This PDF file contains the front matter associated with SPIE Proceedings Volume 9705, including the Title Page, Copyright information, Table of Contents, Introduction, and the Conference Committee listing.

Efforts to transfer intact mammalian chromosomes between cells have been attempted for more than 50 years with the consistent result being transfer of sub unit length pieces regardless of method. Inertial microfluidics is a new field that has shown much promise in addressing the fractionation of particles in the 2-20 μm size range (with unknown limits) and separations are based upon particles being carried by curving confined flows (within a spiral shaped, often rectangular flow chamber) and migrating to stable “equilibrium” positions of varying distance from a chamber wall depending on the balance of dean and lift forces. We fabricated spiral channels for inertial microfluidic separations using a standard soft lithography process. The concentration of chromosomes, small contaminant DNA and large cell debris in each outlets were evaluated using microscope (60X) and a flow cytometer. Using Dean Flow Fractionation, we were able to focus 4.5 times more chromosomes in outlet 2 compared to outlet 4 where most of the large debris is found. We recover 16% of the chromosomes in outlet #1- 50% in 2, 23% in 3 and 11% in 4. It should be noted that these estimates of recovery do not capture one piece of information- it actually may be that the chromosomes at each outlet are physically different and work needs to be done to verify this potential.

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.

Adaptive elastomer-liquid lens can find a variety of optical applications due to the tunable optical powers without additional lens replacement or displacement. Most current elastomer-liquid lenses use elastomer membrane with a constant thickness. This approach, however, suffers from substantial optical aberration due to the edge clamping effect. In this study, a varied thickness elastomer membrane with customized aspherical profile is designed and developed to encapsulate a plano-convex liquid lens. Such varied thickness membrane is fabricated by double-side replica molding against a deformed elastomer-liquid lens membrane with a constant thickness. Such configuration could alleviate the edge clamping effect. Simulation and experimental results both show that the lens with a varied thickness membrane exhibits improved optical resolutions at both the center and the peripheral regions at the back focal length of 10 mm comparing to the lens with a constant thickness membrane. This study provides an effective solution to suppress the optical aberrations without sacrifice of the optical aperture.

Hydrodynamic 3D flow-focusing techniques in microfluidics are categorized as (a) sheathless techniques which require high flow rates and long channels, resulting in high operating cost and high flow rates which are inappropriate for applications with flow rate limitations, and (b) sheath-flow based techniques which usually require excessive sheath flow rate to achieve hydrodynamic 3D flow-focusing. Many devices based on these principles use complicated fabrication methods to create multi-layer microchannels. We have developed a sheath-flow based microfluidic device that is capable of hydrodynamic 3D self-flow-focusing. In this device the main flow (black ink) in a low speed, and a sheath flow, enter through two inlets and enter a 180 degree curved channel (300 × 300 μm cross-section). Main flow migrates outwards into the sheath-flow due to centrifugal effects and consequently, vertical focusing is achieved at the end of the curved channel. Then, two other sheath flows horizontally confine the main flow to achieve horizontal focusing. Thus, the core flow is three-dimensionally focused at the center of the channel at the downstream. Using centrifugal force for 3D flow-focusing in a single-layer fabricated microchannel has been previously investigated by few groups. However, their demonstrated designs required high flow speed (>1 m/s) which is not suitable for many applications that live biomedical specie are involved. Here, we introduce a new design which is operational in low flow speed (<0.05 m/s) and is suitable for applications involving live cells. This microfluidic device can be used in detecting, counting and isolating cells in many biomedical applications.

