In this paper we outline some of the manufacturing advantages and challenges of working with thermoplastic elastomers as an alternative to traditional polydimethysiloxane (PDMS) for flexible and reversibly bonded microfluidic systems. Unlike PDMS, thermoplastic elastomers can be processed with many industrial polymer manufacturing technologies such as extrusion, injection molding, hot embossing and others, potentially permitting much more scalable production and cheaper costs per part. Unlike a more rigid thermoplastic, these thermoplastic elastomers are typically much easier to bond, either reversibly or permanently due to their inherent compliance and subsequent low pressures necessary to seal channels and reservoirs. Unlike PDMS however, where one material (Sylgard 184) dominates the literature, there have not been many in depth investigations into thermoplastic elastomers and their relative performance and applicability to microfabrication. We show a comparison between several categories of thermoplastic elastomer to demonstrate what issues may be encountered and to demonstrate that even for labs with minimal equipment, academic prototyping with these materials is not necessarily any more challenging than PDMS.

In academia the rapid and flexible creation of microfluidic chips is of great importance for microfluidic research. Besides polymers glass is a very important material especially when high chemical and temperature resistance are required. However, glass structuring is a very hazardous process which is not accessible to most members of the microfluidic community. We therefore sought a new method for the rapid and simple creation of transparent microfluidic glass chips by structuring and sintering amorphous silica suspensions. The whole process from a digital mask layout to a microstructured glass sheet can be done within two days. In this paper we show the applicability of this process to fabricate capillary driven microfluidic systems.

There are two main challenges in producing a robust, paper-based analytical device. The first one is to create a hydrophobic barrier which unlike the commonly used wax barriers does not break if the paper is bent. The second one is the creation of the (bio-)specific sensing layer. For this proteins have to be immobilized without diminishing their activity. We solve both problems using light-based fabrication methods that enable fast, efficient manufacturing of paper-based analytical devices. The first technique relies on silanization by which we create a flexible hydrophobic barrier made of dimethoxydimethylsilane. The second technique demonstrated within this paper uses photobleaching to immobilize proteins by means of maskless projection lithography. Both techniques have been tested on a classical lithography setup using printed toner masks and on a lithography system for maskless lithography. Using these setups we could demonstrate that the proposed manufacturing techniques can be carried out at low costs. The resolution of the paper-based analytical devices obtained with static masks was lower due to the lower mask resolution. Better results were obtained using advanced lithography equipment. By doing so we demonstrated, that our technique enables fabrication of effective hydrophobic boundary layers with a thickness of only 342 μm. Furthermore we showed that flourescine-5-biotin can be immobilized on the non-structured paper and be employed for the detection of streptavidinalkaline phosphatase. By carrying out this assay on a paper-based analytical device which had been structured using the silanization technique we proofed biological compatibility of the suggested patterning technique.

We present two different platforms integrating silicon photonic biosensors. One is based on integration with reaction tubes to be compatible with traditional lab approaches. The other uses through-chip fluidics in order to achieve better mixing of the analyte.

The demand for rapid screening technologies, to be used outside of a traditional healthcare setting, has been vastly expanding. This is requiring a new engineering platform for faster and cost effective techniques to be easily adopted through forward-thinking manufacturing procedures, i.e., advanced miniaturisation and heterogeneous integration of high performance microfluidics based point-of-care testing (POCT) systems. Although there has been a considerable amount of research into POCT systems, there exist tremendous challenges and bottlenecks in the design and manufacturing in order to reach a clinical acceptability of sensitivity and selectivity, as well as smart microsystems for healthcare. The project aims to research how to enable scalable production of such complex systems through 1) advanced miniaturisation of a physical layout and opto-electronic component allocation through an optimal design; and 2) heterogeneous integration of multiplexed fluorescence detection (MFD) for in vitro POCT. Verification is being arranged through experimental testing with a series of dilutions of commonly used fluorescence dye, i.e. Cy5. Iterative procedures will be engaged until satisfaction of the detection limit, of Cy5 dye, 1.209x10^-10 M. The research creates a new avenue of rapid screening POCT manufacturing solutions with a particular view on high performance and multifunctional detection systems not only in POCT, but also life sciences and environmental applications.

