This PDF file contains the front matter associated with SPIE Proceedings Volume 8976, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

Biophotonic and Life Science applications often require non-CMOS compatible materials to be patterned with sub μm resolution. Whilst the mass production of sub μm patterns is well established in the semiconductor industry, semiconductor fabs are limited to using CMOS compatible materials. IMT of Switzerland has implemented a fully automated manufacturing line that allows cost effective mass manufacturing of consumables for biophotonics in substrate materials like D263 glass or fused silica and layer/coating materials like Cr, SiO₂, Cr₂O₅, Nb₂O₅, Ta₂O₅ and with some restrictions even gold with sub-μm patterns. The applied processes (lift-off and RIE) offer a high degree of freedom in the design of the consumable.

One potential problem with microfluidic systems is the accumulation of particles and fluid bubbles inside chambers and other structures, which causes distortion in fluid flow potentially leading to device or system failure. Microfluidic channels and chambers that utilize a “cathedral-ceiling” arrangement, whereby periodic posts support the tops of the channels, have been suggested to improve defect tolerance over arrays of parallel channels through the provision of multiple paths during localized blockage formation. This paper builds on our prior investigations through development of a combined rule-based defect placement system and Monte Carlo method for modeling the fluid dynamics and blockage formation based on the likelihood of blockages forming in areas of high particle traffic and low flow. Our COMSOL model generates 150 randomly (normal) distributed particle streamlines. Coordinates along these streamlines are crossexamined to find the lowest flow areas, which are deemed likely points for blockage formation. MATLAB filtering then determines which microfluidic channel areas are most likely to obstruct based on particle population density. This process is iterated as blockages form, creating new streamline patterns, which in turn indicate placements for new blockages, and modified geometry for successive modelling iterations. This semi-automated method has enabled us to predict where the particles may accumulate and how this progressive block formation may change system pressure and flow. Results obtained support the findings of significantly increased lifetime expectancy of microfluidic chambers with periodic posts compared to arrays of parallel channels, while also providing greater insight into where blockages may form in the cathedral-ceiling type geometry.

OSTE polymer has the aim to address today’s dissemination gap between successful lab-on-chip research and the healthcare setting. We have formulated and demonstrated a novel, superior, polymer system, OSTE, and its manufacturing platform, which is based on the mixture of three monomers: thiols, -enes and epoxies. The uniqueness of the OSTE approach stems from the curing in two distinct steps: after the first cure, an intermediate polymer is formed which is ideally suited for surface modifications and bonding; after the second cure we obtain an inert and robust polymer. Our vision is that OSTE has the potential to form a de-facto standard for research and development of high performance labs-on-chip in academia and industry.

In this paper we present a fast, low cost bonding technology for combining rigid epoxy components with soft membranes made out of polydimethylsiloxane (PDMS). Both materials are commonly used for microfluidic prototyping. Epoxy resins are often applied when rigid channels are required, that will not deform if exposed to high pressure. PDMS, on the other hand, is a flexible material, which allows integration of membrane valves on the chip. However, the integration of pressure driven components, such as membrane valves and pumps, into a completely flexible device leads to pressure losses. In order to build up pressure driven components with maximum energy efficiency a combination of rigid guiding channels and flexible membranes would be advisable. Stereolithographic (STL) structuring would be an ideal fabrication technique for this purpose, because complex 3D-channels structures can easily be fabricated using this technology. Unfortunately, the STL epoxies cannot be bonded using common bonding techniques. For this reason we propose two UV-light based silanization techniques that enable plasma induced bonding of epoxy components. The entire process including silanization and corona discharge bonding can be carried out within half an hour. Average bond strengths up to 350 kPa (depending on the silane) were determined in ISO-conform tensile testing. The applicability of both techniques for microfluidic applications was proven by hydrolytic stability testing lasting more than 40 hours.

Microfluidic devices offer novel techniques to address biological and biomedical issues. Standard microfluidic fabrication uses photolithography to pattern channels on silicon wafers with high resolution. Even the relatively straightforward SU8 and soft lithography in microfluidics require investing and training in photolithography, which is also time consuming due to complicated thick resist procedures, including sensitive substrate pretreatment, coating, soft bake, expose, post-exposure bake, and developing steps. However, for applications where low resolution (>200 μm) and high turn-around (> 4 designs/day) prototyping are met with little or no lithography infrastructure, robotic cutters [1] offer flexible options for making glass and PDMS microfluidics. We describe the use of robotics cutters for designing microfluidic geometries, and compliment it with safe glass etching, with depths down to 60 μm. Soft lithography patterning of 200 μm thick PDMS membrane was also explored. Without high equipment investment and lengthy student training, both glass and PDMS microfluidics can be achieved in small facilities using this technique.