Conventional flow control systems, such as syringe and peristaltic pumps, are not well adapted to the control of flow in microchannels. They often result in long equilibration times, hysteresis and low stability. Herein, we present a new method to control the flows in microchannels based on a pressure actuation, by pressurization of reservoirs filled with fluids to be injected in the microsystem. The regulated pressure within the reservoirs generates pulse-free and very stable flows through the microchannels, with short settling time. To control the flow-rate with pressure actuation, highly precise flow sensors are implemented in the fluidic system and an algorithm has been developed to adjust automatically the pressure orders to reach the targeted flow-rates. Unlike a conventional PID regulation which is very sensitive to any transient behavior, our algorithm deals with any coupling effect between the different channels, and is designed to deliver the fastest and the most stable flow response. It calculates a matrix image of the microsystem with the relations between each actuated pressure channels and the measured flow-rates. Furthermore, the system can cope with any external disturbances of the system (presence of air bubble, partial clogging, variation of viscosity or temperature, etc…), and continuously re-adjust the applied pressures. The technology is perfectly suited for droplet manipulation experiments (among other applications) where we can generate 2pL water-in-oil droplets with very high monodispersity (1.63% CV) at an up to 12 kHz/s frequency. Only few seconds are needed to stop the droplets flow, reducing costs by a huge factor.

The academic community knows cyclic olefin copolymer (COC) as a well suited material for microfluidic applications because COC has numerous interesting properties such as high transmittance, good chemical resistance and good biocompatibility. Here we present a fast and cost-effective method for bonding of two COC substrates: exposure to appropriate solvents gives a tacky COC surface which when brought in contact with untreated COC forms a strong and optical clear bond. The bonding process is carried out at room temperature and takes less than three minutes which makes it significantly faster than currently described methods: This method does not require special lab equipment such as hot plates or hydraulic presses. The mild conditions of the bond process also allow for such “tacky COC” lids to be used for sealing of microfluidic chips containing immobilized protein patterns which is of high interest for immunodiagnostic testing inside microfluidic chips.

In this research, electrowetting liquid lens array on curved substrates is developed for wide field of view image sensor. In the conventional image sensing system, this lens array is usually in the form of solid state. However, in this state, the lens array which is similar to insect-like compound eyes in nature has several limitations such as degradation of image quality and narrow field of view because it cannot adjust focal length of lens. For implementation of the more enhanced system, the curved array of lenses based on electrowetting effect is developed in this paper, which can adjust focal length of lens. The fabrication of curved lens array is conducted upon the several steps, including chamber fabrication, electrode & dielectric layer deposition, liquid injection, and encapsulation. As constituent materials, IZO coated convex glass, UV epoxy (NOA 68), DI water, and dodecane are used. The number of lenses on the fabricated panel is 23 by 23 and each lens has 1mm aperture with 1.6mm pitch between adjacent lenses. When the voltage is applied on the device, it is observed that each lens is changed from concave state to convex state. From the unique optical characteristics of curved array of liquid lenses such as controllable focal length and wide field of view, we can expect that it has potential applications in various fields such as medical diagnostics, surveillance systems, and light field photography.

Nanoscale patterned structures composed of biomaterials exhibit great potential for the fabrication of functional biostructures. In this paper, we report cost-effective, rapid, and highly reproducible soft lithographic transfer-molding techniques for creating periodic micro- and nano-scale textures on poly (L-lactic acid) (PLLA) surface. These artificial textures can increase the overall surface area and change the release dynamics of the therapeutic agents coated on it. Specifically, we use the double replication technique in which the master pattern is first transferred to the PDMS mold and the pattern on PDMS is then transferred to the PLLA films through drop-casting as well as nano-imprinting. The ensuing comparison studies reveal that the drop-cast PLLA allows pattern transfer at higher levels of fidelity, enabling the realization of nano-hole and nano-cone arrays with pitch down to ~700 nm. The nano-patterned PLLA film was then coated with rapamycin to make it drug-eluting.

Based on the world-first 3D gel printing technology, we aim to develop 3D gel printing system to realize free-shape design of soft and wet materials. We defined ‘Designable Gels’ as revolutionary gels whose molecular structure, shape, and functions can be designed by users. By virtue of the 3D gel printing system, we can use 3D high-performance gels materials and realize both designed 3D shape and designed properties. At the same time, analysis technology with scanning microscopic light scattering will be immediately used to guarantee the quality of manufactured gels. We believe we will contribute to extend the fields of medical and robot applications and create new markets.

