For the first time we demonstrate a self-sufficient lab-on-a-foil system for the fully automated analysis of nucleic acids which is based on the recently available isothermal recombinase polymerase amplification (RPA). The system consists of a novel, foil-based centrifugal microfluidic cartridge including prestored liquid and dry reagents, and a commercially available centrifugal analyzer for incubation at 37 degrees C and real-time fluorescence detection. The system was characterized with an assay for the detection of the antibiotic resistance gene mecA of Staphylococcus aureus. The limit of detection was <10 copies and time-to-result was <20 min. Microfluidic unit operations comprise storage and release of liquid reagents, reconstitution of lyophilized reagents, aliquoting the sample into < or = 30 independent reaction cavities, and mixing of reagents with the DNA samples. The foil-based cartridge was produced by blow-molding and sealed with a self-adhesive tape. The demonstrated system excels existing PCR based lab-on-a-chip platforms in terms of energy efficiency and time-to-result. Applications are suggested in the field of mobile point-of-care analysis, B-detection, or in combination with continuous monitoring systems.
This critical review is motivated by an increasing interest of the microfluidics community in developing complete Lab-on-a-Chip solutions based on thin and flexible films (Lab-on-a-Foil). Those implementations benefit from a broad range of fabrication methods that are partly adopted from well-established macroscale processes or are completely new and promising. In addition, thin and flexible foils enable various features like low thermal resistance for efficient thermocycling or integration of easily deformable chambers paving the way for new means of on-chip reagent storage or fluid transport. From an economical perspective, Lab-on-a-Foil systems are characterised by low material consumption and often low-cost materials which are attractive for cost-effective high-volume fabrication of self-contained disposable chips. The first part of this review focuses on available materials, fabrication processes and approaches for integration of microfluidic functions including liquid control and transport as well as storage and release of reagents. In the second part, an analysis of the state of Lab-on-a-Foil applications is provided with a special focus on nucleic acid analysis, immunoassays, cell-based assays and home care testing. We conclude that the Lab-on-a-Foil approach is very versatile and significantly expands the toolbox for the development of Lab-on-a-Chip solutions.
We present a novel, cost-efficient process chain for fast tooling and small-lot replication of high-quality, multi-scale microfluidic polymer chips within less than 5 days. The fabrication chain starts with a primary master which is made by well-established cleanroom processes such as DRIE or negative SU-8 resist based surface micromachining. The formation of undercuts in the master which would complicate demolding is carefully avoided. Secondary PDMS masters or epoxy-based masters which are more suitable for common polymer replication schemes such as soft-embossing, hot-embossing or injection molding are subsequently cast from the primary masters. The polymer replica are mainly made of COC and show excellent fidelity with the conventionally micromachined master while displaying no degeneration, even after more than 200 cycles. The use of other polymers such as PMMA is also possible. The process chain further includes surface modification techniques for overall, long-term stable hydrophilic coatings and for local hydrophobic patches as well as a durable sealing based on thermal bonding.
We present a novel process flow enabling prototyping of microfluidic cartridges made out of polymer films. Its high performance is proven by implementation of a microfluidic genotyping assay testing 22 DNA samples including clinical isolates from patients infected by methicilin-resistant Staphylococcus aureus (MRSA). The microfluidic cartridges (disks) are fabricated by a novel process called microthermoforming by soft lithography (microTSL). Positive moulds are applied allowing for higher moulding precision and very easy demoulding when compared to conventional microthermoforming. High replication accuracies with geometric disk-to-disk variations of less than 1% are typical. We describe and characterise fabrication and application of microfluidic cartridges with wall thicknesses <188 microm thus enabling efficient thermocycling during real-time polymerase chain reaction (PCR). The microfluidic cartridges are designed for operation in a slightly modified commercial thermocycling instrument. This approach demonstrates new opportunities for both microfluidic developments and well-established laboratory instruments. The microfluidic protocol is controlled by centrifugal forces and divides the liquid sample parallely into independent aliquots of 9.8 microl (CV 3.4%, N = 32 wells). The genotyping assays are performed with pre-stored primers and probes for real-time PCR showing a limit of detection well below 10 copies of DNA per reaction well (N = 24 wells in 3 independent disks). The system was evaluated by 44 genotyping assays comprising 22 DNA samples plus duplicates in a total of 11 disks. The samples contained clinical samples of seven different genotypes of MRSA as well as positive and negative controls. The results are in excellent agreement with the reference in microtubes.