Microfluidics provides exciting possibilities for miniaturized biosensors systems allowing for highly parallel automated high throughput tests to be performed. Detection of low concentrations of bacteria, viral particles and parasites in food samples is a challenging process. The capture of the target can be more effectively carried out with efficient mixing. We present a simple microfluidic system capable of controlled transport of rotating magnetic beads among soft magnetic patterns. Low aspect ratio NiFe discs (200 nm tall, diameter 3 μm) are patterned onto a silicon wafer. A PDMS channel is bonded onto the wafer to create the microfluidic channel. An external permanent magnet attached to a motor provides a magnetic field, which can be rotated at different speeds while magnetizing the NiFe disks in the channel. Microbeads (Dynabeads M-280, Invitrogen) introduced into the channel with a syringe pump are trapped at the poles of the now magnetized soft magnetic discs. Rotation of the external permanent magnet induced magnetic poles in the soft magnetic discs which will in turn rotate the trapped microbeads. We have already demonstrated the capacity to capture particles from flow with rotating M-280 beads in this device.

This paper presents a new fabrication technique to achieve ultra high-aspect ratio artificial cilia micro-patterned from flexible highly magnetic rare earth nanoparticle-doped polymers. We have developed a simple, inexpensive and scalable fabrication method to create cilia structures that can be actuated by miniature electromagnets, that are suitable to be used for lab-on-a chip (LOC) and micro-total-analysis-system (μ-TAS) applications such as mixers and flow-control elements. The magnetic cilia are fabricated and magnetically polarized directly in microfluidic channels or reaction chambers, allowing for easy integration with complex microfluidic systems. These cilia structures can be combined on a single chip with other microfluidic components employing the same permanently magnetic nano-composite polymer (MNCP), such as valves or pumps. Rare earth permanent magnetic powder, (Nd_0.7Ce_0.3)_10.5Fe_83.9B_5.6, is used to dope polydimethylsiloxane (PDMS), resulting in a highly flexible M-NCP of much higher magnetization and remanence [1] than ferromagnetic polymers typically employed in magnetic microfluidics. Sacrificial poly(ethylene-glycol) (PEG) is used to mold the highly magnetic polymer into ultra high-aspect ratio artificial cilia. Cilia structures with aspect ratio exceeding 8:0.13 can be easily fabricated using this technique and are actuated using miniature electromagnets to achieve a high range of motion/vibration.

Within the last decades more and more microfluidic systems for applications in chemistry, biology or medicine were developed. Most of them need a connection between the chip and its macroscopic environment e.g., pumps. Numerous concepts for such interconnections are known from literature but most of them allow only a small number of connections and are neither chemically inert nor contamination-free. We developed a chemically inert, reusable, multichannel Chipto- World-Interface (CWI) based on a force fit connection. This principle is comparable to hollow screws as used in highperformance liquid chromatography. The CWI can be used to connect chips, made of different materials, e.g., glass, polydimethylsiloxane (PDMS), or epoxy polymers, with up to 100 thermoplastic tubes. The dimensions of the CWI and the number of connections can be individually adapted depending on the chip dimensions but the pitch between the tubes is fixed. Due to the design of the CWI the fluid is only in contact with the chip and the tubing material, thus leading to a contamination free and zero dead volume interconnection. Using tubes of polytetrafluorethylene (PTFE, Teflon®) even enables probing with organic solvents like dimethylformamide, dichloromethane or tetrahydrofuran over several hours without leakage or corrosion of the CWI. During experiments the CWI with 100 connections resisted pressure up to 630 kPa (6.3 bar) and sustained flow rates higher than 4 ml/min.

We demonstrate the feasibility of trapping and manipulating individual macromolecules such as globular proteins (~ 5 nm in diameter) in free solution using a flow-based, microfluidic confinement method. This new method enables confinement of small nanoscale objects in free solution by utilizing a planar extensional flow created at a microchannel junction. The fluid flow based confinement method described in this work expands the micro/nanomanipulation toolbox by offering a powerful and versatile platform for non-perturbative observation and analysis of single macromolecules in free solution without force fields (electrical, magnetic, optical and acoustic) and surface immobilization.

Since the early days microfluidics as a scientific discipline has been an interdisciplinary research field with a wide scope of potential applications. Besides tailored assays for point-of-care (PoC) diagnostics, microfluidics has been an important tool for large-scale screening of reagents and building blocks in organic chemistry, pharmaceutics and medical engineering. Furthermore, numerous potential marketable products have been described over the years. However, especially in industrial applications, microfluidics is often considered only an alternative technology for fluid handling, a field which is industrially mostly dominated by large-scale numerically controlled fluid and liquid handling stations. Numerous noteworthy products have dominated this field in the last decade and have been inhibited the widespread application of microfluidics technology. However, automated liquid handling stations and microfluidics do not have to be considered as mutually exclusive approached. We have recently introduced a hybrid fluidic platform combining an industrially established liquid handling station and a generic microfluidic interfacing module that allows probing a microfluidic system (such as an essay or a synthesis array) using the instrumentation provided by the liquid handling station. We term this technology “Microfluidic on Liquid Handling Stations (μF-on-LHS)” – a classical “best of both worlds”- approach that allows combining the highly evolved, automated and industry-proven LHS systems with any type of microfluidic assay. In this paper we show, to the best of our knowledge, the first droplet microfluidics application on an industrial LHS using the μF-on-LHS concept.