The use of acoustic energy to manipulate particles and cells (acoustophoresis) is a well-studied and popular technique in microfluidic microscopy applications. This powerful and gentle method has typically required the construction of costly and labour intensive resonating chambers. A new fabrication method for acoustophoretic resonators is presented, using inexpensive materials and without the need for cleanrooms or the special equipment typically found within them. By utilizing a simple glass and polyimide sandwiching technique, single, bifurcating, and trifurcating microchannels were built and tested. Various half and full wavelength transversal resonators were established in microchannel widths of 300, 600, and 750 μm using 1, 2.5, and 5 MHz ultrasound. In significantly simplifying the fabrication and prototyping of these microfluidic resonators we hope to address some of the major drawbacks preventing acoustophoresis technology from being incorporated into the toolkits of laboratories around the world.

Microbubbles have been utilized as micro-pumps, micro-mixers, micro-valves, micro-robots and surface cleaners. Various generation techniques can be found in the literature, including resistive heating, hydrodynamic methods, illuminating patterned metal films and noble metal nanoparticles of Au or Ag. We present photothermal microbubble generation by irradiating nanoporous gold disk covered microfluidic channels. The size of the microbubble can be controlled by adjusting the laser power. The dynamics of both bubble growth and shrinkage are studied. The advantages of this technique are flexible bubble generation locations, long bubble lifetimes, no need for light-adsorbing dyes, high controllability over bubble size, low power consumption, etc. This technique has the potential to provide new flow control functions in microfluidic devices.

Traditional bacterial identification methods take one to two days to complete, relying on large bacteria colonies for visual identification. In order to decrease this analysis time in a cost-effective manner, a method to sort and concentrate bacteria based on the bacteria’s characteristics itself is needed. One example of such a method is dielectrophoresis, which has been used by researchers to separate bacteria from sample debris and sort bacteria according to species. This work presents variations in which dielectrophoresis can be performed and their associated drawbacks and benefits specifically to bacterial identification. In addition, a potential microfluidic design will be discussed.

Lab-on-a-chip devices are often applied to point-of-care diagnostic solutions as they are low-cost, compact, disposable, and require only small sample volumes. For such devices, various reagents are required for sample preparation and analysis and, for an integrated solution to be realized, on-chip reagent storage and automated introduction are required. This work describes the implementation and characterization of effective liquid reagent storage and release mechanisms utilizing blister pouches applied to various point-of-care diagnostic device applications. The manufacturing aspects as well as performance parameters are evaluated.

Through further development, integration and validation of micro-nano-bio and biophotonics systems FP7 CanDo is developing an instrument that will permit highly reproducible and reliable identification and concentration determination of rare cells in peripheral blood for two key societal challenges, early and low cost anti-cancer drug efficacy determination and cancer diagnosis/monitoring.

A cellular link between the primary malignant tumour and the peripheral metastases, responsible for 90% of cancerrelated deaths, has been established in the form of circulating tumour cells (CTCs) in peripheral blood. Furthermore, the relatively short survival time of CTCs in peripheral blood means that their detection is indicative of tumour progression thereby providing in addition to a prognostic value an evaluation of therapeutic efficacy and early recognition of tumour progression in theranostics. In cancer patients however blood concentrations are very low (=1 CTC/1E9 cells) and current detection strategies are too insensitive, limiting use to prognosis of only those with advanced metastatic cancer. Similarly, problems occur in therapeutics with anti-cancer drug development leading to lengthy and costly trials often preventing access to market.

The novel cell separation/Raman analysis technologies plus nucleic acid based molecular characterization of the CanDo platform will provide an accurate CTC count with high throughput and high yield meeting both key societal challenges. Being beyond the state of art it will lead to substantial share gains not just in the high end markets of drug discovery and cancer diagnostics but due to modular technologies also in others. Here we present preliminary DNA hybridization sensing results.

