeda new procedure in adhesive low temperature bonding. We finish by considering the challenges of connecting micro sized structures to a macro sized syringe pump, and fabricated a chip-holder inspired by applications in industry.

Polymeric microchips have received increasing attention in chemical analysis because polymers have attractive properties, such as low cost, ease of fabrication, biocompatibility and high flexibility. However, commercial polymers usually exhibit analyte adsorption on their surfaces, which can interfere with microfluidic transport in, for example, chemical separations such as chromatography or electrophoresis. Usually, surface modification is required to eliminate this problem. To perform stable and durable surface modification, a new polymer, poly(methyl methacrylate-co-glycidyl methacrylate) (PGMAMMA) was prepared for microchip fabrication, which provides epoxy groups on the surface. Whole surface atom transfer radical polymerization (ATRP) and in-channel ATRP approaches were employed to create uniform and dense poly(ethylene glycol) (PEG)-functionalized polymer brush channel surfaces for capillary electrophoresis (CE) separation of biomolecules, such as peptides and proteins. In addition, a novel microchip material was developed for bioanalysis, which does not require surface modification, made from a PEG-functionalized copolymer. The fabrication is easy and fast, and the bonding is strong. Microchips fabricated from this material have been applied for CE separation of small molecules, peptides, proteins and enantiomers. Electric field gradient focusing (EFGF) is an attractive technique, which depends on an electric field gradient and a counter-flow to focus, concentrate and separate charged analytes, such as peptides and proteins. I used the PEG-functionalized copolymer to fabricate EFGF substrates. The separation channel was formed in an ionically conductive and protein resistant PEG-functionalized hydrogel, which was cast in a changing cross-sectional cavity in the plastic substrate. The hydrogel shape was designed to create linear or non-linear gradients. These EFGF devices were successfully used for protein focusing, and their performance was optimized. Use of buffers containing small electrolyte ions promoted rapid ion transport in the hydrogel for achieving the designed gradients. A PEG-functionalized monolith was incorporated in the EFGF separation channel to reduce dispersion and improve focusing performance. Improvement in peak capacity was proposed using a bilinear EFGF device. Protein concentration exceeding 10,000-fold was demonstrated using such devices.

Thesis (Ph. D.)--West Virginia University, 2006.
Title from document title page. Document formatted into pages; contains xiv, 131 p. : ill. (some col.). Includes abstract. Includes bibliographical references.

Thesis (Ph. D.)--George Mason University, 2007.
Title from PDF t.p. (viewed Jan. 22, 2008). Thesis director: Rao V. Mulpuri. Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Electrical and Computer Engineering. Vita: p. 159. Includes bibliographical references (p. 145-158). Also available in print.

Thesis (Ph. D.)--Massachusetts Institute of Technology, Dept. of Chemical Engineering, 2010.
Cataloged from PDF version of thesis.
Includes bibliographical references (p. [109]-115).
During most of the twentieth century, direct study of individual polymer molecules was impossible due to their small size. Therefore, polymers were typically studied in bulk solutions, and their behavior and interactions had to be understood through average bulk property measurements. Because the scale of most industrial applications greatly exceeded the size of these molecules, this level of analysis was satisfactory. In the last twenty years, however, the appearance of microfluidic devices, whose smallest length scales are comparable to the size of a polymer molecule, has offered ways to visually study the behavior of individual polymer molecules and made possible new and exciting applications that exploit the precise control afforded by the small size of these devices. One such application is gene mapping, which extracts, at a. coarse level, the information embedded in the base pair sequence of genomic DNA. This technology relies on the ability to manipulate single DNA molecules in order to perform such tasks as separating DNA based on length and stretching DNA away from its entropically coiled equilibrium state. Recently, many novel methods have been proposed to accomplish these tasks using microfabricated devices, and munch experimental work has been focused on identifying and characterizing the underlying physics governing these devices. Current understanding, however, is greatly hampered by the fact that experiments can only provide limited information about the behavior of DNA molecules (e.g., they are unable to resolve details on small time and length scales). Therefore, simulations are an invaluable tool in the study of DNA behavior in microfiuidic devices by complementing and guiding experimental investigations. In this thesis, we present Brownian Dynamics simulations of the single molecule behavior of DNA in microfluidic devices related to gene mapping. In particular, we have considered the use of a post array to "precondition" the configuration of molecules for subsequent stretching in a contraction and compared our results to previous experiments. We found good qualitative agreement between experiments and simulations for DNA behavior in the post array, but our simulations consistently overpredicted the final stretch of molecules at the end of the contraction, which we attributed to nonlinear electrokinetic effects. We also investigate the electrophoretic collision of a DNA molecule with a. large, ideally conducting post. Field-induced compression was shown to play a critical role in the escape process of a molecule trapped on the post surface, and an extensive theoretical analysis is performed, describing both the local field-induced compression and the larger collision problem. Finally, we study the relaxation process of an initially stretched molecule in slit-like confinement. We present the first simulation results that exhibit two distinct relaxation times in the linear force regime, as previously reported in recent experiments. Our analysis is focused on the experimentally inaccessible dynamics in the transverse directions, particularly at short times and on small length scales. Comparisons to the predictions of a recent mechanistic model of confined relaxation were found to be satisfactory.
by Daniel Warner Trahan.
Ph.D.

Master of Science
Department of Chemistry
Christopher T. Culbertson
Chemical separation involves selective movement of a component out of a region shared by multiple components into a region where it is the major occupant. The history of the field of chemical separations as a concept can be dated back to ancient times when people started improving the quality of life by separation of good materials from bad ones. Since then the field of chemical separation has become one of the most continually evolving branches of chemical science and encompasses numerous different techniques and principles. An analytical chemist’s quest for a better way of selective identification and quantification of a component by separating it from its mixture is the cause for these ever evolving techniques. As a result, today there are numerous varieties of analytical techniques for the separation of complex mixtures. High Performance Liquid Chromatography (HPLC), Gas Chromatography (GC), Capillary Electrophoresis (CE) and Gel Electrophoresis are a few out of a long list. Each these techniques manipulates the different physical and chemical properties of an analyte to achieve a useful separation and thus certain techniques will be suited for certain molecules. This work primarily focuses on the use of Capillary Electrophoresis as a separation technique. The mechanism of separation in Capillary Zone Electrophoresis and principles of UV detection will discussed in chapter one. Chapter two contains a discussion about the application of Capillary Electrophoresis (CE) on microfluidc devices. This will include sections on: microfabrication techniques of PDMS and photosensitized PDMS (photoPDMS), a UV detector for microfluidic devices and its application for the detection of wheat proteins. In Chapter three we report the experimental part of this project which includes; investigations on the effect of UV exposure time and thermal curing time on feature dimensions of photoPDMS microfluidic device, investigations on the injection and separation performances of the device, characterization of a UV detector set up and its application for the separation and detection of wheat gliadin proteins. The results of these investigations are presented in chapter four.