Economically advantageous microfabrication technologies for lab-on-a-chip diagnostic devices substituting commonly used glass etching or injection molding processes are one of the key enablers for the emerging market of microfluidic devices. On-site detection in fields of life sciences, point of care diagnostics and environmental analysis requires compact, disposable and highly functionalized systems. Roll-to-roll production as a high volume process has become the emerging fabrication technology for integrated, complex high technology products within recent years (e.g. fuel cells). Differently functionalized polymer films enable researchers to create a new generation of lab-on-a-chip devices by combining electronic, microfluidic and optical functions in multilayer architecture. For replication of microfluidic and optical functions via roll-to-roll production process competitive approaches are available. One of them is to imprint fluidic channels and optical structures of micro- or nanometer scale from embossing rollers into ultraviolet (UV) curable lacquers on polymer substrates. Depending on dimension, shape and quantity of those structures there are alternative manufacturing technologies for the embossing roller. Ultra-precise diamond turning, electroforming or casting polymer materials are used either for direct structuring or manufacturing of roller sleeves. Mastering methods are selected for application considering replication quality required and structure complexity. Criteria for the replication quality are surface roughness and contour accuracy. Structure complexity is evaluated by shapes producible (e.g. linear, circular) and aspect ratio. Costs for the mastering process and structure lifetime are major cost factors. The alternative replication approaches are introduced and analyzed corresponding to the criteria presented. Advantages and drawbacks of each technology are discussed and exemplary applications are presented.

We report an optical fiber chemical sensor based on a focused ion beam processed optical fiber. The demonstrated sensor is based on a cavity formed onto a standard 1550 nm single-mode fiber by either chemical etching, focused ion beam milling (FIB) or femtosecond laser ablation, on which side channels are drilled by either ion beam milling or femtosecond laser irradiation. The encapsulation of the cavity is achieved by optimized fusion splicing onto a standard single or multimode fiber. The empty cavity can be used as semi-curved Fabry-Pérot resonator for gas or liquid sensing. Increased reflectivity of the formed cavity mirrors can be achieved with atomic layer deposition (ALD) of alternating metal oxides. For chemical selective optical sensors, we demonstrate the same FIB-formed cavity concept, but filled with different materials, such as polydimethylsiloxane (PDMS), poly(methyl methacrylate) (PMMA) which show selective swelling when immersed in different solvents. Finally, a reducing agent sensor based on a FIB formed cavity partially sealed by fusion splicing and coated with a thin ZnO layer by ALD is presented and the results discussed. Sensor interrogation is achieved with spectral or multi-channel intensity measurements.

Microfluidic technologies have largely been realized within enclosed microchannels. While powerful, a principle limitation of closed-channel microfluidics is the difficulty for sample extraction and downstream processing. To address this limitation and expand the utility of microfluidic analytical separation tools, we developed an openchannel hydrogel architecture for rapid protein analysis. Designed for compatibility with slab-gel polyacrylamide gel electrophoresis (PAGE) reagents and instruments, we detail the development of free-standing polyacrylamide gel (fsPAG) microstructures supporting electrophoretic performance rivalling that of microfluidic platforms. Owing to its open architecture – the platform can be easily interfaced with automated robotic controllers and downstream processing (e.g., sample spotters, immunological probing, mass spectroscopy). The fsPAG devices are directly photopatterened atop of and covalently attached to planar polymer or glass surfaces. Due to the fast < 1 hr design-prototype-test cycle – significantly faster than mold based fabrication techniques – rapid prototyping devices with fsPAG microstructures provides researchers a powerful tool for developing custom analytical assays. Leveraging the rapid prototyping benefits - we up-scale from a unit separation to an array of 96 concurrent fsPAGE assays in 10 min run time driven by one electrode pair. The fsPAGE platform is uniquely well-suited for massively parallelized proteomics, a major unrealized goal from bioanalytical technology.

Previously we modeled the theoretical benefits of using microfluidic channels that utilize a “Cathedral Chamber” design, in which the ceiling is supported by an array of periodic posts, compared to an array of parallel microfluidic channels. We developed a semi-automated technique that combines a rule-base defect placement system with a 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 rate. Earlier results indicate that Cathedral Chambers, that are supported by an array of 10 by 11 periodic posts with the same size as the spacing have six times higher lifetime expectancy compared to an array of 10 parallel channels, likely due to the provision of multiple paths during localized blockage formation in the Cathedral Chamber. In this paper, we have expanded our investigations by considering the defect tolerance sensitivity to scale by altering parameters such as the number and size of the posts and overall size of the chamber. For one set of simulations, we used the same number of posts in the chamber and the same starting position for the first 10 blockages as in our previous work. However, we shrank the size of the posts to 66% of their former size so that the new channels (flow pathways) are twice the size of the modified posts. In addition, we have also performed initial simulations based on wider microfluidic channels supported by an array of 20 by 11 periodic posts in order to explore their microfluidic behavior and lifetime.