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.

Low cost, portable chip based flow cytometry has great potential for applications in resource poor and point of care settings. Typical approaches utilise low cost silicon or glass substrates with light emission and detection performed either off-chip using external equipment or incorporated on-chip using ‘pick and place’ diode lasers and photo-detectors. The former approach adds cost and limits portability while the sub-micron alignment tolerances imposed by the application make the latter impractical for all but the simplest of systems. Use of an optically active semiconductor substrate, on the other hand, overcomes these limitations by allowing multiple laser/detector arrays to be formed in the substrate itself using high resolution lithographic techniques. The capacity for multiple emitters and detectors on a single chip not only enables parallel measurement for increased throughput but also allows multiple measurements to be performed on each cell as it passes through the system. Several different experiments can be performed simultaneously and throughput demand can be reduced with the facility for error checking. Furthermore, the fast switching times inherent with semiconductor lasers allows the active sections of the device to be reconfigured on a sub-microsecond time scale providing additional functionality. This is demonstrated here in a capillary fill system using pairs of laser/detectors that are operated in pulsed mode and alternated between lasing and detecting in an interleaved manner. Passing cells are alternately interrogated from opposing directions providing information that can be used to correct for differences in lateral cell position and ultimately differentiate blood cell type.

This paper details the development of a three dimensional (3D) printing system with a modified microfluidic printhead used for the generation of complex vascular tissue scaffolds. The print-head features an integrated coaxial nozzle that allows the fabrication of hollow, calcium-polymerized alginate tubes that can easily be patterned using 3Dbioprinting techniques. This microfluidic design allows the incorporation of a wide range of scaffold materials as well as biological constituents such as cells, growth factors, and ECM material. With this setup, gel constructs with embedded arrays of hollow channels can be created and used as a potential substitute for blood vessel networks.

Optical micromanipulation techniques and microfluidic techniques can be used in same platform for manipulating biological samples at single cell level. Novel microfluidic devices with integrated channels and waveguides fabricated using ultrafast laser inscription combined with selective chemical etching can be used to enable sorting and isolation of biological cells. In this paper we report the design and fabrication of a three dimensional chip that can be used to manipulate single cells in principle with a higher throughput than is possible using optical tweezers. The capability of ultrafast laser inscription followed by selective chemical etching to fabricate microstructures and waveguides have been utilised to fabricate the device presented in this paper. The complex three dimensional microfluidic structures within the device allow the injected cell population to focus in a hydrodynamic flow. A 1064 nm cw laser source, coupled to the integrated waveguide, is used to exert radiation pressure on the cells to be manipulated. As the cells in the focussed stream flow past the waveguide, optical scattering force induced by the laser beam pushes the cell from out of the focussed stream to the sheath fluid, which can be then collected at the outlet. Thus cells can be controllably deflected from the focussed flow to the side channel for downstream analysis or culture.

A challenging request in the fabrication of microfluidics and biomedical microsystems is a flexible ink-jet printing for breaking the rigidity of classical lithography. A pyroelectric-EHD system is presented. The system has proved challenging spatial resolution down to nanoscale, printing of high ordered patterns, capability of dispensing bio-ink as DNA and protein array for biosensing fabrication, single cells printing and direct printing of nanoparticles. With the method proposed high viscous polymers could be easily printed at high resolution in 2D or in 3D configuration. The pyro-EHD process has been proved for the fabrication of biodegradable microneedles for trasndermal drug delivery and 3D optical waveguides.

