Proyag Datta

Simple replication methods for producing nanoslits in thermoplastics and the transport dynamics of double-stranded DNA through these slits

Mixed-scale nano- and microfluidic networks were fabricated in thermoplastics using simple and robust methods that did not require the use of sophisticated equipment to produce the nanostructures. High-precision micromilling (HPMM) and photolithography were used to generate mixed-scale molding tools that were subsequently used for producing fluidic networks into thermoplastics such as poly(methyl methacrylate), PMMA, cyclic olefin copolymer, COC, and polycarbonate, PC. Nanoslit arrays were imprinted into the polymer using a nanoimprinting tool, which was composed of an optical mask with patterns that were 2-7 µm in width and a depth defined by the Cr layer (100 nm), which was deposited onto glass. The device also contained a microchannel network that was hot embossed into the polymer substrate using a metal molding tool prepared via HPMM. The mixed-scale device could also be used as a master to produce a polymer stamp, which was made from polydimethylsiloxane, PDMS, and used to generate the mixed-scale fluidic network in a single step. Thermal fusion bonding of the cover plate to the substrate at a temperature below their respective T(g) was accomplished by oxygen plasma treatment of both the substrate and cover plate, which significantly reduced thermally induced structural deformation during assembly: ∼6% for PMMA and ∼9% for COC nanoslits. The electrokinetic transport properties of double-stranded DNA (dsDNA) through the polymeric nanoslits (PMMA and COC) were carried out. In these polymer devices, the dsDNA demonstrated a field-dependent electrophoretic mobility with intermittent transport dynamics. DNA mobilities were found to be 8.2 ± 0.7 × 10(-4) cm(2) V(-1) s(-1) and 7.6 ± 0.6 × 10(-4) cm(2) V(-1) s(-1) for PMMA and COC, respectively, at a field strength of 25 V cm(-1). The extension factors for λ-DNA were 0.46 in PMMA and 0.53 in COC for the nanoslits (2-6% standard deviation).

Fabrication of a cyclic olefin copolymer planar waveguide embedded in a multi-channel poly(methyl methacrylate) fluidic chip for evanescence excitation

The fabrication and characterization of a novel cyclic olefin copolymer (COC) waveguide embedded in a poly(methyl methacrylate), PMMA, fluidic chip configured in a multi-channel format with an integrated monolithic prism for evanescent fluorescence excitation are reported. The fabrication approach allowed the embedded waveguide to be situated orthogonal to a series of fluidic channels within the PMMA wafer to sample fluorescent solutions in these channels using the evanescence properties of the waveguide. Construction of the device was achieved using several fabrication techniques including high precision micromilling, hot embossing and stenciling of a polymer melt to form the waveguide and coupling prism. A waveguide channel was fabricated in the fluidic chips cover plate, also made from PMMA, and was loaded with a COC solution using a pre-cast poly(dimethylsiloxane), PDMS, stencil containing a prism-shaped recess. The PMMA substrate contained multiple channels (100 microm wide x 30 microm deep with a pitch of 100 microm) that were situated orthogonal to the waveguide to allow penetration of the evanescent field into the sampling solution. The optical properties of the waveguide in terms of its transmission properties and penetration depth of the evanescent field in the adjacent solution were evaluated. Finally, the device was used for laser-induced fluorescence evanescent excitation of a dye solution hydrodynamically flowing through multiple microfluidic channels in the chip and processed using a microscope equipped with a charge-coupled device (CCD) for parallel readout. The device and optical system were able to image 11 channels simultaneously with a limit-of-detection of 7.1 x 10(-20) mol at a signal-to-noise ratio of 2. The waveguide was simple to manufacture and could be scaled to illuminate much higher channel numbers making it appropriate for high-throughput measurements using evanescent excitation.

Fully Integrated Thermoplastic Genosensor for the Highly Sensitive Detection and Identification of Multi‐Drug‐Resistant Tuberculosis

Infectious diseases are a major global health burden accounting for approximately 15 million deaths annually, many from drug resistant pathogenic agents, with a significant number of cases occurring in developing countries.[1–7] In particular, the resurgence of tuberculosis (TB) has been accompanied by the rapid spread of multi-drug resistance TB (MDR-TB) resulting from Mycobacterium tuberculosis (Mtb) strains that fail to respond to the first-line drugs, rifampin and isoniazid. Currently, <5% of ~0.5 million MDR-TB cases estimated globally are appropriately diagnosed and treated due in part to the long assay turnaround time associated with conventional culture-based drug susceptibility testing.[8]

Titer plate formatted continuous flow thermal reactors for high throughput applications: fabrication and testing

A high throughput, multi-well (96) polymerase chain reaction (PCR) platform, based on a continuous flow (CF) mode of operation, was developed. Each CFPCR device was confined to a footprint of 8 × 8 mm2, matching the footprint of a well on a standard micro-titer plate. While several CFPCR devices have been demonstrated, this is the first example of a high-throughput multi-well continuous flow thermal reactor configuration. Verification of the feasibility of the multi-well CFPCR device was carried out at each stage of development from manufacturing to demonstrating sample amplification. The multi-well CFPCR devices were fabricated by micro-replication in polymers, polycarbonate to accommodate the peak temperatures during thermal cycling in this case, using double-sided hot embossing. One side of the substrate contained the thermal reactors and the opposite side was patterned with structures to enhance thermal isolation of the closely packed constant temperature zones. A 99 bp target from a λ-DNA template was successfully amplified in a prototype multi-well CFPCR device with a total reaction time as low as ~5 min at a flow velocity of 3 mm s−1 (15.3 s cycle−1) and a relatively low amplification efficiency compared to a bench-top thermal cycler for a 20-cycle device; reducing the flow velocity to 1 mm s−1 (46.2 s cycle−1) gave a seven-fold improvement in amplification efficiency. Amplification efficiencies increased at all flow velocities for 25-cycle devices with the same configuration.