There is currently no global screening system to detect low quality pharmaceuticals, despite widespread recognition of the public health problems caused by substandard and falsified medicines. In order to fill this void, we designed a rapid field screening test that is interfaced with the mobile phone network. The user scrapes a pill over several reaction areas on a paper test card, and then dips one edge of the card into water to activate dried reagents stored on the paper. These reagents carry out multiple color tests and result in a pattern of colored stripes that give information about the chemical content of the pill. The test cards are inexpensive and instrument-free, and we think they will be a scalable testing option in low resource settings. Studies on falsified drugs archived at the FDA show that the test cards are effective at detecting a wide variety of low-quality formulations of many classes of pharmaceuticals, and field tests are currently under way in Kenya.

Electrical manipulation of biological samples using glass-based electrofluidics fabricated by femtosecond laser, in which the microfluidic structures are integrated with microelectric components, is presented. Electro-orientation of movement of living cells with asymmetric shapes such as Euglena gracilis of aquatic microorganisms in microfluidic channels is demonstrated using the fabricated electrofluidics. By integrating the properly designed microelectrodes into microfluidic channels, the orientation direction of Euglena cells can be well controlled.

Cultures of algae, such as cyanobacteria, are a promising source of renewable energy. However, algal growth is highly dependent on light intensity and standard photobioreactors do a poor job of distributing light uniformly for algal utilization due to shading effects in dense algal cultures. Engineered scattering schemes are already employed in current slab-waveguide technologies, like edge-lit LEDs. Stacking such slab-waveguides that uniformly distribute light could potentially yield photobioreactors to overcome the shading effect and grow extremely high densities of algal cultures that would lower monetary and energetic costs. Here, we characterize and design a scattering scheme for specific application within photobioreactors which employs a gradient distribution of surface scatterers with uniform lateral scattering intensity. This uniform scattering scheme is shown to be superior for algal cultivation.

Manipulation, sorting and recovering of specific live cells from samples containing less than a few thousand cells is becoming a major hurdle in rare cell exploration such as stem cell research or cell based diagnostics. Moreover the possibility of recovering single specific cells for culturing and further analysis would be of great impact in many biological fields ranging from regenerative medicine to cancer therapy. In recent years considerable effort has been devoted to the development of integrated and low-cost optofluidic devices able to handle single cells, which usually rely on microfluidic circuits that guarantee a controlled flow of the cells. Among the different microfabrication technologies, femtosecond laser micromachining (FLM) is ideally suited for this purpose as it provides the integration of both microfluidic and optical functions on the same glass chip leading to monolithic, robust and portable devices. Here a new optofluidic device is presented, which is capable of sorting and recovering of single cells, through optical forces, on the basis of their fluorescence and. Both fluorescence detection and single cell sorting functions are integrated in the microfluidic chip by FLM. The device, which is specifically designed to operate with a limited amount of cells but with a very high selectivity, is fabricated by a two-step process that includes femtosecond laser irradiation followed by chemical etching. The capability of the device to act as a micro fluorescence-activated cell sorter has been tested on polystyrene beads and on tumor cells and the results on the single live cell recovery are reported.

For many problems in system biology or pharmacology, in-vivo-like models of cell-cell interactions or organ functions are highly sought after. Conventional stationary cell culture in 2D plates quickly reaches its limitations with respect to an in-vivo like expression and function of individual cell types. Microfabrication technologies and microfluidics offer an attractive solution to these problems. The ability to generate flow as well as geometrical conditions for cell culture and manipulation close to the in-vivo situation allows for an improved design of experiments and the modeling of organ-like functionalities. Furthermore, reduced internal volumes lead to a reduction in reagent volumes necessary as well as an increased assay sensitivity. In this paper we present a range of microfluidic devices designed for the co-culturing of a variety of cells. The influence of substrate materials and surface chemistry on the cell morphology and viability for long-term cell culture has been investigated as well as strategies and medium supply for on-chip cell cultivation.

In this study, a surface acoustic wave (SAW)-based microfluidic device has been developed to separate heterogeneous particle or cell mixtures in a continuous flow using acoustophoresis. The microfluidic device is comprised of two components, a SAW transducer and a microfluidic channel made of polydimethylsiloxane (PDMS). The SAW transducer was fabricated by patterning two pairs of interdigital electrodes on a lithium niobate (LiNbO₃) piezoelectric substrate. When exciting the SAW transducer by AC signals, a standing SAW is generated along the cross-section of the channel. Solid particles immersed in the standing SAW field are accordingly pushed to the pressure node arising from the acoustic radiation force acting on the particles, referring to the acoustic particle-focusing phenomenon. Acoustic radiation force highly depends on the particle properties, resulting in different acoustic responses for different types of cells. A numerical model, coupling the piezoelectric effect in the solid substrate and acoustic pressure in the fluid, was developed to provide a better understanding of SAW-based particle manipulation. Separation of two types of fluorescent particles has been demonstrated using the developed SAW-based microfluidic device. An efficient separation of E. coli bacteria from peripheral blood mononuclear cell (PBMC) samples has also been successfully achieved. The purity of separated E. coli bacteria and separated PBMCs were over 95% and 91%, respectively, obtained by a flow cytometric analysis. The developed microfluidic device can efficiently separate E. coli bacteria from biological samples, which has potential applications in biomedical analysis and clinical diagnosis.