This work presents the development of 3D printed microfluidic devices and their application to microchip analysis. Initial work was focused on the development of the printer resin as well as the development of the general rules for resolution that can be achieved with stereolithographic 3D printing. The next stage of this work involved the characterization of the printer with a variety of interior and exterior resolution features. I found that the minimum positive and negative feature sizes were about 20 μm in either case. Additionally, micropillar arrays were printed with pillar diameters as small as 16 μm. To demonstrate one possible application of these small resolution features I created microfluidic bead traps capable of capturing 25 μm polystyrene particles as a step toward capturing cells. A second application which I pioneered was the creation of devices for microchip electrophoresis. I separated 3 preterm birth biomarkers with good resolution (2.1) and efficiency (3600 plates), comparable to what has been achieved in conventionally fabricated devices. Lastly, I have applied some of the unique capabilities of our 3D printer to a variety of other device applications through collaborative projects. I have created microchips with a natural masking monolith polymerization window, spiral electrodes for capacitively coupled contactless conductivity detection, and a removable electrode insert chip. This work demonstrates the ability to 3D print microfluidic structures and their application to a variety of analyses.

>Magister Scientiae - MSc
Microfluidic paper based electrochemical sensing devices (μPEDs) provides a new way for point of care testing (POCT). μPEDs offer an inexpensive, portable, easy to use technology too monitor the environment and diagnose diseases, especially in developing countries in cases where there is not enough infrastructure and a limited trained medical and health professionals. The aim of this work is to develop a paper based electrode which can be further integrated into a microfluidic paper device to develop miniature point of care devices. Paper was used as a substrate for printing of the electrode because it is found everywhere, inexpensive and it is compatible with a number of chemical, biochemical and medical applications. Polyamic acid (PAA) was incorporated into commercial carbon ink and was used to print the working electrode. The first part of the study was conducted using the commercial screen printed carbon electrodes (SPCE) to study and understand the electrochemical behaviour of PAA. Cobalt nanoparticles and cobalt nanoparticles‐polyamic acid composites were electrochemically deposited onto SPCE. The modified electrodes were characterised using cyclic voltammetry. As synthesised polyamic acid were characterised using Scanning Electron Microscopy (SEM) to evaluate the morphology and chemical composition of polyamic acid. Transmission Electron Microscopy (TEM) was used to study the particle size and chemical composition of cobalt nanoparticles. Fourier Transform Infrared Spectroscopy (FTIR) was used to study the chemical nature of polyamic acid and cyclic voltammetry (CV) was used to study the electrochemical behaviour of polyamic acid and cobalt nanoparticle electrodes. The diffusion coefficients and formal potential of the electrodes were calculated. The modified and bare electrodes were also used to electrochemically detect Norfloxacin in an aqueous solution by CV and square wave voltammetry (SWV) and the analytical performance of the electrochemical systems are reported here. The obtained limit of detection for the bare SPCE was 3.7 x 10‐3 M and 14.7 x 10‐3 M for the PAA‐SPCE.

Lab-on-a-chip devices, also known as micro total analysis systems (μTAS), are implementations of chemical analysis systems on microchips. These systems can be fabricated using standard thin film processing techniques. Microfluidic and nanofluidic channels are fabricated in this work through sacrificial etching. Microchannels are fabricated utilizing cores made from AZ3330 and SU8 photoresist. Multi-channel electroosmotic (EO) pumps are evaluated and the accompanying channel zeta potentials are calculated. Capillary flow is studied as an effective filling mechanism for nanochannels. Experimental departure from the Washburn model is considered, where capillary flow rates lie within 10% to 70% of theoretical values. Nanochannels are fabricated utilizing cores made from aluminum, germanium, and chromium. Nanochannels are made with 5 μm thick top layers of oxide to prevent dynamic channel deformation. Nanochannel separation schemes are considered, including Ogston sieving, entropic trapping, reptation, electrostatic sieving, and immutable trapping. Immutable trapping is studied through dual-segment nanochannels that capture analytes that are too large to pass from one channel into a second, smaller channel. Polymer nanoparticles, Herpes simplex virus type 1 capsids, and hepatitis B virus capsids are trapped and detected. The signal-to-noise ratio of the fluorescently-detected signal is shown to be greater than 3 for all analyte concentrations considered.

The lymphatic system has three primary roles: transporting lipids, transporting immune cells, and maintaining fluid balance. Each one of these roles are influenced by the presence of flow. Inflammation increases lymph flow, lipid uptake is enhanced by flow, cancer cell migration increases in the presence of flow, and lymphatic permeability and lymphatic contractility respond to changes in flow. Flow is very important to lymphatic function, and yet, there are no in vitro models that incorporate both luminal (flow along cell lumen) and transmural (flow through cell lumina) flow for lymphatics. To address this need, a microfluidic device has been developed that can incorporate both of these types of flow. This is achieved by driving flow through a channel which creates a pressure gradient that drives fluid through a porous membrane into an adjacent channel. Following several design iterations, the device can be easily fabricated, imaged, and cells can grow and survive in it. Permeability experiments have been performed in static and flow, 0.175 mL/min (0.5 dyne/cm²), cases. The effective permeability of dextran in the static and flow cases was calculated to be 0.0083 μm/s and 2.05 μm/s respectively. While the effective permeability of bodipy in the static and flow cases was calculated to be 0.0053 μm/s and 2.57 μm/s respectively. The static values are similar to values obtained in a transwell study by Dixon et al. As mentioned, lipid uptake is increased in the presence of flow and these numbers suggest the same. In addition to permeability studies, experiments were performed with cancer cells suspended in a collagen gel. Two image processing techniques were used to quantify cancer cell migration. The first technique was used to calculate the number of cells present at the beginning of the experiment and the number of cells that were ever present during the experiment in that particular z slice. The static case yielded a cell flux of 15 additional cells. While the two flow cases, within interstitial flow range, had a flux of 24 and 40 cells. This suggests that flow increases migration in cancer cells and is in agreement with the literature. The second technique was used to show that the cells in the static and flow cases are similarly motile, but the flow case is more directed in the z direction towards the membrane. The future work for this device is quite extensive, but a strong foundation centered around basic capabilities like inducing flow, seeding cells, and imaging has been formed.