Development of an Integrated Polymer Microfluidic Stack

Microfluidic is a field of considerable interest. While significant research has been carried out to develop microfluidic components, very little has been done to integrate the components into a complete working system. We present a flexible modular system platform that addresses the requirements of a complete microfluidic system. A microfluidic stack system is demonstrated with the layers of the stack being modular for specific functions. The stack and accompanying infrastructure provides an attractive platform for users to transition their design concepts into a working microfluidic system quickly with very little effort. The concept is demonstrated by using the system to carry out a chemilumiscence experiment. Details regarding the fabrication, assembly and experimental methods are presented.

LiGA Research and Service at CAMD

Since 1995 CAMD has been offering exposure services, so called print shop for a variety of users interested in making precision High-Aspect-Ratio Microstructures (HARMST) for various application. Services have been expanded beyond only the print shop service in recent years and now include x-ray mask fabrication, substrate preparation for PMMA and SU- 8 resists, electroplating, finishing and molding. Metallic and polymeric parts are now routinely fabricated for precision engineering, micro-fluidic and micro-optic applications. This paper presents a brief overview of the actual status of LiGA services provided at CAMD including ongoing research efforts and examples of LiGA components for different applications.

Passive alignment structures in modular, polymer microfluidic devices

For connecting polymeric, modular microfluidic devices, precise, passive alignment structures can prevent infinitesimal motions between the devices and minimize misalignment of the devices. The motion and constraint of passive alignment structures were analyzed for the design of assembly features using screw theory. A combination of three v-groove and sphere joints constrained all degrees of freedom of the two mating plates without over-constraint. To validate the designed passive alignment scheme, hot embossing experiments were conducted using a micromilled brass mold insert, containing alignment features. Prototype alignment structures have dimensional variation. The alignment accuracy of the stacked polymeric plates was estimated by the mismatches between alignment marks of two plates. The mismatches ranged from 28 μm to 70 μm.Copyright

Optimization of Geometry for Continuous Flow PCR Devices in a Titer Plate-Based PCR Multi-Reactor Platform

A highly parallel, polymerase chain reaction (PCR) multireactor platform is in high demand to satisfy the high throughput requirements for exploiting the accumulated genetic information from the Human Genome Project. By incorporating continuous flow PCR (CFPCR) devices in a polymer 96-well titer plate format, DNA amplification can be performed with steady-state temperature control and faster reaction speed at lower cost. Prior to the realization of a PCR multi-reactor platform, consisting of a sample delivery chip, a PCR multireactor chip, and a thermal cycler, optimization of the geometry for CFPCR devices in a titer plate-based PCR multi-reactor chip based on manufacturing feasibility is necessary. A prototype PCR multi-reactor chip was designed in a 96-well titer plate format with twelve different CFPCR configurations. High quality metallic, large area mold inserts (LAMIs) were fabricated using an SU-8 based UV-LIGA technique by overplating nickel in SU-8 electroplating templates. Micro molding of polycarbonate (PC) was done using hot embossing, resulting in good replication fidelity over the large surface area. Thermal fusion bonding of the molded PC chips using a custom-made bonding jig yielded acceptable sealing results. The manufacturability investigation throughout the design and the process sequence suggested that the microchannel walls require a minimum width of at least 20 μm and an aspect ratio of 2 for structural rigidity. An optimal CFPCR device for use in a PCR multi-reactor chip can be selected with a series of amplification experiments with the development of a thermal cycler.© 2007 ASME

Polymeric waveguides for orthogonal near surface fluorescent excitation

The objective of this effort was to fabricate a waveguide integrated in a polymer microfluidic chip in order to deliver excitation light to fluorescent probes contained in a fluidic channel. Instead of exciting the volume at a certain point along a fluidic channel, the goal herein was to excite all the probes contained along the length of the fluidic channel. An air-waveguide structure was designed and integrated into a polymer microfluidic chip. Fabrication of the microfluidic chip was carried out by double-sided hot embossing of poly methyl methacrylate (PMMA) in sheet form. The efficacy of the waveguide was evaluated by coupling light from a laser into it and testing the fluorescence intensity from dye contained in the microfluidic channel. The results demonstrate illumination of the entire length of the microfluidic channel with excitation wavelength light from the waveguide. Details of the design, fabrication process and initial experimental results are presented in the course of this paper.

Microfluidic package design for magnetoresistive biosensors

In order for Magnetoresistive Biosensor technology to become a mainstream product for clinical and consumer use, several outstanding technical issues must be solved. This paper will focus on one of those issues, which is the need to adapt standard semiconductor packaging processes to fall within some biosensor fabrication process constraints. A set of materials and interconnection methods that meet these biosensor requirements are presented. The resulting architecture is compatible with laboratory assembly, but can be scaled up to small and medium manufacturing quantities by using larger 2-dimensional areas per production batch.