Roll-to-roll UV nanoimprint lithography has superior advantages for high-throughput manufacturing of micro- or nano-structures on flexible polymer foils with various geometries and configurations. Our pilot line provides large-scale structure imprinting for cost-effective polymer biochips (4500 biochips/hour), enabling rapid and multiplexed detections. A complete high-volume process chain of the technology for producing structures like μ-sized, triangular optical out-couplers or capillary channels (width: from 1 μm to 2 mm, height: from 200 nm up to 100 μm) to obtain biochips (width: 25 mm, length: 75 mm, height: 100 μm to 1.5 mm) was described. The imprinting process was performed with custom-developed resins on polymer foils with resin thicknesses ranging between 125-190 μm. The produced chips were tested in a commercial point-of-care diagnostic system for multiplexed DNA analysis of methicillin resistant Staphylococcus aureus (e.g., mecA, mecC gene detections). Specific target DNA capturing was based on hybridisation between surface bound DNA probes and biotinylated targets from the sample. The immobilised biotinylated targets subsequently bind streptavidin-horseradish peroxidase conjugates, which in turn generate light upon incubation with a chemiluminescent substrate. To enhance the light out-coupling thus to improve the system performance, optical structures were integrated into the design. The limits-of-detection of mecA (25 bp) for chips with and without structures were calculated as 0.06 and 0.07 μM, respectively. Further, foil-based chips with fluidic channels were DNA functionalised in our roll-to-roll micro-array spotter following the imprinting. This straightforward approach of sequential imprinting and multiplexed DNA functionalisation on a single foil was also realised for the first time. The corresponding foil-based chips were able to detect mecA gene DNA sequences down to a 0.25 μM concentration.

Reliable bonding of microstructured polymer parts is one of the major challenges in industrial fabrication of microfluidic devices. In the present work, the effects of a UV/ozone surface activation on the bonding process were investigated for the combination of a commonly used thermoplastic cyclic olefin copolymer (COC) with an elastomeric COC (eCOC) as a new thermoplastic elastomer material. Bonding was studied using two-component injection molded parts of COC and eCOC, together with microfluidic COC chips. Surface activation and bonding process parameters were optimized and bond strengths were characterized by the wedge test method. The results showed that strong bonding of this polymer materials combination can be achieved at temperatures significantly below the bulk glass transition temperature of COC.

A multiparameter gas sensor based on distributed feedback interband cascade lasers emitting at 4.35 μm and ultrafast electro-spun luminescence oxygen sensors has been developed for the quantification and continuous monitoring of 13CO2/12CO2 isotopic ratio changes and oxygen in exhaled mouse breath samples. Mid-infrared absorption spectra for quantitatively monitoring the enrichment of 13CO2 levels were recorded in a miniaturized dual-channel substrate-integrated hollow waveguide using balanced ratiometric detection, whereas luminescence quenching was used for synchronously detecting exhaled oxygen levels. Allan variance analysis verified a CO2 measurement precision of 1.6‰ during a 480 s integration time. Routine online monitoring of exhaled mouse breath was performed in 14 mechanically ventilated and instrumented mice and demonstrated the feasibility of online isotope-selective exhaled breath analysis within microliters of probed gas samples using the reported combined sensor platform.

Highly fluorescent and photostable (2-alkyl)-1H-benzo[de]isoquinoline-1,3(2H)-diones with a polymerizable norbornene scaffold have been synthesized and polymerized using ring-opening metathesis polymerization. The monomers presented herein could be polymerized in a living fashion, using different comonomers and different monomer ratios. All obtained materials showed good film-forming properties and bright fluorescence caused by the incorporated push–pull chromophores. Additionally, one of the monomers containing a methylpiperazine functionality showed protonation-dependent photoinduced electron transfer which opens up interesting applications for logic gates and sensing.