Biomedical sensors using microfluidic channels are prone to blockage due to particles and bubbles in the fluid. Wider channels may be used, but wide polymer channels may suffer from structural instability (e.g., sagging channel covers). A common design uses many parallel flow channels separated by structural support walls, but these can be rapidly blocked by particulates. We have been studying an alternative “Cathedral Chamber” design where the channel “roof” (cover) is support by periodic posts which creates many possible flow paths to bypass blockages. We use Monte Carlo modelling with iterative COMSOL fluid dynamics simulations to establish the stream lines, and particle velocities. Then a rules based methodology iteratively places trapped particles based on the fluid paths created by the existing blockages, until the system become fully blocked. Previous work has shown that the periodic post design increases lifetime by allowing 6 to 7 times more blockages than can a parallel channel design. In this paper, we simulate and analyze why expanding the number of channels increases almost linearly the number of particles required for blockages. Lifetime increase is still 4.5-5.5 times even for the limiting case of a 2 channel cathedral chamber. This shows the sideways flow created by the periodic posts creates many advantages for the microfluidic chambers.

This paper reports the maskless fabrication of a microfluidic device with interdigitated electrodes (IDE) based on the technology of MicroElectroMechanical Systems on Printed Circuit Board (PCB-MEMS) and laser ablation. The device has flame retardant (FR)-4 resin as substrate, cooper (Cu) as active material and SU-8 polymer as structural material. By adjusting the laser parameters, Cu IDEs and SU-8 microchannels were successfully patterned onto the FR-4 substrate. The respective width, gap and overlap of the IDEs were 50 μm, 25 μm and 500 μm. The respective width, depth and length of the microchannels were 210 μm, 24.6 μm and 6.3 mm. The resolution and repeatability achieved in this approach, along with the low cost of the involved materials and techniques, enable an affordable micromachining platform with rapid fabrication-test cycle to develop active multiphysic microdevices with several applications in the fields of biosensing, cell culture, drug delivery, transport and sorting of molecules, among others.

We experimentally demonstrate applications of photonic crystal waveguide based devices for on-chip optical absorption spectroscopy for the detection of chemical warfare simulant, triethylphosphate as well as applications with photonic crystal microcavity devices in the detection of biomarkers for pancreatic cancer in patient serum and cadmium metal ions in heavy metal pollution sensing. At mid-infrared wavelengths, we experimentally demonstrate the higher sensitivity of photonic crystal based structures compared to other nanophotonic devices such as strip and slot waveguides with detection down to 10ppm triethylphosphate. We also detected 5ppb (parts per billion) of cadmium metal ions in water at near-infrared wavelengths using established techniques for the detection of specific probe-target biomarker conjugation chemistries.

The stereoscopic image is often captured using dual cameras arranged side-by-side and optical path switching systems such as two separate solid lenses or biprism/mirrors. The miniaturization of the overall size of current stereoscopic devices down to several millimeters is at a sacrifice of further device size shrinkage. The limited light entry worsens the final image resolution and brightness. It is known that optofluidics offer good re-configurability for imaging systems. Leveraging this technique, we report a reconfigurable optofluidic system whose optical layout can be swapped between a singlet lens with 10 mm in diameter and a pair of binocular lenses with each lens of 3 mm in diameter for switchable two-dimensional (2D) and three-dimensional (3D) imaging. The singlet and the binoculars share the same optical path and the same imaging sensor. The singlet acquires a 3D image with better resolution and brightness, while the binoculars capture stereoscopic image pairs for 3D vision and depth perception. The focusing power tuning capability of the singlet and the binoculars enable image acquisition at varied object planes by adjusting the hydrostatic pressure across the lens membrane. The vari-focal singlet and binoculars thus work interchangeably and complementarily. The device is thus expected to have applications in robotic vision, stereoscopy, laparoendoscopy and miniaturized zoom lens system.

In comparison to more developed optical method for microparticle manipulation like optical tweezers, an optopiezoelectric actuating system could provide force output that is several orders higher. Taking advantages of photoconductive materials, the concept of integrating a virtual electrode in a distributed opto-piezoelectric actuators was developed for real-time in-situ spatial tailoring for vast varieties of applications in biochips, smart structures, etc. In this study, photoconductive material titanium oxide phthalocyanine (TiOPc) was used as the active ingredient to enable the virtual electrode in an opto-piezoelectric material based distributed actuator. By illuminating light of proper wavelength and enough intensity onto TiOPc photoconductive material, the effective impedance of the illuminated portion of TiOPc could drop significantly. The contributions of using additives in the TiOPc photoconductive electrode to adjust the electrical properties was investigated for optimization. Further, the two-mode excited linear ultrasonic motor was also studied and the feasibility to integrate the TiOPc photoconductive electrode was discussed. The flexibility provided by this newly developed system could potential deliver versatile performance in biochip applications.