Lorentz force is the pumping basis of many electromagnetic micropumps used in lab-on-a-chip. In this paper a novel reciprocating single-chamber micropump is proposed, in which the actuation technique is based on Lorentz force acting on an array of microwires attached on a membrane surface. An alternating current is applied through the microwires in the presence of a magnetic field. The resultant force causes the membrane to oscillate and pushes the fluid to flow through microchannel using a ball-valve. The pump chamber (3 mm depth) was fabricated on a Polymethylmethacrylate (PMMA) substrate using laser engraving technique. The chamber was covered by a 60 μm thick hyper-elastic latex rubber diaphragm. Two miniature permanent magnets capable of providing magnetic field of 0.09 T at the center of the diaphragm were mounted on each side of the chamber. Square wave electric current with low-frequencies was generated using a function generator. Cylindrical copper microwires (250 μm diameter and 5 mm length) were attached side-by-side on top surface of the diaphragm. Thin loosely attached wires were used as connectors to energize the electrodes. Due to large displacement length of the diaphragm (~3 mm) a high efficiency (~90%) ball valve (2 mm diameter stainless steel ball in a tapered tubing structure) was used in the pump outlet. The micropump exhibits a flow rate as high as 490 μl/s and pressure up to 1.5 kPa showing that the pump is categorized among high-flow-rate mechanical micropumps.

In this paper, we present a new type of piezoelectric composite material, optopiezoelectric thin-film, to serve as a lightactivated micropump for integrating with a microfluidic device. By using a photoconductive material (titanium oxide phthalocyanine) to serve as one of the electrodes of a piezoelectric polyvinylidene fluoride (PVDF) polymer, multiple locations of this optopiezoelectric thin-film can be actuated independently with one driving voltage source and a programmable light mask. Integrating this optopiezoelectric thin-film to a microfluidic device, complex operations of a multi-functioned microfluidic device can potentially be simplified and scaled up. Here, we present our preliminary result to demonstrate the feasibility of using one optopiezoelectric thin-film to serve as two microfluidic micropumps controlled by a light mask.

Electrokinetics has many applications in a wide range of areas, such as lab-on-a-chip and biomedical microdevices. The electrothermal effect has been used for biofluid delivery systems since it has high pumping efficiency for high conductive liquids (>0.1 S/m) compared to other electrokinetic techniques such as electroosmosis. AC electrothermal (ACET) micropumps are based on the temperature gradient caused by Joule heating or an external heat source, which generates permittivity and conductivity gradients in the bulk of the liquid. When the liquid is subjected to an electric field, the ACET force is created. Electrode geometry significantly affects the electric field distribution, which can yield stronger ACET forces. Previously electrode dimension optimization has been performed for a single-array coplanar asymmetric configuration in order to obtain maximum ACET velocities. In this paper we expand the study to other governing parameters in a multiple-row microelectrode array configuration consisting of microelectrodes placed on top, bottom, and/or side walls of a microchannel. The studied parameters are the substrate material and thickness, ambient temperature, fluid viscosity, and actuation frequency. Electrode dimensions remain constant during the study (120 μm wide and 20 μm thin electrodes, 20 μm gap). The study is performed using finite element analysis software for one pair of microelectrodes on each array with periodic boundary conditions. The simulation data is then compared with experimental data for a single combination of the aforementioned parameters. The results show that the effect of these parameters on ACET flow can be significant.

The 3D laser direct-writing technology is aimed at the modeling of arbitrary three-dimensional (3D) complex microstructures by scanning a laser-focusing point along predetermined trajectories. Through the perspective technique, the details of designed 3D structures can be properly fabricated in a microchannel. This study introduces a direct reading flow meter and a 3D passive mixer fabricated by laser direct writing for microfluidic applications. The flow meter consists of two rod-shaped springs, a pillar, an anchor, and a wedge-shaped indicator, installed inside a microfluidic channel. The indicator is deflected by the flowing fluid while restrained by the spring to establish an equilibrium indication according to the flow rate. The measurement is readily carried out by optical microscopy observation. The 3D passive Archimedes-screw-shaped mixer is designed to disturb the laminar flow 3D direction for enhancing the mixing efficiency. The simulation results indicate that the screw provides 3D disturbance of streamlines in the microchannel. The mixing demonstration for fluids flowing in the micrchannel approximately agrees with the simulation result. Thanks to the advantage of the laser direct writing technology, this study performs the ingenious applications of 3D structures for microchannels.