The design of a novel therapeutic drug monitoring (TDM) point-of-care-testing (POCT) biochip for immunosuppressants detection in transplanted patients is described. The chip consists of two polymeric parts, a top PMMA slide and a bottom ZEONOR® thin foil, bonded together by means of a pressure sensitive adhesive tape. The tape, with lower refractive index, is shaped in order to obtain a microfluidic multi-channel array. The optical signal, coming from an external light source and travelling along the ZEONOR® thin foil, excites the fluorescent sensing layer immobilized onto the fluidic channels. Preliminary tests with the bioassay implementation for tacrolimus detection are reported.

Microencapsulation of multiple drugs and imaging agents is significant for various biomedical applications. In this work we describe a novel method based on flow focusing geometry using tri-axial metallic capillary needles manufactured by a laser beam welding process. The tri-axial needle can be readily cleaned, assembled, and aligned. With this needle assembly, we develop a tri-axial capillary flow focusing device in which different combinations of liquids are focused in the core of a high-speed gas stream coflowing through a small orifice. Under appropriate working conditions, stable cone-jet configurations with three layers of liquids in an external gas stream can be obtained, resulting in multilayered microparticles with outer shell, middle layer, and inner core. The new design of tri-axial needles enables reliable encapsulation of multiple drugs and imaging agents in biodegradable microcapsules with the enhanced size distribution, increased productivity, and improved drug-loading efficiency. Furthermore, in this method the outer and the middle shell fluids can be released to produce monodisperse microparticles at smaller scales which have potential applications in multi-modal imaging, drug delivery, material processing and biomedicine.

A minimally-invasive diagnosis of pancreatic cancer is accomplished by obtaining a fine needle aspirate and observing the cell preparations under conventional optical microscopy. As an unavoidable artifact, native tissue architecture is lost, making definite diagnosis of malignancy, or invasive neoplasm, impossible. One solution is the preparation of core biopsies (CBs) within a microfluidic device that are subsequently imaged in 3D. In this paper, porcine pancreas CBs (L = 1-2 cm, D = 0.4-2.0 mm) were formalin-fixed, stained and optically cleared (FocusClear®). In brightfield at 40x, light transmission through the ordinarily opaque CBs was increased 5-15x, and internal islet structures were easily identified 250-300 μm beneath the tissue surface. Typically, specimen preparation is time intensive and requires precise handling since CBs are delicate; thus, fixative, absorptive stain and FocusClear® diffusion were done slowly and manually. To significantly speed up tissue processing, we developed a microfluidic device consisting of both a main channel (L = 12.5 cm, D = 1.415 mm) with a circular cross section used for fixing and transporting the CB and an intersecting U-channel employed for staining. Space between the CB and channel wall provided a key feature not traditionally employed in microfluidic devices, such that at low flow rates (5-10 mL/min) CBs were fixed and stained while the specimen remained stationary. By switching quickly to higher flow rates (15-20 mL/min), we could precisely overcome adhesion and transport the specimen within the channel towards the imaging platform for 3D pathology.

Passive flow regulators are usually intended to deliver or drain a fluid at a constant rate independently from pressure variations. New designs of passive flow regulators made of a stack of a silicon membrane anodically bonded to a Pyrex substrate are proposed. A first design has been built for the derivation of cerebrospinal fluid (CSF) towards peritoneum for hydrocephalus treatment. The device allows draining CSF at the patient production rate independently from postural changes. The flow rate is regulated at 20 ml/h in the range 10 to 40 mbar. Specific features to adjust in vivo the nominal flow rate are shown. A second design including high pressure shut-off feature has been made. The intended use is drug delivery with pressurized reservoir of typically 100 to 300 mbar. In both cases, the membrane comprises several holes facing pillars in the Pyrex substrate. These pillars are machined in a cavity which ensures a gap between the membrane and the pillars at rest. The fluid in the pressurized reservoir is directly in contact with the top surface of the membrane, inducing its deflection towards Pyrex substrate and closing progressively the fluidic pathway through each hole of the membrane. Since the membrane deflection is highly non-linear, FEM simulations have been performed to determine both radial position and diameter of the membrane holes that ensure a constant flow rate for a given range of pressure.

Optical forces are used to fixate biological cells with optical tweezers where numerous biological parameters and phenomena can be studied. Optical beams carry a small momentum which generates a weak optical force, but on a cellular level this force is strong enough to allow for manipulation of biological cells in microfluidic systems exclusively using light. We demonstrate an optical cell sorter that uses simultaneous manipulation by multiple laser beams using the Generalized Phase Contrast method (GPC). The basic principle in an optical sorter is that the radiation force of the optical beam can push the biological cell from one microfluidic sheath flow to another. By incorporating a spatial light modulator the manipulation can be made parallel with multiple laser beams. We claim advantages over the serial optical sorters with only a single laser beam that has been demonstrated by others.