Surface-enhanced Raman spectroscopy (SERS) has immerged as a power analytical and sensing technique for many applications in biomedical diagnosis, life sciences, food safety, and environment monitoring because of its molecular specificity and high sensitivity. The inactive Raman scattering of water molecule makes SERS a suitable tool for studying biological systems. Microfluidic devices have also attracted a tremendous interest for the aforementioned applications. By integrating SERS-active substrates with microfluidic devices, it offers a new capability for in situ investigation of biological systems, their dynamic behaviors, and response to drugs or microenvironment changes. In this work, we designed and fabricated a microfluidic device with SERS-active substrates surrounding by cell traps in microfluidic channels for in situ study of live cells using SERS. The SERS-active substrates are quasi-3D plasmonic nanostructure array (Q3D-PNA) made in h-PDMS/PMDS with physically separated gold film with nanoholes op top and gold nanodisks at the bottom of nanowells. 3D finite-difference time-domain (3D-FDTD) electromagnetic simulations were performed to design Q3D-PNAs with the strongest local electric fields (hot spots) at the top or bottom water/Au interfaces for sensitive analysis of cells and small components, respectively. The Q3D-PNAs with the hot spots on top and bottom were placed at the up and down stream of the microfluidic channel, respectively. Each Q3D-PNA pattern was surrounded with cell trapping structures. The microfluidic device was fabricated via soft lithography. We demonstrated that normal (COS-7) and cancer (HpeG2) cells were captured on the Q3D-PNAs and investigated in situ using SERS.

Procalcitonin (PCT) is an early and highly specific biomarker in response to bacterial infection. The PCT-guided antibiotic therapy has demonstrated to be more efficient than standard therapy to reduce in antibiotic use without adverse outcome in mortality. The PCT detection in clinics is required to be highly sensitive with a sensitivity of 0.5 ng/ml. At present, the technologies for PCT detection are limited. This paper reported a highly sensitive nanoimprinted gold nanopillar array chip for PCT detection. To achieve high sensitivity for PCT detection, the gold nanopillar array sensing chip was designed by plasmonic simulation and fabricated by high fidelity nanoimprinting technology. The gold nanopillars of 140 nm were nanoimprinted on glass substrate. A robust sandwich bioassay of capture antibody /PCT / quantum dot (QD) conjugated detection antibody was established on the gold nanopillar array chip to detect PCT. The nanopillars serve as localized surface plasmon resonance (LSPR) generators to enhance the fluorescent emission from QD. A limit of detection (LOD) of 0.5 ng/ml was achieved for PCT detection. This is the first time that PCT is detected with such high sensitivity by LSPR enhanced QD emission. By considering the low-cost, high sensitivity of the bioassay, as well as the inexpensive mass fabrication of the high quality chips, this novel nanoimprinted gold nanopillar array chip is particularly useful for developing a point-of-care system for PCT detection.

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.

In this project we present a microfluidic platform with in-channel micro-electrodes for in situ screening of bio/chemical samples through a lab-on-chip system. We used a novel method to incorporate electrochemical sensors array (16x20) connected to a PCB, which opens the way for imaging applications. A 200 μm height microfluidic channel was bonded to electrochemical sensors. The micro-channel contains 3 inlets used to introduce phosphate buffer saline (PBS), ferrocynide and neurotransmitters. The flow rate was controlled through automated micro-pumps. A multiplexer was used to scan electrodes and perform individual cyclic voltammograms by a custom potentiostat. The behavior of the system was linear in terms of variation of current versus concentration. It was used to detect the neurotransmitters serotonin, dopamine and glutamate.