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2008.
Includes bibliographical references (leaves 140-142).
In this document I discuss the fabrication of metallic, aluminum and aluminum oxide, 3D micro channels, made with standard milling technology, along with two channel closing methods for openable devices: half cured-glued PDMS and Pressure Sensitive Adhesive (PSA) Film. Using the aluminum oxide coated micro channels, along with the half cured-glued PDMS process to close the channels and external fast speed valves for actuation, a microfluidic switch for cell sorting capable of operating at 48 Hz was designed, fabricated and tested. The use of aluminum as a channel substrate provides channel strength and short heat dissipation times, and the use of aluminum oxide enhances light energy absorption, which provides the possibility of further laser actuation. Also, the combination of micro fabrication process and actuation technique makes possible the further scaling and handling of large cells as cardiocytes.
by Marco Aurelio Cartas Ayala.
S.M.

Fluorescence lifetime imaging microscopy (FLIM) is a powerful technique to non-invasively map the physical and chemical environment within microfluidic devices. In this work FLIM has been used in conjunction with a variety of other techniques to provide a greater insight into flow behaviour and fluid properties at the microscale. The pH-sensitive fluorescent dyes, fluorescein and C-SNARF 1, have been used to generate pH maps of microfluidic devices with a time-gated camera and a time-and-space-correlated single photon counting (TSCSPC) detector, respectively. Using time-gated detection and fluorescein, the fluorescence lifetime images allow for direct reading of the pH. The relative contribution to fluorescence of the acid and basic forms of C-SNARF 1 was spatially resolved on the basis of pre-exponential factors, giving quantitative mapping of the pH in the microfluidic device. Three dimensional maps of solvent composition have been generated using 2-photon excitation FLIM (2PE-FLIM) in order to observe the importance of gravitational effects in microfluidic devices. Two fluidic systems have been studied: glycerol concentration in the microfluidic device was measured using Kiton red; water concentration in a methanolic solution was measured using ANS. The density mismatch between two solutions of different composition induced a rotation of the interface between two streams travelling side by side in a microchannel. The experiment has provided evidence of non-negligible gravitational effects in microflows. 2PE-FLIM has superior capability than methods used previously to assess similar phenomena. FLIM and micro-particle imaging velocimetry (μ-PIV) have been implemented on a custom-built open frame microscope and used simultaneously for multimodal mapping of fluid properties and flow characteristics. It has been shown that viscosity mismatch between two streams induces a non-constant advective transport across the channel and results in a flow profile that deviates from the usual Poiseuille profile, characteristic of pressure driven flow in microfluidic devices.

This project investigates the use of surface acoustic waves (SAWs) for applications in low cost, low voltage, digital microfluidic systems. To be able to produce surface acoustic waves, the substrate of the microfluidic device needs to be a piezoelectric material. This study explored the use of two different substrates: 128° Y-cut lithium Niobate (LiNbO₃) and RF magnetron sputtered Zinc Oxide(ZnO) on Silicon (Si) (100). The SAW device incorporates aluminium InterDigital Transducers (IDTs) on LiNbO₃ and ZnO/Si piezoelectric material that acts as an excitation agent to create a surface wave on the substrate. When the signal through the IDT matches the correct frequency, a mechanical wave propagates away from the IDT on the substrate surface. Droplet mixing and movement experiments demonstrate a linear relationship between the applied voltage and droplet movement. Other factors tested are the surface treatment effect on droplet movement and surface temperature effects caused by the SAW mechanical wave. Before droplets could be moved a hydrophobic coating had to be deposited on the surface. The surface coating utilizes the octadecytrichlorosilane (OTS) for both its chemical inertness and bio-compatibility. The OTS coating is smooth and thin and does not effect the propagation of the SAW. The propagation mode of the acoustic wave is determined by the structure of the SAW devices and materials. A higher order harmonic mode wave appears in addition to the fundamental Rayleigh wave for LiNbO₃ samples.  The Rayleigh mode and higher mode- Sezawa mode can be induced for the ZnO/Si SAW devices. These different wave modes have been utilized to induce streaming and manipulate liquid droplets for microfluidic application.

Thesis (S.M.)--Massachusetts Institute of Technology, Dept. of Mechanical Engineering, 2007.
Includes bibliographical references (p. 101-108).
We are developing elastomeric polydimethylsiloxane (PDMS) microfluidic devices incorporated with photoactive thin films to create an implantable artificial respiration platform. Whereas state-of-the-art respiration support machines deliver oxygen gas directly to the blood via external macroscale devices, our technique utilizes a biomimetic photocatalytic process to generate energy from light and thus produce dissolved oxygen from water which is already present in the blood. Blood oxygenation will be achieved by the interaction between the photoactivated metal oxide film and blood in the setting of a molded microfluidic conduit, providing a stable and implantable oxygenation platform. As a basic, scalable building block, we developed a noble "network" design which was structurally similar to the native pulmonary capillary network. The interconnected channel geometry was designed in such a way to minimize shear stress and reduce hemolysis and thrombosis inside the microchannel. It allowed alternative flow pathways in the event of single channel occlusion while minimizing the establishment of detrimental pressure gradients. The hemocompatibility analysis demonstrated that the network construct showed acceptable levels of hemolysis rate (< 8%) and thrombus formation.
(cont.) Critical to the success of this project is the understanding of the manufacture parameters for microfluidic devices molded from elastomeric materials like PDMS. In the initial development of our work, we performed the following three tasks to generate manufacture protocols for elastomeric microfluidic devices that will be ultimately used for biological applications: 1) Curing schedules of the heat-cure PDMS elastomers under various fabrication parameters were characterized. 2) The interlayer bonding chemistry of the double layer PDMS device was analyzed followed by subsequent mechanical analysis. 3) The efficacy of various surface treatment techniques on hydrophobic PDMS surfaces was investigated using fluorescently tagged bacteria (E. Coli) flowed through microchannels as reporter particles to measure non-specific adhesion, which will provide useful information in minimizing channel fouling for biological applications.
by Hyesung Park.
S.M.

Thesis: S.B., Massachusetts Institute of Technology, Department of Mechanical Engineering, 2015.
Cataloged from PDF version of thesis.
Includes bibliographical references (pages 91-92).
Upon examining current methods for printing and patterning of live biological cells, there is a need for a method capable of printing with the resolution of single biological cells to organize them into complex structures. In order to fill this need, building upon previous design of a dynamic lithography system, a stop flow lithography system was implemented capable of patterning individual particles with a mean accuracy of 11.92 [mu]m and a standard deviation of 4.63 [mu]m. This was achieved by improving the tracking capability of the software by measuring the exposure vs velocity relationship to anchor the particle as well as implementing a stop flow lithography based software approach. With the goal of producing 3D functionalized tissue, a 3D printing module was constructed for the dynamic lithography system that constructed microscale parts with a minimum layer height 16.42 [mu]m of and planar resolution of 10 [mu]m, comparable to the top available micro-scale 3D printers. To push the capability of the system, I analyzed and presented the limitations of the process via an opto-thermal model, and a computational fluid dynamics model which is then studied through a previously developed throughput analysis to get a theoretical maximum output of the system. In analyzing the limitations of the printing process, maximum input optical system power was characterized, and a theoretical maximum system throughput of 10,000 particles per second was calculated. This work is a step towards voxel based multimaterial printing, leading to printing of living artificial biological organs, better organs on a chip, or even bionic implants that combine electrical and biological elements.
by Nathan Spielberg.
S.B.