Since their invention by Ashkin optical tweezers have demonstrated their ability and versatility as a non-invasive tool for micromanipulation. One of the most useful additions to the basic optical tweezers system is micro-Raman spectroscopy, which permits highly sensitive analysis of single cells or particles. We report on the development of a dual laser system combining two spatial light modulators to holographically manipulate multiple traps (at 1064nm) whilst undertaking Raman spectroscopy using a 532nm laser. We can thus simultaneously trap multiple particles and record their Raman spectra, without perturbing the trapping system. The dual beam system is built around micro-fluidic channels where crystallisation of calcium carbonate occurs on polymethylmethacrylate (PMMA) beads. The setup is designed to simulate at a microscopic level the reactions that occur on items in a dishwasher, where permanent filming of calcium carbonate on drinking glasses is a problem. Our system allows us to monitor crystal growth on trapped particles in which the Raman spectrum and changes in movement of the bead are recorded. Due to the expected low level of crystallisation on the bead surfaces this allows us to obtain results quickly and with high sensitivity. The long term goal is to study the development of filming on samples in-situ with the microfl.uidic system acting as a model dishwasher.

The existing technological approaches employed in the realization of optical sensors still face two major challenges: the inherent inability of most sensors to integrate the optical source in the transducer chip, and the need to specifically design the optical transducer per application. We have introduced a unique Optoelectronic chip that consists of a series of light emitting diodes (LEDs) coupled to silicon nitride waveguides allowing for multi-analyte detection. Each optocoupler is structured as Broad-Band Mach-Zehnder Interferometer and has its own excitation source and can either have its own detector or the entire array can share a common detector. The light emitting devices (LEDs) are silicon avalanche diodes which when biased beyond their breakdown voltage emit in the VIS-NIR part of the spectrum. The optoelectronic chip is fabricated by standard silicon technology allowing for potential mass production in silicon foundries. The integrated nature of the optoelectronic chip and the ability to functionalize each transducer independently allows for the development of miniaturized optical transducers tailored towards multi-analyte tests. The platform has been successfully applied in bioassays and binding assays monitoring in a real-time and label-free format and is currently being applied to ultra-sensitive food safety applications.

This work reports our efforts on developing simple-to-use microfluidic devices for point-of-care diagnostic applications with recent extensions that include the trapping of microbeads using dielectrophoresis (DEP) and the modulation of capillary-driven flow using integrated microheaters. DEP serves the purpose of trapping microbeads coated with receptors and analytes for detection of a fluorescent signal. The microheater is actuated once the chip is filled by capillarity, creating an evaporation-induced flow tuned according to assay conditions. The chips are composed of a glass substrate patterned with 50-nm-thick Pd electrodes and microfluidic structures made using a 20-μm-thick dry-film resist (DFR). Chips are covered/sealed by low-temperature (50 °C) lamination of a 50-μm-thick DFR layer having excellent optical and mechanical properties. To separate cleaned and sealed chips from the wafer, we used an effective chip singulation technique that we informally call the "chip-olate" process. In the experimental section, we first studied dielectrophoretic trapping of 10 μm beads for flow rates ranging from 80 pL s^-1 to 2.5 nL s^-1 and that are generated by an external syringe pump. Then, we characterized the embedded microheater in DFR-covered chips. Flow rates as high as 8 nL s^-1 were generated by evaporation-induced flow when the heater was biased by 10 V, corresponding to 270 mW power. Finally, DEP-based trapping and fluorescent detection of functionalized beads were demonstrated as the flow was generated by the combination of capillary filling and evaporation-induced flow.

A central method in a standard biochemical laboratory is represented by the polymerase chain reaction (PCR), therefore many attempts have been performed so far to implement this technique in lab-on-a-chip (LOC) devices. PCR is an ideal candidate for miniaturization because of a reduction of assay time and decreased costs for expensive bio-chemicals. In case of the “classical” PCR, detection is done by identification of DNA fragments electrophoretically separated in agarose gels. This method is meanwhile frequently replaced by the so-called Real-Time-PCR because here the exponential increase of amplificates can be observed directly by measurement of DNA interacting fluorescent dyes. Two main methods for on-chip PCRs are available: traditional “batch” PCR in chambers on a chip using thermal cycling, requiring about 30 minutes for a typical PCR protocol and continuous-flow PCR, where the liquid is guided over stationary temperature zones. In the latter case, the PCR protocol can be as fast as 5 minutes. In the presented work, a proof of concept is demonstrated for a real-time-detection of PCR products in microfluidic systems.

