Integrated Lithographic Molding for Microneedle-Based Devices

This paper presents a new fabrication method consisting of lithographically defining multiple layers of high aspect-ratio photoresist onto preprocessed silicon substrates and release of the polymer by the lost mold or sacrificial layer technique, coined by us as lithographic molding. The process methodology was demonstrated fabricating out-of-plane polymeric hollow microneedles. First, the fabrication of needle tips was demonstrated for polymeric microneedles with an outer diameter of 250 mum, through-hole capillaries of 75-mum diameter and a needle shaft length of 430 mum by lithographic processing of SU-8 onto simple v-grooves. Second, the technique was extended to gain more freedom in tip shape design, needle shaft length and use of filling materials. A novel combination of silicon dry and wet etching is introduced that allows highly accurate and repetitive lithographic molding of a complex shape. Both techniques consent to the lithographic integration of microfluidic back plates forming a patch-type device. These microneedle-integrated patches offer a feasible solution for medical applications that demand an easy to use point-of-care sample collector, for example, in blood diagnostics for lithium therapy. Although microchip capillary electrophoresis glass devices were addressed earlier, here, we show for the first time the complete diagnostic method based on microneedles made from SU-8

Design of a microfabricated device for Ligase Detection Reaction (LDR)

The Ligase Detection Reaction (LDR) is a mutation detection technique used to identify point mutations in deoxyribonucleic acid (DNA). Developed by Francis Barany and associates at Cornell University it is used to find specific low abundant point mutations that may lead to colorectal cancer in the early stages of disease development. The research objective was to design and manufacture a microscale Ligase Detection Reaction (LDR) device in polycarbonate. The LDR module will be incorporated with other microdevices such as: Continuous Flow Polymerase Chain Reaction (CFRCR) and Capillary Electrophoresis (CE) in modular lab-on-a-chip technology. In making the microdevice, the duration of original reaction had to be scaled down from the current 2½ hours for 20 cycles for the macroscale reaction. It was found that an excess of primers in relation to PCR product was needed for efficient ligation. By changing the concentrations, volumes and time for the process the current time is down to 40 minutes for 20 cycles with indications that further time reductions are possible on the microscale. There are two mixing stages involved in the reaction. Micromixers were simulated in Fluent (v5.4, Lebanon, NH) and several test geometries selected for fabrication. Passive diffusion mixing was used based on obtaining high aspect ratios, 7 to 20. The mixers were made by SU-8 lithography, LIGA, laser ablation, and micromilling to characterize each fabrication method. It was found that LIGA was best for making the micromixers, but was the longest process. The micromixers are fabricated and tested using chemi-luminescence technique. For a successful reaction, temperatures of 0°C, 95°C and 65°C were needed. A stationary chamber was used for thermal cycling in which the sample sits while the temperature is cycled. Finite element analysis showed uniform temperatures in the rectangular 1.5μl chambers and that air slits can effectively separate the thermal cycle zone from the 0°C cooling zone and also isolate the mixing region. A test device was laid out and micromilled with the temperature zones maintained and fluid flow controlled. A commercial thin film heater and a thermoelectric module were used with PID controls to obtain the required process temperatures. Heating from 65°C to 95°C took 10 seconds, while cooling from 95°C to 65°C also took 10 seconds. The residence times at the required temperatures can be adapted to changes in the LDR

30 years of microfluidics

Microfluidics provides a great opportunity to create devices capable of outperforming classical techniques in biomedical and chemical research. In this review, the origins of this emerging field in the microelectronics industry are detailed. We also appraise how factors such as government funding influenced the development of new materials and fabrication techniques. Current applications of microfluidics are also examined and we highlight areas where work should be focussed in the future to ensure that the technology realises its full potential

Multiphase flows in polymer microfluidic systems

Continuous delivery of segmented reagents using pressure-driven multiphase flow in microchannels is a promising technology for high throughput microfluidic bioassays. Separation and encapsulation of the target reagents with another inert fluid provide many advantages over single phase flow in microfluidic applications of biotechnology. In order to achieve these advantages and control these multiphase flows, it is necessary to understand their generation and transport characteristics as influenced by geometrical miniaturization, channel wall properties, the effects of surfactants and operating conditions. For gas-liquid two-phase flow, dry air and deionized water were driven into hot embossed PMMA microchannels with 200 μm square test microchannels. Flow regimes, flow maps and the lengths of the gas bubbles and liquid plugs in terms of the liquid volumetric flow ratio (βL) were determined. Continuous generation of regular segmented flow was also discussed. Three sub-regimes of the Segmented flow were identified based on the statistical phase length scales observed over a substantial test channel length. For the liquid-liquid segmented flow, deionized water and perfluorocarbon with a surfactant were used as test fluids in the hot embossed polycarbonate microchannels. The effects of three expansion ratios from the injection to the test channels of 2, 4, and 16 were investigated comparing the flow regimes, transitions and maps in terms of a fixed carrier fluid volumetric flow ratio. The length of the dispersed fluids and the distance between consecutive droplets or plugs in terms of the carrier fluid volumetric flow ratio (βC) were determined. Velocities of the dispersed droplets and plugs were measured using double-pulsed laser illumination and were found to be 1.46 ± 0.08 and 1.25 ± 0.05 times faster than the superficial velocity of the segmented flow, respectively. The multiphase flow pressure drops were measured for all of the flow regimes in gas-liquid two-phase and liquid-liquid segmented flows. Each flow regime identified on the basis of topological observations, including the length scale of each fluid phase and the number of the gas bubbles or dispersed droplets in unit length with respect to the volumetric flow ratio, was associated with different trends in the pressure drop variation