Master of Science
Department of Chemistry
Christopher T. Culbertson
Chemists have been trying to relate the structure and composition of different cereal proteins to their physical properties to better inform their product use for more than 250 years now. Among these cereals, wheat is considered the most important due to its unique ability to form viscoelastic dough and retain gas during fermentation, the latter being important for bread making. This property is due to the endosperm part of wheat that contains proteins mostly gliadins and glutens. It is known that the composition and relative ratio of these proteins is determined by both the growing environment and genetics. Manipulation of the genetics allows one for control of only about 50% of the end use quality of the wheat and the rest is controlled by environment. Currently, the bread making quality of wheat is determined by baking test loaves of bread. This process is time consuming and wasteful. The main goal of this project was to create fingerprints of gliadin proteins for different wheat cultivars as a function of environmental conditions. This would then allow wheat kernels to be analyzed and assessed right after harvest to determine their appropriateness for making the various wheat products. Researchers have tried to create a catalogue of information for individual wheat cultivars by ‘fingerprinting’ the gliadins proteins in wheat using various analytical techniques including capillary electrophoresis (CE). CE offers advantages like high separation efficiency, and faster analysis. Further miniaturization of CE on microfluidic devices has enhanced the speed and efficiency of separation. Furthermore, it is possible to integrate multiple chemical analysis processes like sample preparation, separation and detection in a single microfluidics device. Microfluidic uses micron sized separation channels defined in a glass, quartz or polymer. This dissertation is focused on fabricating multilayer microfluidic devices from Poly(dimethylsiloxane) (PDMS) and using these devices to electrophoretically separate wheat gliadin proteins followed by detection using UV absorption in less than 5 min. PDMS is cheap, easy to fabricate and is optically transparent above ~230nm. Initial results of the UV absorbance detector developed for this device are presented.

Tese (doutorado) - Universidade Federal de Santa Catarina, Centro Tecnológico, Programa de Pós-Graduação em Engenharia Química, Florianópolis, 2016.
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Abstract : In this work, answers to some gaps found in the literature on the field of process intensification in microfluidic devices were proposed. The behavior of a carbon-based composite photocatalyst, specifically a composite of TiO2 and graphene, immobilized on the inner walls of a microchannel reactor, was evaluated and compared with a system containing pristine TiO2. Additionally, a comprehensive computational simulation was performed, based on fundamental physics of semiconductors and considering the coupling of radiation distribution, fluid flow, mass transport and chemical reactions. Moreover, a numerical study was carried out aiming to determine optimal photocatalytic film thicknesses for different illumination mechanisms (backside illumination, BSI, and front-side illumination, FSI) as a function of relevant operational variables and parameters, namely the incident irradiation, the apparent first-order reaction constant, the effective diffusivity and the absorption coefficient. Finally, the possibility of numerically predict the effect of wall wettability on gas-liquid flow pattern developed in microfluidic devices was investigated.

Dispositivos microfluídicos são baseados em microcanais nos quais o diâmetro efetivo é da ordem de centenas de micrômetros, resultando em elevada razão área/volume. Embora um considerável avanço tenha sido observado nessa área nas últimas décadas, resultando, inclusive, em aplicações industriais comercialmente disponíveis, ainda há importantes questões em aberto. Neste trabalho, respostas a algumas dessas questões foram propostas. Em particular, procurou-se determinar o comportamento de dispositivos microfluídicos aplicados à intensificação de processos fotocatalíticos considerando um fotocatalisador compósito (especificamente um compósito de dióxido de titânio e grafeno) imobilizado nas paredes internas. Tal sistema foi, então, comparado a um equivalente no qual dióxido de titânio puro foi imobilizado. As partículas de dióxido de titânio e do compósito de dióxido de titânio-grafeno foram depositadas por meio de um método térmico. Suspensões de TiO2 e TiO2- grafeno foram preparadas e injetadas ao longo de microcanais de chips microfluídicos comerciais construídos com vidro borossilicato. Os dispositivos foram, então, tratados termicamente para promover a evaporação do solvente (água) e a deposição do fotocatalisador nas paredes internas. O processo foi realizado ciclicamente para promover a formação de múltiplas camadas. A evolução da deposição foi avaliada pelo monitoramento dos perfis óticos dos sistemas. Azul de metileno foi usado como reagente modelo em ensaios de fotodegradação. Ensaios preliminares permitiram determinar o efeito dos fenômenos de adsorção e fotólise sobre o comportamento global. Nos experimentos de reação fotocatalisada observou-se que uma maior velocidade de reação inicial foi obtida no microrreator contendo fotocatalisador composto (TiO2-GR) imobilizado nas paredes internas, mas ambos os sistemas (TiO2 e TiO2- GR) exibiram velocidades de reação similares quando o estado estacionário foi alcançado. Verificou-se que a taxa de descolorização do azul de metileno no chip microfluídico foi, aproximadamente, uma ordem de magnitude maior que aquela reportada em sistemas macroscópicos equivalentes em condições experimentais similares. Além disso, investigou-se, neste trabalho, a possibilidade de avaliar teoricamente o comportamento de sistemas microfluídicos aplicados a processos fotocatalíticos com base na física fundamental de semicondutores, bem como a possibilidade de modelar computacionalmente os fenômenos acoplados (distribuição de intensidade luminosa, escoamento, transporte de massa e reação química) que ocorrem em reatores de microcanais (provendo uma estimativa para o desempenho do reator, dos pontos de vista global e local). O modelo computacional foi validado com os resultados experimentais. Na sequência, o modelo computacional foi aplicado para a predição da melhor espessura para o filme fotocatalítico imobilizado nas paredes internas de dispositivos microfluídicos em diferentes condições de iluminação (backside illumination, BSI, e front- side illumination, FSI) como função de variáveis operacionais e parâmetros relevantes, nomeadamente a irradiação incidente, a constante de velocidade de reação aparente de pseudo-primeira ordem, a difusividade efetiva e o coeficiente de absorção do fotocatalisador. Finalmente, a possibilidade de predizer numericamente o efeito da molhabilidade da parede sobre padrões de escoamento multifásicos desenvolvidos em microcanais foi avaliada. Tal modelo computacional pode ser utilizado como fonte de informação prévia sobre o impacto de diferentes propriedades do filme fotocatalítico na morfologia interfacial de escoamento gás-líquido em microrreatores fotoquímicos. Em particular, escoamentos gás-líquido isotérmicos (Taylor e estratificado) foram avaliados através do modelo volume of fluid (VOF). Microcanais com condições limites de hidrofilicidade e hidrofobicidade foram investigados tomando-se como base um referencial experimental disponível na literatura. Um estudo preliminar detalhado foi conduzido para a determinação da malha computacional ótima, capaz de permitir modelagem adequada do filme líquido formado entre as cavidades de gás e a parede sólida, no caso de Taylor flow. Os resultados numéricos foram comparados com dados experimentais (comprimento máximo de cavidade e área de cavidade, para o caso de Taylor flow, e espessura do filme gasoso no caso de escoamento estratificado) e algumas correlações disponíveis (comprimento máximo de cavidade e perda de carga por cavidade) e boa concordância foi observada. Nas mesmas condições de alimentação, o modelo foi capaz de captar os diferentes padrões de escoamento gás-líquido esperados quando o ângulo de contato da parede foi variado. Portanto, tal modelo computacional pode ser utilizado em estudos de scale out com o objetivo de projetar e otimizar reatores compactos modulares baseados na tecnologia de microcanais nos quais escoamento multifásico, particularmente gás e líquido, é estabelecido. Discussões acerca das limitações e de propostas futuras referentes ao desenvolvimento deste trabalho também são apresentadas.