We introduce a novel and cost-effective capillary gel electrophoresis (CGE) system utilizing disposable pen-shaped gelcartridges for highly efficient, high speed, high throughput fluorescence detection of bio-molecules. The CGE system has been integrated with dual excitation and emission optical-fibers with micro-ball end design for fluorescence detection of bio-molecules separated and detected in a disposable pen-shaped capillary gel electrophoresis cartridge. The high-performance capillary gel electrophoresis (CGE) analyzer has been optimized for glycoprotein analysis type applications. Using commercially available labeling agent such as ANTS (8-aminonapthalene-1,3,6- trisulfonate) as an indicator, the capillary gel electrophoresis-based glycan analyzer provides high detection sensitivity and high resolving power in 2-5 minutes of separations. The system can hold total of 96 samples, which can be automatically analyzed within 4-5 hours. This affordable fiber optic based fluorescence detection system provides fast run times (4 minutes vs. 20 minutes with other CE systems), provides improved peak resolution, good linear dynamic range and reproducible migration times, that can be used in laboratories for high speed glycan (N-glycan) profiling applications. The CGE-based glycan analyzer will significantly increase the pace at which glycoprotein research is performed in the labs, saving hours of preparation time and assuring accurate, consistent and economical results.

Tuberculosis is one of the most deadly diseases that kills over one million people each year and infects one-third of the world’s population. The disease is spread by infection with Mycobacterium tuberculosis (Mtb). Owing to its airborne transmission, early diagnosis is critical to the prevention and control of TB. Standard diagnostic methods, acid-fast smear from sputum, often do not become positive until after transmission occurs, which allows the spread of the disease. Culture-based techniques are more sensitive, but take weeks to obtain results because of the extremely slow growth rate of Mtb. In this study a new method to detect indicator enzyme based on the isolation of tubercle bacillus in a large number of picoliter droplets combined with a fluorescent probe has been developed. We use BlaC (an enzyme naturally expressed/secreted by tubercle bacilli) as a marker and a designed BlaC-specific fluorogenic substrates as probes for Mtb detection. We present here a new method to detect the indicator enzyme based on the isolation, digitization and concentration of bacteria samples in a large number of picoliter drops. We show that by controlling the size of the droplets we can control the rate of conversion. Hence rapid increase in signal has been observed as the size of the drops has been decreased. Our vision is that this tool will be able to detect tubercle bacilli in a sensitive, rapid, specific and quantitative manner in vitro at a low cost, particularly in resource limited settings where TB is the most prevalent.

An optofluidic jet waveguide for Raman spectroscopy is reported. In this device a micro-channel is used to produce a high speed liquid stream acting at the same time, as the solution to analyse and as an optical waveguide. The liquid waveguide, exploiting total internal reflection, is able to effectively collect the Raman signal produced by the chemical compound under analysis opportunely excited by means of a laser source. Using a self-aligned configuration, the liquid jet is directly coupled with a multimode optical fiber collecting the Raman signal towards the detection system. The waveguiding nature of a liquid jet enables high Raman signal collection and the device configuration allows strong reduction of the background as no confining walls are used to contain the solution to analyse. The performances of the system have been successfully tested on isopropyl alcohol in water solutions showing a detection limit for this chemical compound of 0.8±0.1%.

Chemical reactions are often described as a progression along a reaction coordinate. Waveguide evanescent fields generate an electromagnetic force that spans tens of nanometers and have been used previously to trap protein molecules. Applying this force along a reaction coordinate could radically alter the chemical reaction by modifying the activation energy or biasing the reaction towards a specific pathway. Here, we show that the adsorption of proteins onto carbon nanotubes can be controlled with opto-mechanical forces. An analytic model for the reaction was developed, the predictions of which were explored by probing the energy barrier under various experimental conditions.

A robust capacitive micromachined ultrasonic transducer has been developed. In this novel configuration, a stack of two deflectable membranes are suspended over a fixed bottom electrode. Similar to conventional capacitive ultrasonic transducers, a generated electrostatic force between the electrodes causes the membranes to deflect and vibrate. However, in this new configuration the transducer effective cavity height is reduced due to the deflection of two membranes. Therefore, the transducer spring constant is more susceptible to bias voltage, which in return reduces the required bias voltage. The transducers have been produced employing a MEMS sacrificial technique where two different membrane anchoring (curved- and flat- anchors) structures, with similar membrane radii were fabricated. Highly doped polysilicon was used as the membrane material. The resonant frequencies of the two transducers have been investigated. It was found that the transducers with curved membrane anchors exhibits a larger resonant frequency shift compared to the transducers with flat membranes for a given bias voltage. Comparison has been made between the spring constant of the flat membrane transducer and that of a conventional single membrane transducer. It is shown that the multiple moving membrane transducer exhibits a larger reduction in the spring constant compared to the conventional transducer, when driven with the same bias voltage. This results in a transducer with a higher power generation capability and sensitivity.

The newly developed configuration included adopting the photosensitive electrode material TiOPc (titanyl phthalocyanine) to create electrowetting on dielectric (EWOD) mechanism. With this new development, the electric potential on the surface of TiOPc could be on-line real-time changed and defined spatially by illuminating spatially distributed light beam patterns. We tried to control the polarized droplets in our EWOD devices by using different light intensities. The experimental results clearly demonstrated that the relationship of light intensity and electrowetting phenomena can provide us with a feasible platform to construct optofluidic chip with potential autonomous manipulation of samples for point-of-care home medical detection applications.