The equipment of cellulose ultrathin films with BSA (bovine serum albumin) via cationization of the surface by tailor-made cationic celluloses is described. In this way, matrices for controlled protein deposition are created, whereas the extent of protein affinity to these surfaces is controlled by the charge density and solubility of the tailored cationic cellulose derivative. In order to understand the impact of the cationic cellulose derivatives on the protein affinity, their interaction capacity with fluorescently labeled BSA is investigated at different concentrations and pH values. The amount of deposited material is quantified using QCM-D (quartz crystal microbalance with dissipation monitoring, wet mass) and MP-SPR (multi-parameter surface plasmon resonance, dry mass), and the mass of coupled water is evaluated by combination of QCM-D and SPR data. It turns out that adsorption can be tuned over a wide range (0.6-3.9 mg dry mass m(-2)) depending on the used conditions for adsorption and the type of employed cationic cellulose. After evaluation of protein adsorption, patterned cellulose thin films have been prepared and the cationic celluloses were adsorbed in a similar fashion as in the QCM-D and SPR experiments. Onto these cationic surfaces, fluorescently labeled BSA in different concentrations is deposited by an automatized spotting apparatus and a correlation between the amount of the deposited protein and the fluorescence intensity is established.

Simultaneous antibacterial and anticoagulant surfaces have been prepared by immobilization of engineered gold nanoparticles onto different kinds of surfaces. The gold nanoparticle core is surrounded by a hemocompatible, anticoagulant polysaccharide, 6-O chitosan sulfate, which serves as reduction and stabilizing agent for the generation of gold nanoparticles in a microwave mediated reaction. The particle suspension shows anticoagulant activity, which is investigated by aPTT and PT testing on citrated blood samples of three patients suffering from congenital or acquired bleeding disorders. The amount of nanoparticles deposited on the surfaces is quantified by a quartz crystal microbalance with dissipation unit. All gold containing surfaces exhibit excellent antimicrobial properties against the chosen model organism, Escherichia coli MG 1655 [R1-16]. Moreover, blood plasma coagulation times of the surfaces are increased after deposition of the engineered nanoparticles as demonstrated by QCM-D.

Patterned surface functionalization of PDMS with the biopolymer cellulose via lithographic methods.

For most “simple” analytes (e.g. oxygen), the actual sensing element of optochemical sensors consists of a luminescent dye embedded in a polymeric matrix whose luminescence intensity and decay time correlates with the analyte concentration. The response characteristics of these sensors are limited by the time requirement for diffusion and equilibration of the analyte in the bulk of the matrix material. Traditionally, the sensor formulation is cast onto a substrate as a compact layer. Thin layers result in good response speed but poor signal intensity and signal-to-noise ratio, respectively. By means of electrospinning, many polymers can be processed to nanofibers, resulting in a highly-porous textile-like layer with high surface-to-volume ratio and superb analyte accessibility. Doped with fluorescent dyes, such fiber layers are well suited for fast sensing applications where the response time is a critical issue. It was shown that by electrospinning of a typical oxygen sensor formulation (PtTFPP immobilized in polystyrene), that the response time t90 can be accelerated from several seconds to less than 40ms at comparable general sensor performance. This work focuses on the long-term operational stability of such nanofiber based optical sensors. No pronounced tendency for degradation could be observed in long-term stress tests.

Two different surface activation methods (UV/ozone and oxygen plasma treatment) were applied for patterned surface activation of cyclo-olefin polymer (COP) surfaces combined with different masking techniques (metal shadow mask and protective tape). Surface properties were characterized by various methods such as contact angle measurement, ATR-IR, XPS and Surface enhanced ellipsometric contrast (SEEC) microscopy. UV/ozone and oxygen plasma allowed for patterned surface modification of COP leading to the formation of carboxylic and hydroxyl groups on the activated part of the surface. Stability against organic solvents was determined by rinsing the activated substrates with 2-propanol. For UV/ozone treatment it was found that a thin film of degradation products remains on the COP surface and is at least partly removed in the following washing or rinsing steps.

The use of organic photodiodes (OPDs) for measuring phosphorescent lifetimes of optochemical oxygen sensors is described. Phosphorescent indicators with lifetimes ranging from ∼5 to 60 μs have been studied using light-emitting diodes as the excitation source and organic photodiodes integrated into the sensor substrate for detection. A measurement system using an adjusted electronic circuitry to detect photocurrents in the nanoampere range is presented. The response behaviour of the organic photodiodes has been characterized, and it was found that a forward (positive) bias had to be applied in order to decrease the response time of the OPDs to a range suitable for phosphorescence decay time measurements. A modulation cutoff frequency of ∼100 kHz has been determined, corresponding to a response time of the organic photodiodes of 1.6 μs. Two sensor dyes have been characterized regarding their lifetimes upon exposure to 0–20 % oxygen, and it was shown that results comparable to literature data and inorganic photodetectors can be achieved.