This thesis work concerns the investigation of materials and methods that can be applied to the realization of microfluidic devices (MFDs). In particular, the attention is placed on modular MFDs, as opposed to fully integrated ones. The reasons behind this choice are given in detail in Section 1.2 of this work, but they can be here summarized in the fact that while integrated MFDs offer great advantages in terms of portability, modular devices are more versatile, and so particularly well suited for research applications. The first part of the work here reported describes the microfabrication techniques employed for the realization of single-function microfluidic modules. Devices have been fabricated through PDMS replica molding from SU-8 masters. Masters have been in turn realized through masked UV-lithography or one- or two-photon direct laser writing, depending on the resolution requirements. The replica molding method is a very fast and efficient way to realize MFDs, but suffers from some limitations in the structure shapes that can be successfully replicated. In light of this, a photopolymerizable hybrid organic/inorganic sol-gel blend is proposed and tested as alternative material for MFDs fabrication. The characterization results reveal that this material is biocompatible and features better mechanical properties than PDMS, but structures with more than one dimension exceeding a few micrometers tend to crack during fabrication, making this blend unusable as bulk material. Still, this material could be efficiently employed to fabricate sub-structuration inside PDMS channels. Following this investigation on materials, a microfluidic mixing module is proposed and tested. Since laminar flow conditions dominate inside microchannels, efficient mixing in MFDs require the use of specifically designed mixers. The proposed module makes use of obstructions inside a microchannel to perturb the laminar flow and thus enhance mixing of two species. The most efficient geometries have been selected with the aid of numerical simulations, and two promising layouts have been fabricated and experimentally tested by measuring the dilution of a fluorophore (mixing between a fluorophore solution and pure solvent) through confocal fluorescence microscopy. Thirdly, the fabrication and characterization of an optofluidic light switching module is reported. This device employs a water/air segmented flow generated by a T-junction to alternatively transmit or total-reflect a laser beam. This deflection is proved to be periodical, and its frequency can be varied nonlinearly by adjusting the injection flow rates of air and water. The duty cycle of the module is also characterized, and a method to modulate it by increasing the water temperature is proposed and verified. Finally, a number of attempts to generate a nanoporous, low refractive index PDMS are described. The identification of an efficient procedure to fabricate this kind of material would lead to the possibility of using common microfluidic channels as water-core waveguides. To date, these attempts have not been totally successful, but critical points are identified, and viable strategies for future works on the subject are proposed.
Questo lavoro di tesi tratta dello studio di materiali e metodi che possono essere applicati alla realizzazione di dispositivi microfluidici (DMF). In particolare l’attenzione è rivolta ai dispositivi modulari, piuttosto che a quelli altamente integrati. Le ragioni dietro questa scelta sono spiegate in dettaglio nella Sezione 1.2 di questa tesi, ma possono essere qui sintetizzate nel fatto che anche se i DMF integrati offrono grandi vantaggi in termini di dimensioni finali, i dispositivi modulari sono più versatili, e quindi particolarmente utili per applicazioni nel campo della ricerca. La prima parte del lavoro qui riportato descrive le tecniche di microfabbricazione utilizzate per la realizzazione di moduli microfluidici monofunzionali. I dispositivi sono stati realizzati per replica molding in PDMS a partire da master in SU-8. I master sono stati a loro volta fabbricati tramite litografia UV con maschera oppure per scrittura laser diretta ad uno o due fotoni, a seconda dei requisiti di risoluzione. Il replica molding è un metodo molto rapido ed efficiente per realizzare DMF, ma presenta alcuni limiti per quanto riguarda la forma delle strutture che è possibile replicare con successo. Alla luce di questo, un sol-gel fotopolimerizzabile ibrido organico/inorganico viene qui proposto e testato come materiale alternativo per la fabbricazione di DMF. I risultati della caratterizzazione rivelano che questo materiale è biocompatibile e presenta proprietà meccaniche migliori di quelle del PDMS, ma strutture con più di una dimensione eccedente i pochi micrometri tendono a sviluppare cricche, cosa che impedisce l’utilizzo di questo sol-gel come materiale massivo. Ciononostante, questo sol-gel potrebbe venir efficacemente impiegato per la realizzazione di sottostrutturazioni all’interno di canali microfluidici. Dopo questo studio sui materiali, un modulo microfluidico per il mescolamento è proposto e testato. Dato che le condizioni di flusso laminare sono dominanti all’interno dei microcanali, per ottenere un mescolamento efficiente in un DMF è necessario includere nel dispositivo un miscelatore specificatamente progettato. Il modulo proposto utilizza delle ostruzioni all’interno del microcanale per perturbare il flusso laminare e quindi favorire il mescolamento. Con l’aiuto di alcune simulazioni numeriche, le geometrie più efficienti sono state individuate, e due layout particolarmente promettenti sono stati realizzati e caratterizzati sperimentalmente misurando la diluizione di un fluoroforo (mescolamento tra una soluzione del fluoroforo e puro solvente) attraverso la microscopia confocale di fluorescenza. A seguire, viene riportata la fabbricazione e caratterizzazione di un modulo optofluidico per la deflessione della luce. Questo dispositivo utilizza un flusso segmentato acqua/aria generato da una giunzione a T per trasmettere o riflettere (per riflessione totale interna) alternativamente un fascio laser. Questa alternanza è periodica, e la sua frequenza può essere controllata variando la portata dei flussi iniettati di aria e acqua. Inoltre, il duty cycle del modulo è stato caratterizzato, e viene proposto e verificato un metodo per modularlo attraverso un aumento della temperatura dell’acqua. Infine, vengono descritti alcuni tentativi di generare un PDMS nanoporoso con basso indice di rifrazione. La messa a punto di una procedura efficiente per la fabbricazione di questo genere di materiale porterebbe alla possibilità di usare i classici canali microfluidici come guide d’onda. Al momento questi tentativi hanno avuto solo parziale successo, ma i maggiori punti di criticità sono stati identificati, e vengono proposte alcune strategie per il loro futuro superamento.