Label-free optical sensors are considered ideal for biomedical analysis since they provide the advantages of multiplex and real-time detection. They still suffer; however, from lower sensitivity and/or more sophisticated equipment as compared to indirect detection methods. Here, we propose a label-free sensor based on White Light Reflectance Spectroscopy that overcomes the limitation of high cost and low detection sensitivity. The optical setup consists of a VIS-NIR light source, a spectrometer and a reflection probe. The sensor is Si with 1-μm thick thermal SiO₂ and functionalized with antibodies. The incident light is directed vertically to sensor surface and the reflected interference spectrum is recorded through the spectrometer. The evolution of the biomolecular reactions are monitored in real-time by monitoring the shifts in the interference spectrum. Up to seven different reactions sites have been created onto the same sensing surface allowing for multi-analyte determinations. The analytical capabilities of the proposed sensor were demonstrated through the development of a sensitive immunoassay for the detection of C-Reactive Protein (CRP) in human serum samples. CRP, a biomarker related to acute inflammatory incidents, it has attracted particular interest as a marker of inflammation associated with cardiovascular diseases. The lowest CRP concentration detected was 10 ng/mL, and the dynamic range of the assay was extended up to 500 ng/mL. Regeneration of antibody coated sensing areas for up to 20 times without loss of immobilized antibody reactivity is also presented. In conclusion, the proposed sensing system is characterized by low cost, high assay sensitivity and, reliability.

In this manuscript we describe an electronic label-free method for detection of target cells, which has potential applications ranging from pathogen detection for food safety all the way to detection of circulating tumor cells for cancer diagnosis. The nanoelectronic platform consists of a stack of electrodes separated by a 30nm thick insulating layer. Cells binding to the tip of the sensor result in a decrease in the impedance at the sensing tip due to an increase in the fringing capacitance between the electrodes. As a proof of concept we demonstrate the ability to detect Saccharomyces Cerevisae cells with high specificity using a sensor functionalized with Concanavalin A. Ultimately we envision using this sensor in conjunction with a technology for pre-concentration of target cells to develop a fully integrated micro total analysis system.

Development of point-of-care biosensors continues to gain popularity due to the demand of improving the cost performance in today’s health care. As cardiovascular disease induced death remains on the top 3 death causes for most Asian countries, this paper is to present a high-sensitivity point-of-care biosensor for the detection of cardiovascular disease biomarkers. To meet the point-of-care biosensors requirements, which include characteristics such as small size, low cost, and ease of operation, we adopted electrochemical methods as the basis of detection. The 4-aminothiophenol was adopted as the bio-linkers to facilitate the antibody-antigen interaction. A more stable three-electrode configuration was miniaturized and laid out onto a biochip. A microfluidics subsystem based on opto-piezoelectronic technology was also integrated to create the microfluidic biochip system. To improve the detection sensitivity associated with the reduction in biochip size, electrochemistry simulation was used to investigate several potentially effective means. We found that the electric field on the edge near working electrode and counter electrode was higher, which was verified by using atomic force microscopy to measure the surface potential. With the successful verification, we explored the configuration, i.e., lengthened the edge of working electrode and counter electrode without changing the areas of working electrode and counter electrode and the gap between these two electrodes, so as to evaluate the possibility of improving the measurement efficiency in our newly developed biochips. Detailed design, simulation and experimental results, improved design identified, etc. were all presented in detail.

Hydrodynamic focusing is a powerful technique frequently used in microfluidics that presents a wide range of applications since it allows focusing the sample flowing in the device to a narrow region in the center of the microchannel. In fact thanks to the laminarity of the fluxes in microchannels it is possible to confine the sample solution with a low flow rate by using a sheath flow with a higher flow rate. This in turn allows the flowing of one sample element at a time in the detection region, thus enabling analysis on single particles. Femtosecond laser micromachining is ideally suited to fabricate device integrating full hydrodynamic focusing functionalities thanks to the intrinsic 3D nature of this technique, especially if compared to expensive and complicated lithographic multi-step fabrication processes. Furthermore, because of the possibility to fabricate optical waveguides with the same technology, it is possible to obtain compact optofluidic devices to perform optical analysis of the sample even at the single cell level, as is the case for optical cell stretchers and sorters. In this work we show the fabrication and the fluidic characterization of extremely compact devices having only two inlets for 2D (both in vertical and horizontal planes) as well as full 3D symmetric hydrodynamic focusing. In addition we prove one of the possible application of the hydrodynamic focusing module, by fabricating and validating (both with polystyrene beads and erythrocytes) a monolithic cell counter obtained by integrating optical waveguides in the 3D hydrodynamic focusing device.