DNA sequencing in a lab-on-a-chip aims at providing cheap, high-speed analysis of low reagent volumes to, e.g., identify genomic deletions or insertions associated with genetic illnesses. Detecting single base-pair insertions/deletions from DNA fragments in the diagnostically relevant range of 150−1000 base-pairs requires a sizing accuracy of S < 10^-3. Here we demonstrate S = 4×10^-4. A microfluidic chip was post-processed by femtosecond-laser writing of an optical waveguide. 12 blue-labeled and 23 red-labeled DNA fragments were separated in size by capillary electrophoresis, each set excited by either of two lasers power-modulated at different frequencies, their fluorescence detected by a photomultiplier, and blue/red signals distinguished by Fourier analysis. Different calibration strategies were tested: a) use either set of DNA molecules as reference to calibrate the set-up and identify the base-pair sizes of the other set in the same flow experiment, thereby eliminating variations in temperature, wall-coating and sieving-gel conditions, and actuation voltages; b) use the same molecular set as reference and sample with the same fluorescence label, flown in consecutive experiments; c) perform cross-experiments based on different molecular sets with different labels, flown in consecutive experiments. From the results we conclude: Applying quadratic instead of linear fit functions improves the calibration accuracy. Blue-labeled molecules are separated with higher accuracy. The influence of dye label is higher than fluctuations between two experiments. Choosing a single, suitable dye label combined with reference calibration and sample investigation in consecutive experiments results in S = 4×10^-4, enabling detection of single base-pair insertion/deletion in a lab-on-a-chip.

We present a novel microfluidic surface-enhanced Raman scattering (SERS) sensor for rapid and label-free biomolecular detection. Our sensor design mitigates a common limiting factor in microfluidic SERS sensors that utilize integrated nanostructures: low-efficiency transport of biomolecules to nanostructured surface which adversely impacts sensitivity. Our strategy is to increase the total usable nanostructured surface area, which provides more adsorption sites for biomolecules. Specifically, nanoporous gold disk (NPGD) array, a highly effective SERS substrate, has been monolithically integrated inside a microfluidic chip. Individual NPGD is known to feature an order of magnitude larger surface area than its projected disk area. The increased surface area arises from nanoscale pores and ligaments 3- dimensionally distributed in the NPGD, which manifest themselves as high-density SERS hot-spots. High-density NPGD arrays further guarantee large coverage of these hot-spots on the microchannel floor. The SERS-active NPGD arrays enable highly-reproducible SERS measurements with relative intensity variations from 8% to -8%. R6G solutions in the concentrations ranging from 1 μM to 1 mM have been detected and quantitatively evaluated, and the performance of the sensor in continuous-flow condition has been assessed. Moreover, the sensor’s capabilities have been studied by detecting and identifying a physiological metabolite (urea), and the results show lower detection limit compared to best results from most recent work using integrated nanostructured surface inside microchannels. We expect that the sensor would be applicable for detecting, identifying and quantifying molecules for some point-of-care applications, i.e. urine screening.

For cancer diagnoses, core biopsies (CBs) obtained from patients using coring needles (CNs) are traditionally visualized and assessed on microscope slides by pathologists after samples are processed and sectioned. A fundamental gain in optical information (i.e., diagnosis/staging) may be achieved when whole, unsectioned CBs (L = 5-20, D = 0.5-2.0 mm) are analyzed in 3D. This approach preserves CBs for traditional pathology and maximizes the diagnostic potential of patient samples. To bridge CNs/CBs with imaging, our group developed a microfluidic device that performs biospecimen preparation on unsectioned CBs for pathology. The ultimate goal is an automated and rapid point-of-care system that aids pathologists by processing tissue for advanced 3D imaging platforms. An inherent, but essential device feature is the microfluidic transport of CBs, which has not been previously investigated. Early experiments demonstrated proof-of-concept: pancreas CBs (D = 0.3-2.0 mm) of set lengths were transported in straight/curved microchannels, but dimensional tolerance and flow rates were variable, and preservation of CB integrity was uncontrolled. A second study used metal cylinder substitutes (L = 10, D = 1 mm) in microchannels to understand the transport mechanism. However, CBs are imperfectly shaped, rough, porous and viscoelastic. In this study, fresh/formalin-fixed porcine and human pancreas CBs were deposited into our device through a custom interface using clinical CNs. CB integrity (i.e., sample viability) may be assessed at every stage using an optomechanical metric: physical breaks were determined when specimen intensity profile data deviated beyond x_avg + 2σ. Flow rates for human CBs were determined for several CNs, and microfluidic transport of fresh and formalin-fixed CBs was analyzed.