The goal of this study is to design and manufacture a microfluidic device capable of measuring changes in impedance valuesof microfluidic cell cultures. Tocharacterize this, an interdigitated array of electrodes was patterned over glass, where it was then bonded to a series of fluidic networks created in PDMS via soft lithography. The device measured ethanol impedance initially to show that values remain consistent over time. Impedance values of water and 1% wt. saltwater were compared to show that the device is able to detect changes in impedance, with up to a 60% reduction in electrical impedance in saltwater. Cells were introduced into the device, where changes in impedance were seen across multiple frequencies, indicating that the device is capable of detecting the presence of biologic elements within a system. Cell measurements were performed using NIH-3T3 fibroblasts.

Delivery vehicles that can encapsulate and release active ingredients of pre-determined volumes at the target site on-demand present a challenge in biomedical field. Due to their tunable physiochemical properties and degradation rate, polymeric particles are one of the most extensively employed delivery vehicles. Generally they are fabricated from emulsion templates. Conventional bulk emulsification technique provides little control over the characteristics of droplets generated. Thus the properties of the subsequent particles cannot be controlled. The advance of droplet microfluidics enables the generation and manipulation of designer single, double or higher-order emulsion droplets with customizable structure. These droplets are powerful and versatile templates for fabricating polymeric delivery vehicles with pre-determined properties. Due to the monodispersity of droplet templates by microfluidics, the relationship between size, size distribution, shape, architecture, elastic responses and release kinetics can be systematically studied. These understandings are of key importance for the design and fabrication of the next generation polymeric delivery vehicles with custom-made functions for specific applications. In the present work, we engineer the droplet templates generated from microfluidics to fabricate designer polymeric microparticles as delivery vehicles. We investigate and obtain the relationship between the particle size, size distribution, structure of microparticles and their release kinetics. Moreover, we also identify an innovative route to tune the particle shape that enables the investigation of the relationship between particle shape and release kinetics. We take advantage of the dewetting phenomena driving by interfacial tensions of different liquid phases to vary the droplet shape. We find that the phase-separation-induced shape variation of polymeric composite particles can be engineered by manipulating the kinetic barriers during droplet shape evolution. To predict the performance of our advanced polymer particles in practical applications, for instance, in narrow blood vessels in vivo, we also develop a novel capillary micromechanics technique to characterize the linear and non-linear elastic response of our polymer particles on single particle level. The knowledge of the mechanical properties enables the prediction as well as the design of the mechanical aspects of polymer particles in different applications. The ability to control and design the physical, chemical, mechanical properties of the delivery vehicles, and the understanding between these properties and the biological functionalities of delivery vehicles, such as the release kinetics, lead towards tailor-designed delivery vehicles with finely-designed functionalities for various biomedical applications. Our proposed electro-microfluidic platform potentially enables generation of submicron droplet templates with a narrow size distribution and nanoscaled delivery vehicles with well-controlled properties, leading to a next generation of intracellular delivery vehicles. Microfluidic-based technique has the potential to be scaled up by parallel operation. Therefore, we are well-equipped for the massive production of custom-made droplet templates of both micron-size and nanosized, and we can design the physiochemical properties and biological functionalities of the delivery vehicles. These abilities enable us to provide solutions for applications and fundamental topics where encapsulation, preservation and transportation of active ingredients are needed.
published_or_final_version
Mechanical Engineering
Doctoral
Doctor of Philosophy