In this paper, a simple and versatile method is presented which enables detection of nitric oxide (NO) released from vascular endothelial cells (ECs) cultured in microfluidic structures. The culturing system and NO measurement method allow cell shape to be controlled in a non-invasive manner using microfluidic structures while NO release is monitored for cell shape versus function studies. The culturing system consists of arrays of polydimethylsiloxane (PDMS) fluidic channels 120 micrometers in depth and ranging from 100 micrometers to 3 mm in width. The number of channels in each array is varied to yield a constant cell culture surface area (75 mm²) independent of channel width. The channel surfaces are collagen-coated and ECs are cultured to confluence within the channels. A cell scraper is then used to scrape extraneous cells cultured between channels, and NO measurements are made 18 to 24 hours later. A chemiluminescence-based sensor system (NOA 280i, Sievers NO Analyzer) is utilized to measure sample NO. Initial results indicate that NO concentrations can be measured from different microfluidic channel-containing samples using this method. It is shown that there is no significant difference in NO concentration derived from channels of different widths even though the degree of cell elongation varies due to physical constraint by microfluidic channel walls. However, cells treated with TNFα release more NO than untreated cells in fluidic channels, which is comparable to the function of ECs cultured in conventional culturing systems such as culturing dishes.

The concept of artificial pancreas, which comprises an insulin pump, a continuous glucose meter and a control algorithm, is a major step forward in managing patient with type 1 diabetes mellitus. The stability of the control algorithm is based on short-term precision micropump to deliver rapid-acting insulin and to specific integrated sensors able to monitor any failure leading to a loss of accuracy. Debiotech’s MEMS micropump, based on the membrane pump principle, is made of a stack of 3 silicon wafers. The pumping chamber comprises a pillar check-valve at the inlet, a pumping membrane which is actuated against stop limiters by a piezo cantilever, an anti-free-flow outlet valve and a pressure sensor. The micropump inlet is tightly connected to the insulin reservoir while the outlet is in direct communication with the patient skin via a cannula. To meet the requirement of a pump dedicated to closed-loop application for diabetes care, in addition to the well-controlled displacement of the pumping membrane, the high precision of the micropump is based on specific actuation profiles that balance effect of pump elasticity in low-consumption push-pull mode.

This paper describes a simple method of measuring refractive indices of liquids with a small hollow prism. When a He-Ne laser beam is sent to the liquid filled prism it is angularly deviated or deflected. If different liquids are injected in the prism, one at a time, the angular position of the emerging beam changes. These angular positions are detected by an optical fiber that samples a small area of the Gaussian beam cross section. A calibration curve relating intensity as a function of the liquid refractive index is obtained.

Recent advances in nanofabrication and computational electromagnetic design techniques have enabled the realization of metallic nanostructures in different shapes and sizes with adjustable resonance frequencies. To date, many metamaterial designs in various geometries with the used of different materials have been presented for the applications of surface plasmons, cloaking, biosensing, and frequency selective surfaces^1-5. Surface plasmons which are collective electron oscillations on metal surfaces ensure that plasmonic nanoantennas can be used in many applications like biosensing at infrared (IR) and visible regions. The nanostructure that we introduce has a unit cell that consists of Jerusalem crossshaped nanoaperture on a gold layer, which is standing on suspended SiN_x, Si or glass membranes. The proposed nanoaperture antenna array has a regular and stable spectral response. In this study, we present sensitivity of the resonance characteristics of Jerusalem cross-shaped nanoaperture antenna arrays to the changes in substrate parameters and metal thickness. We demonstrate that resonance frequency values can be adjusted by changing the thicknesses and types of the dielectric substrate and the metallic layer. Numerical calculations on spectral response of the nanoantenna array are performed by using Finite Difference Time Domain (FDTD) method⁶. The results of the simulations specify that resonance frequencies, the reflectance and transmittance values at resonances, and the band gap vary by the change of substrate parameters and metal thicknesses. These variations is a sign of that the proposed nanoantenna can be employed for sensing applications.

One key challenge in the field of microfluidics and lab-on-a-chip experiments for biological or chemical applications is the remote manipulation of fluids, droplets and particles. These can be volume elements of reactants, particles coated with markers, cells or many others. Light-driven microfluidics is one way of accomplishing this challenge. In our work, we manipulated micrometre sized polystyrene beads in a microfluidic environment by inducing thermal flows. Therefore, the beads were held statically in an unstructured microfluidic chamber, containing a dyed watery solution. Inside this chamber, the beads were moved along arbitrary trajectories on a micrometre scale. The experiments were performed, using a MOEMS (micro-opto-electro-mechanical-systems)-based laser scanner with a variable focal length. This scanner system is integrated in a compact device, which is flexibly applicable to various microscope setups. The device utilizes a novel approach for varying the focal length, using an electrically tunable lens. A quasi statically driven MOEMS mirror is used for beam steering. The combination of a tunable lens and a dual axis micromirror makes the device very compact and robust and is capable of positioning the laser focus at any arbitrary location within a three dimensional working space. Hence, the developed device constitutes a valuable extension to manually executed microfluidic lab-on-chip experiments.