Aneurysms are pockets of blood that collect outside blood vessel walls forming dilatations and leaving arterial walls very prone to rupture. There is little information concerning the causes of intracranial aneurysm formation, growth, and rupture. Current treatments include: (1) clipping, and (2) coil embolization, including stent-assisted coiling. Further, the evolution of any aneurysm is assumed to be caused by the remodeling of the affected blood vessel’s material constituents (tunica intima, tunica media, or tunica adventitia). Velocity, pressure, and wall shear stresses aid in the disease development of aneurysmal growth, while the shear force mechanisms effecting wound closure are elusive. To study aneurysm pathogenesis, a lab-on-a-chip device is the key to discovering the underlying mechanisms of these lesions. A two-dimensional microfluidic model, the Aneurysm-on-a-Chip™ (AOC), was the logical answer to study particle flow within an aneurysm “sac”. The AOC apparatus can track particles/cells when it is coupled to particle image velocimetry software (PIV) package. The AOC fluid flow was visualized using standard microscopy techniques with commercial microparticles and human aortic smooth muscle cells (HASMC). Images were taken during fluid flow experiments and PIV was utilized to monitor the flow of particles within the “sac” region, as well as particles entering and exiting the device. Quiver plots were generated from fluid flow experiments using standard 7 μm latex particles and fixed HASMC in PBS. PIV analysis shows that the particles flowed nicely from input to output. Wall shear stress provided evidence that there was some back flow at the edges of the “sac” – an indicator of aneurysm development in human patients.

Single cell analyses of RNA and DNA are crucial to understanding the heterogeneity of cell populations. The numbers of approaches to single cells analyses are expanding, but sequence specific measurements of nucleic acids have been mostly limited to studies of either DNA or RNA, and not both. This remains a challenge as RNA and DNA have very similar physical and biochemical properties, and cross-contamination with each other can introduce false positive results. We present an electrokinetic technique which creates the opportunity to fractionate and deliver cytoplasmic RNA and genomic DNA to independent downstream analyses. Our technique uses an on-chip system that enables selective lysing of cytoplasmic membrane, extraction of RNA (away from genomic DNA and nucleus), focusing, absolute quantification of cytoplasmic RNA mass. The absolute RNA mass quantification is performed using fluorescence observation without enzymatic amplification in < 5 min. The cell nucleus is left intact and the relative genomic DNA amount in the nucleus can be measured. We demonstrate the technique using single mouse B lymphocyte cells, for which we extracted an average of 14.1 pg total cytoplasmic RNA per cell. We also demonstrate correlation analysis between the absolute amount of cytoplasmic RNA and relative amount of genomic DNA, showing heterogeneity associated with cell cycle.

We have previously addressed experimentally blood fluidodynamics in microcapillaries by coupling optical microscopy to pixelated detection. By computing the Cross-Correlation Function (CCF) of signals coming from pixels at a distance along the flow we obtained information on the flow speed and direction. The extension of these experiments to more complex systems with high branching of capillaries and/or inverted flows needs a theoretical investigation that we present here. We focus first on straight capillaries and harmonic flows between a minimum V_min ≠ 0 and a maximum V_max flow speed. The CCF shows multiple peaks at lag times that correspond closely to the maximum and minimum flow speeds. The general analytical expression of the CCF is given, the position of its maxima are discussed by means of geometrical considerations and numerical analysis and an experimental validation are presented. The second case that we study is the flow in the branches of a y-shaped junction in a microcapillary. By simply modeling the branching in laminar flow (low Reynold numbers) and assuming a smooth transition of speeds along the branches we derive a simple numerical model to compute the trajectories of micro-beads. We estimate the flow speed in the branches by computing the CCFs between linear regions of interest set perpendicular to the axes of the branches.

Urinary tract infections (UTIs) are among the most common bacterial infections and pose a significant healthcare burden. The growing trend in antibiotic resistance makes it mandatory to develop diagnostic kits which allow not only the determination of a pathogen but also the antibiotic resistances. We have developed a microfluidic cartridge which takes a direct urine sample, extracts the DNA, performs an amplification using batch-PCR and flows the sample over a microarray which is printed into a microchannel for fluorescence detection. The cartridge is injection-molded out of COP and contains a set of two-component injection-molded rotary valves to switch between input and to isolate the PCR chamber during thermocycling. The hybridization probes were spotted directly onto a functionalized section of the outlet microchannel. We have been able to successfully perform PCR of E.coli in urine in this chip and perform a fluorescence detection of PCR products. An upgraded design of the cartridge contains the buffers and reagents in blisters stored on the chip.