Breath analysis is an attractive non-invasive strategy for early disease recognition or diagnosis, and for therapeutic progression monitoring, as quantitative compositional analysis of breath can be related to biomarker panels provided by a specific physiological condition invoked by e.g., pulmonary diseases, lung cancer, breast cancer, and others. As exhaled breath contains comprehensive information on e.g., the metabolic state, and since in particular volatile organic constituents (VOCs) in exhaled breath may be indicative of certain disease states, analytical techniques for advanced breath diagnostics should be capable of sufficient molecular discrimination and quantification of constituents at ppm-ppb - or even lower - concentration levels.While individual analytical techniques such as e. g., mid-infrared spectroscopy may provide access to a range of relevant molecules, some IR-inactive constituents require the combination of IR sensing schemes with orthogonal analytical tools for extended molecular coverage. Combining mid-infrared hollow waveguides (HWGs) with luminescence sensors (LS) appears particularly attractive, as these complementary analytical techniques allow to simultaneously analyze total CO2 (via luminescence), the (CO2)-C-12/(CO2)-C-13 tracer-to-tracee (TTR) ratio (via IR), selected VOCs (via IR) and O-2 (via luminescence) in exhaled breath, yet, establishing a single diagnostic platform as both sensors simultaneously interact with the same breath sample volume.In the present study, we take advantage of a particularly compact (shoebox-size) FTIR spectrometer combined with novel substrate-integrated hollow waveguide (iHWG) recently developed by our research team, and miniaturized fiberoptic luminescence sensors for establishing a multi-constituent breath analysis tool that is ideally compatible with mouse intensive care stations (MICU). Given the low tidal volume and flow of exhaled mouse breath, the TTR is usually determined after sample collection via gas chromatography coupled to mass spectrometric detection. Here, we aim at potentially continuously analyzing the TTR via iHWGs and LS flow-through sensors requiring only minute (< 1 mL) sample volumes. Furthermore, this study explores non-linearities observed for the calibration functions of (CO2)-C-12 and (CO2)-C-13 potentially resulting from effects related to optical collision diameters e.g., in presence of molecular oxygen. It is anticipated that the simultaneous continuous analysis of oxygen via LS will facilitate the correction of these effects after inclusion within appropriate multivariate calibration models, thus providing more reliable and robust calibration schemes for continuously monitoring relevant breath constituents.

We demonstrate screen printing of polymer waveguides on rigid and flexible substrates. The waveguides are characterized by optical microscopy and propagation losses are determined by cut-back measurements. Furthermore, the efficiency of light coupling using fluorescent molecules within the waveguides is measured with regard to the application feasibility in sensor systems. It is shown that minimum propagation losses of 0.74 dB/cm can be reached with this low-cost printing technique.

A novel optical sensor device monolithically integrated on a glass capillary is presented. Therefore, we took advantage of the ability to fabricate organic optoelectronic devices on non-planar substrates. The functionality of the concept is demonstrated by realizing an integrated oxygen sensor based on luminescence decay time measurement.

There is a steadily growing demand for miniaturized bioanalytical devices allowing for on-site or point-of-care detection of biomolecules or pathogens in applications like diagnostics, food testing, or environmental monitoring. These, so called labs-on-a-chip or micro-total analysis systems (mu-TAS) should ideally enable convenient sample-in - result-out type operation. Therefore, the entire process from sample preparation, metering, reagent incubation, etc. to detection should be performed on a single disposable device (on-chip). In the early days such devices were mainly fabricated using glass or silicon substrates and adapting established fabrication technologies from the electronics and semiconductor industry. More recently, the development focuses on the use of thermoplastic polymers as they allow for low-cost high volume fabrication of disposables. One of the most promising materials for the development of plastic based lab-on-a-chip systems are cyclic olefin polymers and copolymers (COP/COC) due to their excellent optical properties (high transparency and low autofluorescence) and ease of processing.We present a bioanalytical system for whole blood samples comprising a disposable plastic chip based on TIRF (total internal reflection fluorescence) optical detection. The chips were fabricated by compression moulding of COP and microfluidic channels were structured by hot embossing. These microfluidic structures integrate several sample pretreatment steps. These are the separation of erythrocytes, metering of sample volume using passive valves, and reagent incubation for competitive bioassays. The surface of the following optical detection zone is functionalized with specific capture probes in an array format. The plastic chips comprise dedicated structures for simple and effective coupling of excitation light from low-cost laser diodes. This enables TIRF excitation of fluorescently labeled probes selectively bound to detection spots at the microchannel surface. The fluorescence of these detection arrays is imaged using a simple set-up based on a digital consumer camera. Image processing for spot detection and intensity calculation is accomplished using customized software. Using this combined TIRF excitation and imaging based detection approach allowes for effective suppression of background fluorescence from the sample, multiplexed detection in an array format, as well as internal calibration and background correction.