Biofilm-associated infections of medical devices are a global problem. For the prevention of such infections, biomaterial surfaces are chemically or topographically modified to slow down the initial stages of biofilm formation. In the bifunctional material here presented, chemical and topographical cues are combined, so that protein and bacterial adhesion as well as bacterial proliferation are effectively inhibited. Upon changes in the surface topography parameters and investigation of the effect of these changes on bioactivity, structure−property relationships are obtained. The target material is obtained by microcontact printing (μCP), a soft lithography method. The antimicrobial component, poly(oxanorbornene)-based synthetic mimics of an antimicrobial peptide (SMAMP), was printed onto a protein-repellent polysulfobetaine hydrogel, so that bifunctional 3D structured polymer surfaces with 1, 2, and 8.5 μm spacing are obtained. These surfaces are characterized with fluorescence microscopy, surface plasmon resonance spectroscopy, atomic force microscopy, and contact angle measurements. Biological studies show that the bifunctional surfaces with 1 and 2 μm spacing are 100% antimicrobially active against Escherichia coli and Staphylococcus aureus, 100% fibrinogen-repellent, and nontoxic to human gingival mucosal keratinocytes. At 8.5 μm spacing, the broad-band antimicrobial activity and the protein repellency are compromised, which indicates that this spacing is above the upper limit for effective simultaneous antimicrobial activity and protein repellency of polyzwitterionic−polycationic materials.
In this paper we introduce the three-dimensional formulation of the OPTOS formalism, a matrix-based method that allows for the efficient simulation of non-coherent light propagation and absorption in thick textured sheets. As application examples, we calculate the absorptance of solar cells featuring textures on front and rear side with different feature sizes operating in different optical regimes. A discretization of polar and azimuth angle enables a three-dimensional description of systems with arbitrary surface textures. We present redistribution matrices for 3D surface textures, including pyramidal textures, binary crossed gratings and a Lambertian scatterer. The results of the OPTOS simulations for silicon sheets with different combinations of these surfaces are in accordance with both optical measurements and results based on established simulation methods like ray tracing. Using OPTOS, we show that the integration of a diffractive grating at the rear side of a silicon solar cell featuring a pyramidal front side results in absorption close to the Yablonovitch Limit enhancing the photocurrent density by 0.6 mA/cm2 for a 200 µm thick cell.
We demonstrate the use of photosensitive epoxy laminate TMMF S2045 for the fabrication and sealing of tapered microfluidic channels. The 45 μm thick resist enables the fabrication of shallow sealed cavities featuring extreme aspect ratios of less than 1:40 (h = 45 μm, w = 2000 μm). It also provides high resolution and enables minimum feature sizes of 10 μm. For the fabrication of free-standing structures, an aspect ratio of up to 7:1 was achieved. The dry-film photoresist can be applied easily by lamination onto structured substrates. The total thickness variation of the resist across a 100 mm wafer was determined to be less than ±0.6 μm. Process parameters for the fabrication and sealing of various micro-channels are discussed and optimized in this paper. The main focus was to minimize thermal impact during lamination, soft-bake, exposure and post-exposure bake, which could lead to lid sagging or channel clogging due to liquefaction of uncured resist. We tested TMMF according to ISO 10995-5 and found it to be non-cytotoxic, enabling its use for biological applications. Swelling of less than 5% for incubation of the dry-film resist in several biologically relevant solvents, buffers and cleaning solutions was observed.
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