It is estimated that about one-third of the world’s population has already been infected by tuberculosis. Mycobacterium tuberculosis, in general, can result in an active case of tuberculosis in approximately 5%-10% of those who suffer from latent tuberculosis and the chance of becoming ill is the highest within one of year of getting the disease. Although a newly developed methods called interferon gamma release assay (IGRA) can monitor CD4 cells secreted cytokine to diagnose tuberculosis (TB) condition. However, it is difficult to count total numbers of cytokine secreted CD4 cells, which make the diagnosis less accurate. Therefore, we develop a functionalized polydimethylsiloxane (PDMS) device using glutaraldehyde to capture CD4 cells. To enhance the capture efficiency, we use COMSOL simulation to optimize the arrangement of PDMS micro pillars to make cells uniformly distributed in the device. Our preliminary data showed the microfluidic configuration in a circular shape with HCP patterned micro pillars turned 30 degrees offers the highest cell capture rate.

When using appropriate materials and microfabrication techniques, with the small dimensions the mechanical stability of microstructured devices allows for processes at high pressures without loss in safety. The largest area of applications has been demonstrated in green chemistry and bioprocesses, where extraction, synthesis and analyses often excel at high densities and high temperatures. This is accessible through high pressures. Capillary chemistry has been used since long but, just like in low-pressure applications, there are several potential advantages in using microfluidic platforms, e.g., planar isothermal set-ups, large local variations in geometries, dense form factors, small dead volumes and precisely positioned microstructures for control of reactions, catalysis, mixing and separation. Other potential applications are in, e.g., microhydraulics, exploration, gas driven vehicles, and high-pressure science. From a review of the state-of-art and frontiers of high pressure microfluidics, the focus will be on different solutions demonstrated for microfluidic handling at high pressures and challenges that remain.

The continuous monitoring of the environment for lethal pathogens is a central task in the field of biothreat detection. Typical scenarios involve air-sampling in locations such as public transport systems or large public events and a subsequent analysis of the samples by a portable instrument. Lab-on-a-chip technologies are one of the promising technological candidates for such a system. We have developed an integrated microfluidic system with automatic sampling for the detection of CBRNE-related pathogens. The chip contains a two-pronged analysis strategy, on the one hand an immunological track using antibodies immobilized on a frit and a subsequent photometric detection, on the other hand a molecular biology approach using continuous-flow PCR with a fluorescence end-point detection. The cartridge contains two-component molded rotary valve to allow active fluid control and switching between channels. The accompanying instrument contains all elements for fluidic and valve actuation, thermal control, as well as the two detection modalities. Reagents are stored in dedicated reagent packs which are connected directly to the cartridge. With this system, we have been able to demonstrate the detection of a variety of pathogen species.

Imaging cells in a microfluidic chamber with an area scan camera is difficult due to motion blur and data loss during frame readout causing discontinuity of data acquisition as cells move at relatively high speeds through the chamber. We have developed a method to continuously acquire high-resolution images of cells in motion through a microfluidics chamber using a high-speed line scan camera. The sensor acquires images in a line-by-line fashion in order to continuously image moving objects without motion blur. The optical setup comprises an epi-illuminated microscope with a 40X oil immersion, 1.4 NA objective and a 150 mm tube lens focused on a microfluidic channel. Samples containing suspended cells fluorescently stained with 0.01% (w/v) proflavine in saline are introduced into the microfluidics chamber via a syringe pump; illumination is provided by a blue LED (455 nm). Images were taken of samples at the focal plane using an ELiiXA+ 8k/4k monochrome line-scan camera at a line rate of up to 40 kHz. The system’s line rate and fluid velocity are tightly controlled to reduce image distortion and are validated using fluorescent microspheres. Image acquisition was controlled via MATLAB’s Image Acquisition toolbox. Data sets comprise discrete images of every detectable cell which may be subsequently mined for morphological statistics and definable features by a custom texture analysis algorithm. This high-throughput screening method, comparable to cell counting by flow cytometry, provided efficient examination including counting, classification, and differentiation of saliva, blood, and cultured human cancer cells.