We report a concept for the simple fabrication of easy-to-use chips for immunoassays in the context of point-of-care diagnostics. The chip concept comprises mainly three features: (1) the efficient integration of reagents using beads functionalized with receptors, (2) the generation of capillary-driven liquid flows without using external pumps, and (3) a high-sensitivity detection of analytes using fluorescence microscopy. We fabricated prototype chips using dry etching of Si wafers. 4.5-μm-diameter beads were integrated into hexagonal arrays by sedimentation and removing the excess using a stream of water. We studied the effect of different parameters and showed that array occupancies from 30% to 50% can be achieved by pipetting a 250 nL droplet of 1% bead solution and allowing the beads sediment for 3 min. Chips with integrated beads were sealed using a 50-μm-thick dry-film resist laminated at 45 °C. Liquids pipetted to loading pads were autonomously pulled by capillary pumps at a rate of 0.35 nL s^-1 for about 30 min. We studied ligand-receptor interactions and binding kinetics using time-lapse fluorescence microscopy and demonstrated a 5 pM limit of detection (LOD) for an anti-biotin immunoassay. As a clinically-relevant example, we implemented an immunoassay to detect prostate specific antigen (PSA) and showed an LOD of 108 fM (i.e. 3.6 pg mL^-1). While a specific implementation is provided here for the detection of PSA, we believe that combining capillary-driven microfluidics with arrays of single beads and fluorescence readout to be very flexible and sufficiently sensitive for the detection of other clinically-relevant analytes.

Microfluidics has emerged in recent years as a versatile method of manipulating fluids at small length-scales, and in particular, for generating and manipulating micron size droplets with controllable size and functionality. For example, many research groups developed microfluidics devices for cell encapsulation, and synthesizing functionalized polymer microspheres and inorganic nanoparticles with precise control over their shapes and sizes. In this talk, I will showcase 2 microfluidic platforms to highlight their versatility and potential biomedical applications. (1) Droplet microfluidic platforms (a) A droplet microfluidics method to fabricate alginate microspheres while simultaneously immobilizing anti-Mycobacterium tuberculosis complex IgY and anti-Escherichia coli IgG antibodies primarily on the porous alginate carriers for specific binding and binding affinity tests. The binding affinity of antibodies is directly measured by fluorescence intensity of stained target bacteria on the microspheres. We demonstrate that the functionalized alginate microspheres yield specificity comparable with an enzyme-linked immunosorbent assay. We can easily modify the size and shape of alginate microspheres, and increase the concentration of functionalized alginate microspheres to further enhance binding kinetics and enable multiplexing. (b) A novel droplet microfluidics method to image oxygen in single islets (pancreatic cells) for glucose sensing. Individual islets and a fluorescent oxygen-sensitive dye were encased within a thin alginate polymer microcapsule for insulin secretion monitoring. The sensing system operated similarly from 2-48 hours following encapsulation, and viability and function of the islets were not significantly affected by the encapsulation process. This approach should be applicable to other cell types and dyes sensitive to other biologically important molecules. (2) A microfluidic chamber to perform uniform electric field stimulation in circular shaped culturewares A 3D computer-aided designed (CAD) polymeric insert is designed and retrofitted to circular shaped culturewares in an integrated microfluidic electrical stimulation platform to generate uniform EF with higher cell yields. In particular, NIH/3T3 mouse embryonic fibroblast cells are used to validate the performance of the 3D designed Poly(methyl methacrylate) (PMMA) inserts in a circular-shaped 6-well plate. The CAD based inserts can be easily scaled up to further increase effective stimulation area percentages, and also be implemented in commercially available culturewares for a wide variety of EF-related research such as EF-cell interaction and tissue regeneration studies.