In recent years, the LOC community has focused most of its research in the biomedical and biotechnology fields, due to the need of portable, low power consumption and low cost theranostics microdevices. Some developing countries do not have suitable medical diagnostics technologies and the supply and storage of the reagents is in many cases limited as well as the access to energy. Furthermore, developed countries are experimenting population aging needing novel low cost efficient disease-screening technologies. The introduction of LOC and microfluidics allow the integration of complex functions that could lead to the developing of more accurate, cheap and reliable theranostic tools. Current focus of application is focused mostly in drug delivery 1, cellular analysis 2, and disease diagnosis 3. Microfluidics is improving the developing of novel point-of-care devices, but there are some challenges that are slowing down the massive production of these LOC. These areas include new methods for sample collection, world-to-chip interfaces, sample pre-treatment, improvement of long-term stability of reagents, working with complex sample specimens, multiple detection of biomarkers and simplify the read-out 4. The main aim of this thesis work was to create novel, cheap and with a high degree of automatization miniaturized biosensing devices with the objective to facilitate Point-of-Care diagnostics in the near future. Our efforts have been focused into developing a LOC system with electrochemical sensing capabilities adjustable to any biomarker, depending only on sample volumes and required analysis times. The devices integrate low-cost label-free biosensors exploiting microfluidics-based self-functionalization, or specialization. The biosensor functionalization takes place in situ and selectively, just before the sensing, and their area keeps dry and inactive until the test starts. The reagents and the sensing parts are kept separated and brought into contact just before the test, avoiding the need of complex fabrication and storage methods to guarantee functionalization integrity. The novel design reduces the cost of the final instrumentation, by simplifying the measurements, while keeping sensitivities and LODs relevant for the application. Furthermore, since the interaction of antibody and protein is time and concentration dependent, our device has the capability to adjust its sensitivity. We have tuned and characterized our system sensitivity using different biomarkers. The development of our novel devices was possible by exploiting synergies in disciplines previously studied in our group. Particularly, in fields such as microfluidics 5-8, surface functionalization 9-14 and electrochemical biosensors 15-19. Summarizing, we are proposing novel microfluidic devices with integrated biosensors. The systems are based on the principle of laminar co-flow in order to perform an on-chip selective surface bio-functionalization of LOC integrated biosensors. This method has the advantage of performing the surface modification protocols “in situ” before the detection. The system can be easily scaled to incorporate several sensors with different biosensing targets in a single chip. We are proposing a novel voltage and impedance differential measurements; that allow us to simplify the read-out. As biomedical application we focus our attention on the detection of prostate cancer biomarkers. Bibliography 1. I. U. Khan, C. A. Serra, N. Anton and T. Vandamme, Journal of Controlled Release, 2013, 172, 1065-1074. 2. H. Andersson and A. Van den Berg, Sensors and Actuators B: Chemical, 2003, 92, 315-325. 3. M. J. Cima, Annual Review of Chemical and Biomolecular Engineering, 2011, 2, 355-378. 4. C. D. Chin, V. Linder and S. K. Sia, Lab on a Chip, 2012, 12, 2118-2134. 5. R. Rodriguez-Trujillo, C. A. Mills, J. Samitier and G. Gomila, Microfluidics and Nanofluidics, 2007, 3, 171-176. 6. R. Rodriguez-Trujillo, O. Castillo-Fernandez, M. Garrido, M. Arundell, A. Valencia and G. Gomila, Biosensors and Bioelectronics, 2008, 24, 290-296. 7. O. Castillo-Fernandez, R. Rodriguez-Trujillo, G. Gomila and J. Samitier, Microfluidics and Nanofluidics, 2014, 16, 91-99. 8. J. Comelles, V. Hortigüela, J. Samitier and E. Martínez, Langmuir, 2012, 28, 13688-13697. 9. E. Prats-Alfonso, F. García-Martín, N. Bayo, L. J. Cruz, M. Pla-Roca, J. Samitier, A. Errachid and F. Albericio, Tetrahedron, 2006, 62, 6876-6881. 10. J. Vidic, M. Pla-Roca, J. Grosclaude, M.-A. Persuy, R. Monnerie, D. Caballero, A. Errachid, Y. Hou, N. Jaffrezic-Renault, R. Salesse, E. Pajot-Augy and J. Samitier, Analytical Chemistry, 2007, 79, 3280-3290. 11. Y. Hou, S. Helali, A. Zhang, N. Jaffrezic-Renault, C. Martelet, J. Minic, T. Gorojankina, M.-A. Persuy, E. Pajot-Augy, R. Salesse, F. Bessueille, J. Samitier, A. Errachid, V. Akimov, L. Reggiani, C. Pennetta and E. Alfinito, Biosensors and Bioelectronics, 2006, 21, 1393-1402. 12. S. Rodríguez Seguí, M. Pla, J. Minic, E. Pajot‐Augy, R. Salesse, Y. Hou, N. Jaffrezic‐Renault, C. A. Mills, J. Samitier and A. Errachid, Analytical Letters, 2006, 39, 1735-1745. 13. A. Lagunas, J. Comelles, E. Martínez and J. Samitier, Langmuir, 2010, 26, 14154-14161. 14. A. Lagunas, J. Comelles, S. Oberhansl, V. Hortigüela, E. Martínez and J. Samitier, Nanomedicine: Nanotechnology, Biology and Medicine, 2013, 9, 694-701. 15. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Materials Science and Engineering: C, 2008, 28, 680-685. 16. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Sensors and Actuators B: Chemical, 2007, 120, 615-620. 17. M. Kuphal, C. A. Mills, H. Korri-Youssoufi and J. Samitier, Sensors and Actuators B: Chemical, 2012, 161, 279-284. 18. D. Caballero, E. Martinez, J. Bausells, A. Errachid and J. Samitier, Analytica Chimica Acta, 2012, 720, 43-48. 19. M. Barreiros dos Santos, J. P. Agusil, B. Prieto-Simón, C. Sporer, V. Teixeira and J. Samitier, Biosensors and Bioelectronics, 2013, 45, 174-180.
En años recientes, la comunidad de LOC ha enfocado todos sus esfuerzos en la investigación de nuevas aplicaciones para la biomedicina y biotecnología. Algunos países en vías de desarrollados no tienen tecnologías de diagnóstico adecuadas, además el suministro y almacenamiento de los reactivos es en muchos casos limitado, y en ocasiones cuentan con un acceso limitado al consumo de energía. Por otra parte, los países desarrollados se han encontrado con una población envejecida, y por lo tanto se ha generado la necesidad de contar con nuevas tecnologías para el diagnóstico de enfermedades las cuales sean accesibles y orientadas a una terapia más personalizada. Tanto la microfluídica como los LOC han permitido la integración de funciones de análisis complejas capaces de desarrollar herramientas de diagnostico más precisas, de bajo coste y confiables. Actualmente toda la atención se ha centrado en el diseño de aplicaciones para administración de fármacos 1, análisis celular 2 y diagnostico de enfermedades 3. La introducción de la microfluídica ha servido para mejorar el desarrollo de nuevos dispositivos point-of-care, pero todavía existen algunos problemas que han evitado la producción masiva de estos LOC. Las áreas en las que se pretende conseguir una mejora son la recolección de la muestra, mejora de la interfaz entre el chip y el usuario, tratamiento previo de la muestra, mejorar la estabilidad de los reactivos, trabajo con muestras complejas, detección múltiple de biomarcadores y simplificación del sistema de medida 4. Nuestros esfuerzos se han dedicado en desarrollar un sistema LOC con capacidad de detección electroquímica ajustable a cualquier biomarcador, dependiendo únicamente en la cantidad de muestra y los tiempos de análisis. Nuestros dispositivos microfluídicos cuentan con biosensores integrados de bajo coste con capacidad de auto-funcionalización. La funcionalización de los biosensores se realiza in-situ y selectivamente, antes de la detección, manteniendo el área de detección inerte hasta el inicio de la prueba. Los reactivos y el área de detección se almacenan por separado y entran en contacto hasta el inicio del experimento, lo cual facilita el método de fabricación. Se ha podido desarrollar este trabajo gracias a los estudios previos realizados en nuestro grupo en distintas disciplinas, tales como: microfluídica 5-8, funcionalización de superficies 9-14 y biosensores electroquímicos 15-19. Bibliografía 1. I. U. Khan, C. A. Serra, N. Anton and T. Vandamme, Journal of Controlled Release, 2013, 172, 1065-1074. 2. H. Andersson and A. Van den Berg, Sensors and Actuators B: Chemical, 2003, 92, 315-325. 3. M. J. Cima, Annual Review of Chemical and Biomolecular Engineering, 2011, 2, 355-378. 4. C. D. Chin, V. Linder and S. K. Sia, Lab on a Chip, 2012, 12, 2118-2134. 5. R. Rodriguez-Trujillo, C. A. Mills, J. Samitier and G. Gomila, Microfluidics and Nanofluidics, 2007, 3, 171-176. 6. R. Rodriguez-Trujillo, O. Castillo-Fernandez, M. Garrido, M. Arundell, A. Valencia and G. Gomila, Biosensors and Bioelectronics, 2008, 24, 290-296. 7. O. Castillo-Fernandez, R. Rodriguez-Trujillo, G. Gomila and J. Samitier, Microfluidics and Nanofluidics, 2014, 16, 91-99. 8. J. Comelles, V. Hortigüela, J. Samitier and E. Martínez, Langmuir, 2012, 28, 13688-13697. 9. E. Prats-Alfonso, F. García-Martín, N. Bayo, L. J. Cruz, M. Pla-Roca, J. Samitier, A. Errachid and F. Albericio, Tetrahedron, 2006, 62, 6876-6881. 10. J. Vidic, M. Pla-Roca, J. Grosclaude, M.-A. Persuy, R. Monnerie, D. Caballero, A. Errachid, Y. Hou, N. Jaffrezic-Renault, R. Salesse, E. Pajot-Augy and J. Samitier, Analytical Chemistry, 2007, 79, 3280-3290. 11. Y. Hou, S. Helali, A. Zhang, N. Jaffrezic-Renault, C. Martelet, J. Minic, T. Gorojankina, M.-A. Persuy, E. Pajot-Augy, R. Salesse, F. Bessueille, J. Samitier, A. Errachid, V. Akimov, L. Reggiani, C. Pennetta and E. Alfinito, Biosensors and Bioelectronics, 2006, 21, 1393-1402. 12. S. Rodríguez Seguí, M. Pla, J. Minic, E. Pajot‐Augy, R. Salesse, Y. Hou, N. Jaffrezic‐Renault, C. A. Mills, J. Samitier and A. Errachid, Analytical Letters, 2006, 39, 1735-1745. 13. A. Lagunas, J. Comelles, E. Martínez and J. Samitier, Langmuir, 2010, 26, 14154-14161. 14. A. Lagunas, J. Comelles, S. Oberhansl, V. Hortigüela, E. Martínez and J. Samitier, Nanomedicine: Nanotechnology, Biology and Medicine, 2013, 9, 694-701. 15. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Materials Science and Engineering: C, 2008, 28, 680-685. 16. M. Castellarnau, N. Zine, J. Bausells, C. Madrid, A. Juárez, J. Samitier and A. Errachid, Sensors and Actuators B: Chemical, 2007, 120, 615-620. 17. M. Kuphal, C. A. Mills, H. Korri-Youssoufi and J. Samitier, Sensors and Actuators B: Chemical, 2012, 161, 279-284. 18. D. Caballero, E. Martinez, J. Bausells, A. Errachid and J. Samitier, Analytica Chimica Acta, 2012, 720, 43-48. 19. M. Barreiros dos Santos, J. P. Agusil, B. Prieto-Simón, C. Sporer, V. Teixeira and J. Samitier, Biosensors and Bioelectronics, 2013, 45, 174-180.