Development of efficient methods for isolation and manipulation of microorganisms is essential to study unidentified and yet-to-be cultured microbes originating from a variety of environments. The discovery of novel microbes and their products have the potential to contribute to the development of new medicines and other industrially important bioactive compounds. In this paper we describe the design, fabrication and validation of an optofluidic device capable of redirecting microbes within a flow using optical forces. The device holds promise to enable the high throughput isolation of single microbes for downstream culture and analysis. Optofluidic devices are widely used in clinical research, cell biology and biomedical engineering as they are capable of performing analytical functions such as controlled transportation, compact and rapid processing of nanolitres to millilitres of clinical or biological samples. We have designed and fabricated a three dimensional optofluidic device to control and manipulate microorganisms within a microfluidic channel. The device was fabricated in fused silica by ultrafast laser inscription (ULI) followed by selective chemical etching. The unique three-dimensional capability of ULI is utilized to integrate microfluidic channels and waveguides within the same substrate. The main microfluidic channel in the device constitutes the path of the sample. Optical waveguides are fabricated at right angles to the main microfluidic channel. The potential of the optical scattering force to control and manipulate microorganisms is discussed in this paper. A 980 nm continuous wave (CW) laser source, coupled to the waveguide, is used to exert radiation pressure on the particle and particle migrations at different flow velocities are recorded. As a first demonstration, device functionality is validated using fluorescent microbeads and initial trials with microalgae are presented.

Electrowetting has been widely studied for various optical applications such as optical switch, sensor, prism, and display. In this study, vari-focal liquid lens array is developed using electrowetting principle to construct integral 3-dimensional imaging. The electrowetting principle that changes the surface tension by applying voltage has several advantages to realize active optical device such as fast response time, low electrical consumption, and no mechanical moving parts. Two immiscible liquids that are water and oil are used for forming lens. By applying a voltage to the water, the focal length of the lens could be tuned as changing contact angle of water. The fabricated electrowetting vari-focal liquid lens array has 1mm diameter spherical lens shape that has 1.6mm distance between each lens. The number of lenses on the panel is 23x23 and the focal length of the lens array is simultaneously tuned from -125 to 110 diopters depending on the applied voltage. The fabricated lens array is implemented to integral 3-dimensional imaging. A 3D object is reconstructed by fabricated liquid lens array with 23x23 elemental images that are generated by 3D max tools. When liquid lens array is tuned as convex state. From vari-focal liquid lens array implemented integral imaging system, we expect that depth enhanced integral imaging can be realized in the near future.

Integrating photonic waveguide sensors with microfluidics is promising in achieving high-sensitivity and cost-effective biological and chemical sensing applications. One challenge in the integration is that an air gap would exist between the microfluidic channel and the photonic waveguide when the micro-channel and the waveguide intersect. The air gap creates a path for the fluid to leak out of the micro-channel. Potential solutions, such as oxide deposition followed by surface planarization, would introduce additional fabrication steps and thus are ineffective in cost. Here we propose a reliable and efficient approach for achieving closed microfluidic channels on a waveguide sensing chip. The core of the employed technique is to add waveguide crossings, i.e., perpendicularly intersecting waveguides, to block the etched trenches and prevent the fluid from leaking through the air gap. The waveguide crossings offer a smooth interface for microfluidic channel bonding while bring negligible additional propagation loss (0.024 dB/crossing based on simulation). They are also efficient in fabrication, which are patterned and fabricated in the same step with waveguides. We experimentally integrated microfluidic channels with photonic crystal (PC) microcavity sensor chips on silicon-on-insulator substrate and demonstrated leak-free sensing measurement with waveguide crossings. The microfluidic channel was made from polydimethylsiloxane (PDMS) and pressure bonded to the silicon chip. The tested flow rates can be varied from 0.2 μL/min to 200 μL/min. Strong resonances from the PC cavity were observed from the transmission spectra. The spectra also show that the waveguide crossings did not induce any significant additional loss or alter the resonances.

Electrokinetic micropumps have been widely used in lab-on-a-chip devices. The AC electrothermal (ACET) effect is highly efficient for biofluidic micropumping, but is unable to generate high flow rates. Attempts to increase ACET flows, such as applying a wide range of actuation voltages, using asymmetric microelectrode arrays and using 3D microelectrodes have been reported. In this paper a novel idea of employing circular coplanar asymmetric microelectrodes placed on the perimeter of a microchannel is explored. An array of microelectrodes is simulated using COMSOL Multiphysics software. The micropump output shows relatively high flow rates compared to other ACET micropumps which have the same electrode dimensions. Moreover, the idea of using different micropumps with scaled dimensions is investigated. The results show that a highly efficient ACET micropump can be achieved if an appropriate electrode size-to-channel dimension ratio is selected. The results also show that a micropump with a scale of 0.2 can show negligible flow rate, but if the electrodes are used in a micropump with the scale of 1, a flow rate of 15 ×10⁶ μm³/s can be generated. This new ACET pump design can be utilized for lab-on-a-chip applications, specifically in biofluid delivery systems.