Capillary and microchip gel electrophoresis using multiplexed fluorescence detection with both time-resolved and spectral-discrimination capabilities: applications in DNA sequencing using near-infrared fluorescence

Increasing the information content obtainable from a single assay and system miniaturization has continued to be important research areas in analytical chemistry. The research presented in this dissertation involves the development of a two-color, time-resolved fluorescence microscope for the acquisition of both steady-state and time-resolved data during capillary and microchip electrophoresis. The utility of this hybrid fluorescence detector has been demonstrated by applying it to DNA sequencing applications. Coupling color discrimination with time-resolved fluorescence offers increased multiplexing capabilities because the lifetime data adds another layer of information. An optical fiber-based fluorescence microscope was constructed, which utilized fluorescence in near-IR region, greatly simplifying the hardware and allowing superior system sensitivity. Time-resolved data was processed using electronics configured in a time-correlated single photon counting format. Cross-talk between color channels was successfully eliminated by utilizing the intrinsic time-resolved capability associated with the detector. The two-color, time-resolved microscope was first coupled to a single capillary and carried out two-color, two-lifetime sequencing of an M13 template, achieving a read length of 650 bps at a calling accuracy of 95.1%. The feasibility of using this microscope with microchips (glass-based chips) for sequencing was then demonstrated. Results from capillaries and microchips were compared, with the microchips providing faster analysis and adequate electrophoretic performance. Lifetimes of a set of fluorescent dyes were determined with favorable precision, in spite of the low loading levels associated with the microchips. The sequencing products were required to be purified and concentrated prior to electrophoretic sorting to improve data quality. PMMA-based microchips for DNA sequencing application were evaluated. The microchips were produced from thermo plastics, which allowed rapid and inexpensive production of microstructures with high aspect ratios. It was concluded that surface coating was needed on the polymer chips in order to achieve single-base resolution required for DNA sequencing. The capability of the two-color time-resolved microscope operated in a scanning mode was further explored. The successful construction of the scanner allows scanning of multi-channel microchips for high throughput processing

Production of uniform droplets using membrane, microchannel and microfluidic emulsification devices

This review provides an overview of major microengineering emulsification techniques for production of monodispersed droplets. The main emphasis has been put on membrane emulsification using Shirasu Porous Glass and microsieve membrane, microchannel emulsification using grooved-type and straight-through microchannel plates, microfluidic junctions and flow focusing microfluidic devices. Microfabrication methods for production of planar and 3D poly(dimethylsiloxane) devices, glass capillary microfluidic devices and single-crystal silicon microchannel array devices have been described including soft lithography, glass capillary pulling and microforging, hot embossing, anisotropic wet etching and deep reactive ion etching. In addition, fabrication methods for SPG and microseive membranes have been outlined, such as spinodal decomposition, reactive ion etching and ultraviolet LIGA (Lithography, Electroplating, and Moulding) process. The most widespread application of micromachined emulsification devices is in the synthesis of monodispersed particles and vesicles, such as polymeric particles, microgels, solid lipid particles, Janus particles, and functional vesicles (liposomes, polymersomes and colloidosomes). Glass capillary microfluidic devices are very suitable for production of core/shell drops of controllable shell thickness and multiple emulsions containing a controlled number of inner droplets and/or inner droplets of two or more distinct phases. Microchannel emulsification is a very promising technique for production of monodispersed droplets with droplet throughputs of up to 100 l h−1

Integrating Micro-Scale Separations to Matrix Assisted Laser Desorption and Ioniation Time of Flight Mass Spectrometry (MALDI-TOF-MS) for Protein Analysis

This dissertation describes the integration of micro-scale separations to matrix assisted laser desorption and ionization time-of-flight mass spectrometry (MALDI TOF MS) for protein analysis. MALDI MS provides unsurpassed accurate mass measurements of intact bio-molecules, for example peptides and proteins, which in turn generate high molecular specificity enabling the identity, function and structure of these molecules to be characterized. However, in order to realize the full potential of MS in proteomic studies, integrated sample processing on automated and high throughput platforms is required to address the complexity, diversity and the dynamic range of proteomic analysis. The work described here contributes towards the development of automated and high throughput micro-total analysis systems (µ-TAS) for proteomics. An overview of mass spectrometry instrumentation and techniques used in protein analysis is presented to highlight the significance of the work described. Microfluidics devices can serve as automated and high throughput platforms for integrating proteomics sample processing steps such as whole cell lyses, enrichment, solubilization, denaturation, protein separations, proteolytic digestion and chromatographic separations of peptides prior to MALDI TOF MS analysis. Therefore, coupling microfluidics devices to biological mass spectrometry is the first logical step towards developing fully integrated and automated systems for protein analysis. On-line and off-line approaches for analysis from microfluidic devices are discussed. The development of a specially tailored rotating ball inlet for automated on-line MALDI MS sample introduction from an electrophoresis-based separation platform is described. Electrophoresis-based micro-scale separations of peptides on fused silica capillary and polymer-based microfluidic devices were coupled to on-line MALDI TOF MS using a rotating ball inlet. The rotating ball inlet allowed for individual technique optimization and automation thereby eliminating the need for fractionation and routine MALDI sample preparation. High throughput solid phase micro-reactors for efficient enzymatic cleavages and improved protein identification with MALDI MS in a microfluidic device were also developed for incorporation in an integrated protein analysis microfluidic system. Future work that outlines the framework and focus geared towards integrating the modules discussed in this dissertation into a functional micro-total analysis system for protein sample processing is discussed