This research undertaken involved designing, fabricating and testing of a microfiuidic peR micro device with real-time electrochemical detection. The aim was to provide an analytical device which could lower the cost and the time taken for running DNA amplification. The later addition of automated sample handling and detection would thereby reduce the time taken and consequently the overall cost. The real-time electrochemical detection utilised an electrochemical assay for the detection of DNA invented by Molecular Sensing Ltd. It used a single strand ferrocenylated probe DNA molecule which could be detected with an electrochemical cell. The integration of an electrochemical cell was a key feature of this work, along with the immobilisation of the Taq polymerase at its working temperature. Taq polymerase enzyme and T7 polymerase enzyme were immobilised on to microspheres. Taq polymerase was immobilised in four ways and T7 polymerase was immobilised in only one method. After immobilisation the enzymes were unable to amplify DNA within peR experiments. Microfiuidic peR devices, which incorporate the above two features, were designed and fabricated. 3 basic ideas of devices were investigated, fiowthrough, straight line and cyclic triangle device. All the devices had fundamental problems which inhibited there ability to successfully amplify DNA. An electrochemical assay was used within a microfiuidic device with internal electrochemical detection, which utilised a filter to bring about sequence specific DNA detection. Using biotinylated complementary probe DNA attached to streptavidin coated beads to hybridise to the sample DNA. This device incorporated a solid phase extraction and clean up step as well as producing sequence specific DNA detection.

Controlling the behaviour of atmospheric pressure plasmas and their interaction with polymeric materials is of major interest for surface modification applications across multidisciplinary fields intersecting biomedical engineering, bio-nanoengineering, clinical/medical science, material science and microelectronics. The aim of the present work is to investigate the behaviour of atmospheric pressure dielectric barrier discharges in closed systems (microfluidic devices) and open systems (glass capillary devices) and their polymer-surface interactions. Atmospheric pressure microplasma jets operating in helium gas have been used to modify locally the surface energy of polystyrene (PS) and to interact directly with the surface of analytes using a novel plasma assisted desorption ionisation (PADI) method causing desorption and ionization to occur. Although atmospheric pressure micro-jets are now widely studied for the treatment of materials there is still a lack of understanding of the fundamental plasma-surface processes. A number of recent studies using plasma micro-jets for the surface modification of polymerics have used systems in which the emerging plume impinges directly the substrate head-on. Here, by placing the micro-jet side-on to the substrate we can observe how different flow regions of the jet affect the sample, allowing individual effects to be seen. In addition, this configuration may prove an efficient way of treating samples with reduced or no surface damage. These conclusions are considered to be an important contribution to the study of complex mechanisms underpinning the behaviour of radicals and reactive species in surface modification processes of polymeric materials. The study of the behavioural mechanism involved in the plasma was done using various diagnostic techniques such as electrical measurements, optical emission spectroscopy (OES), Time-averaged and time-resolved ICCD Optical Imaging and Schlieren Photography. The filamentary discharge mode was observed in bonded microchannels using metallic and liquid-patterned electrodes. The treated surfaces were characterised using various techniques such as X-ray photoelectron spectroscopy (XPS), Atomic Force Microscopy (AFM), Optical profiling measurements and Water Contact Angle (WCA) measurements. Schlieren photography has been used to indentify regions of laminar (pre-onset of visual instability) and turbulent flows (post-onset of visual instability) in the exiting gas stream and the nature of their interaction with the substrate surface. The length of both regions varies depending on operating parameters such as frequency, applied voltage and flow rate. WCA results from treated polystyrene (PS) samples exposed directly facing the microjet reveals a change from hydrophobic (high contact angle) to a hydrophilic (low contact angle) surface with substantial reductions in WCA (~ 50 to 60 °) occurring in downstream regions where the turbulent gas mixed with air impinges the substrate surface. In contrast, only small changes in WCA (~ 10 to 20 °) occur in regions where the gas flow is laminar. AFM imaging of treated PS samples reveal holes and ripple like effect with a much larger area than that of the capillary seen on treated samples positioned “head-on” and directly facing the sample but this was not seen using the side-on configuration. The results indicate that excited air species (either mixed or entrained in the He gas flow) which exist only in regions of turbulence are the main agents causing surface covalent bond breaking leading to surface modification. This thesis reports on atmospheric pressure microdischarges and their applications, a brief summary of work done so far including major results, using new and existing technologies including those under development in terms of design, properties and working conditions.