The tumor microenvironment is a complex system which is not fully understood. New technologies are needed to provide a better understanding of the role of the tumor microenvironment in promoting metastasis. The Nano Intravital Device, or NANIVID, has been developed as an optically transparent, implantable tool to study the tumor microenvironment. Two etched glass substrates are sealed using a thin polymer membrane to create a reservoir with a single outlet. This reservoir is loaded with a custom hydrogel blend that contains selected factors for delivery to the tumor microenvironment. When the device is implanted in the tumor, the hydrogel swells and releases these entrapped molecules, forming a sustained concentration gradient. The NANIVID has previously been successful in manipulating the tumor microenvironment both in vitro as well as in vivo. As metastatic cells intravasate, it has been shown that some are able to do so unscathed and reach their new location, while others are cleaved during the process¹. There appears to be a correlation between cell migration and the mechanical properties of these cells. It is believed that these properties can be detected in real time by atomic force microscopy. In this study, metastatic MTLn3 rat mammary cells are seeded onto 1-dimensional microfibers and directed up a stable gradient of growth factor. The NANIVID device is placed behind our AFM tip, where it generates a stable chemotactic gradient of epidermal growth factor. Scanning confocal laser microscopy is also used to monitor movement of the cells over time. This experiment will shed light on the mechanical changes in metastatic cells as they undergo directed migration.

Microbubbles are widely used in many industries such as water treatment, drug coating, and ultrasonic contrast agents. Cross-flow focusing and co-flow focusing are considered basic mechanisms used for microbubble generation. Typically, to achieve micron-sized droplets requires structure dimensions in the same order of magnitude of the desired droplet sizes. In this paper we report a method of applying an external vibration to a cross-flow and co-flow focusing structure, which allows for smaller droplets to be generated. The junction dimension was 700×400 μm, and the channel width was 800 μm. The two assumed fluids are selected in a way that the Capillary number is high (Ca>10) to make use of necking effect occurred in the downstream. Linear vibration was exerted on the microchannel structure in the direction of central flow. A 2D structure was simulated using finite element software, and the numerical approach was then verified by comparing the experimental data of a typical cross-flow focusing structure taken from our previous study with the corresponding simulation assuming the same parameters. The results show that although the droplet generation regime depends on flow ratio (Q_a/Q_w) and vibration parameter (ampl×freq), Capillary number also has a significant effect on the regime. Briefly, applying a low-cost linear vibration to the conventional flow focusing structures can be used as an accurate controlling technique for increasing the chance of droplet generation. In fact, vibration motion can change the flow regime and breakup mechanism. It can also change the breakup point at which the droplets are formed.

Electrokinetics have been widely used in lab-on-a-chip devices for fluid manipulation applications. The AC electrothermal (ACET) effect is a highly efficient technique for biofluids (σ>0.1 S/m) active micromixing, which can be used in chemical, biological, and medical analysis systems. In this paper, a novel idea of employing microelectrode arrays placed on sidewalls of a fluidic microchannel for increasing the mixing efficiency of biofluids is numerically investigated. It was reported that coplanar asymmetric microelectrode arrays are capable of creating ACET vortices in the bulk of a high conductive electrolyte solution. Two electrode arrays can be placed on the sidewalls of a microchannel, each of which has a different role, one pumps the biofluid while the other mixes it. Two different actuation patterns were applied to the electrodes. One pair of microelectrodes was simulated and the simulation procedure was then verified by conducting experiments for ACET flow measurement in a similar geometry. Microelectrode arrays were fabricated on 1mm thick glass substrates using photolithography. A 800 μm thick fluidic microchannel was fabricated by soft lithography of Polydimethylsiloxane (PDMS). The results showed that such a technique can dramatically increase the mixing of the solution while pumping is taking place. The mechanism was capable of efficiently mixing biofluid solutions (resultant concentration ratio of up to 80%) in a short time (<3 min) and short distance (<600 μm) for a 300×300 μm2 fluidic microchannel cross section area. Medical analysis such as heterogeneous immunoassays can be potential applications of such micromixing technique.

This PDF contains Front Matter for volume 9320. This includes the Title Page, Table of Contents, and Conference Committee.