Signal transductions including multiple protein post-translational modifications (PTM), protein-protein interactions (PPI), and protein-nucleic acid interaction (PNI) play critical roles for cell proliferation and differentiation that are directly related to the cancer biology. Traditional methods, like mass spectrometry, immunoprecipitation, fluorescence resonance energy transfer, and fluorescence correlation spectroscopy require a large amount of sample and long processing time. “microchannel for multiple-parameter analysis of proteins in single-complex (mMAPS)”we proposed can reduce the process time and sample volume because this system is composed by microfluidic channels, fluorescence microscopy, and computerized data analysis. In this paper, we will present an automated mMAPS including integrated microfluidic device, automated stage and electrical relay for high-throughput clinical screening. Based on this result, we estimated that this automated detection system will be able to screen approximately 150 patient samples in a 24-hour period, providing a practical application to analyze tissue samples in a clinical setting.

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.

In wounds, cells secret biomolecules such as vascular endothelial growth factor (VEGF), a protein that controls many processes in healing. VEGF protein is expressed in a gradient in tissue, and its shape will be affected by the tissue injury sustained during wounding. In order to study the responses of keratinocyte cell migration to VEGF gradients and the geometric factors on wound healing, we designed a microfluidic gradient device that can generate large area gradients (1.5 cm in diameter) capable of mimicking arbitrary wound shapes. Microfluidic devices offer novel techniques to address biological and biomedical issues. Different from other gradient microfluidics, our device balances diffusion of biomolecules versus the convective clearance by a buffer flow on the opposite ends of the gradient. This allows us to create a large area gradient within shorter time scales by actively driving mass transport. In addition, the microfluidic device makes use of a porous filter membrane to create this balance as well as to deliver the resulting gradient to a culture of cells. The culture of cells are seeded above the gradient in a gasket chamber. However, Keratinocytes do not migrate effectively on filter paper. Therefore, in order to improve the motility of cells on the surface, we coated the filter paper with a 30m thick layer of gelatin type B. after observation under the microscope we found that the gelatin coated sample showed cells with more spread out morphology, with 97% viability, suggesting better adhesion than the non-coated sample.

The miniaturization, rapid prototyping and automation of lab-on-a-chip technology play nowadays a very important role. Lab-on-a-chip technology is successfully implemented not only for environmental analysis and medical diagnostics, but also as replacement of animals used for the testing of substances in the pharmaceutical and cosmetics industries. For that purpose the Fraunhofer IWS and partners developed a lab-on-a-chip platform for perfused cell-based assays in the last years, which includes different micropumps, valves, channels, reservoirs and customized cell culture modules. This technology is already implemented for the characterization of different human cell cultures and organoids, like skin, liver, endothelium, hair follicle and nephron. The advanced universal lab-on-a-chip platform for complex, perfused 3D cell cultures is divided into a multilayer basic chip with integrated micropump and application-specific 3D printed cell culture modules. Moreover a technology for surface modification of the printed cell culture modules by laser micro structuring and a complex and flexibly programmable controlling device based on an embedded Linux system was developed. A universal lab-on-a-chip platform with an optional oxygenator and a cell culture module for cubic scaffolds as well as first cell culture experiments within the cell culture device will be presented. The module is designed for direct interaction with robotic dispenser systems. This offers the opportunity to combine direct organ printing of cells and scaffolds with the microfluidic cell culture module. The characterization of the developed system was done by means of Micro-Particle Image Velocimetry (μPIV) and an optical oxygen measuring system.

The NANIVID – or Nano Intravital Device – is an implantable delivery tool designed to locally affect the tumor microenvironment in vivo. This technology is being redesigned and validated as a cell collection tool for the study of metastatic cancer cells. A methodology has been developed to facilitate this transition, consisting of microfluidic analysis of the device microchannels and a series of cell-related collection experiments building up to in vivo collection. Single-chamber designs were first used to qualitatively demonstrate the feasibility of cell collection ex vivo. This was followed by the development and implementation of devices containing a second, negative-control chamber for quantitative analysis. This work sets the foundation for in vivo cancer cell migration studies utilizing the NANIVID.