The use of cationic biopolymer surfaces for high protein binding affinity matrices is described. As model proteins, fluorescently labeled bovine serum albumins (FITC-BSA, TRITC-BSA) have been employed. The amount of proteins on such cationically rendered surfaces was quantified by QCM-D. In addition, flexible, transparent, patterned COP slides have been prepared and loaded with proteins ranging from 15 pM to 15 μM TRITC-BSA.

Two different lithographic techniques for the patterning of thin biopolymer films are developed. The first method is based on using a microstructured elastomeric mold for the structuring of thin films of regenerated cellulose. The thin films are manufactured by spin-coating of trimethylsilyl cellulose (TMSC) and subsequent regeneration. The microchannels formed by the mold and the cellulose film are filled with a cellulase solution by capillary action. In the areas exposed to the enzyme solution, the cellulose film is digested, whereas the area in contact with the mold is protected from the enzymatic activity. Optical thickness measurements, atomic force microscopy and fluorescent staining confirm a successful patterning of cellulose on several substrates by this method. The second method is based on the structured regeneration of thin TMSC films. TMSC surfaces are protected with metal masks and exposed to vapors of hydrochloric acid. These treatments result in hydrophilic cellulose structures surrounded by hydrophobic TMSC with differing physicochemical properties. Treatments of the obtained structures with cellulases allow the selective removal of pure cellulose, whereas a TMSC pattern remains on the surface. These TMSC can be regenerated back to pure cellulose by treatments with vapors of hydrochloric acid. The developed methods allow the effective fabrication of micropatterned biopolymer thin films suitable for further functionalization and application in, e.g., bioanalytical devices. This is shown by the immobilization and detection of single-stranded DNA on structured cellulose surfaces. Owing to the versatility of both patterning approaches the methods can be further extended to other combinations of substrates and enzymes.

In this contribution we describe a microfluidic chip combining plasma separation, sample metering, dissolution/incubation with reagents stored on-chip and optical detection. The system allows defining the incubation time and works under constant externally applied pressure using only passive valves for actuation. This allowed the realization of a bioanalytical device for whole blood samples comprising a disposable plastic chip using TIRF (total internal reflection fluorescence) based optical detection of biochemical binding events.

There is a growing demand for functional layers for the immobilization of (bio)molecules on different kinds of substrates in the field of biosensors, microarrays, and lab-on-a-chip development. These functional coatings should have the ability to specifically bind (bio)molecules with a high binding efficiency, while showing low unspecific binding during the following assay. In this paper we present rSbpA surface layer proteins (S-layer proteins) as a versatile immobilization layer for the development of DNA microarrays. S-layer proteins show the ability to reassemble into two-dimensional arrays on solid surfaces and their functional groups, such as carboxylic groups, are repeated with the periodicity of the lattice, allowing for immobilization of other (bio)molecules. Different fluorescently labeled amino functionalized DNA oligomers were covalently linked to the S-layer matrices to allow the characterization of DNA binding on S-layers. Hybridization and dissociation of DNA-oligomers were studied on S-layer coated slides, revealing low levels of unspecific adsorption of DNA on S-layer based immobilization matrices. In the following the principle was transferred to a DNA microarray design showing successful spotting and hybridization on whole microarray slides. Besides common laser scanning for fluorescence detection, S-layer based microarrays were evaluated with a compact, low cost platform for direct fluorescence imaging based on surface plasmon enhanced fluorescence excitation. It could be shown that S-layer protein layers are promising as immobilization matrices for the development of biosensors and microarrays.