Leena Hakalahti

Patterned Immobilization of Antibodies within Roll-to-Roll Hot Embossed Polymeric Microfluidic Channels

This paper describes a method for the patterned immobilization of capture antibodies into a microfluidic platform fabricated by roll-to-roll (R2R) hot embossing on poly (methyl methacrylate) (PMMA). Covalent attachment of antibodies was achieved by two sequential inkjet printing steps. First, a polyethyleneimine (PEI) layer was deposited onto oxygen plasma activated PMMA foil and further cross-linked with glutaraldehyde (GA) to provide an amine-reactive aldehyde surface (PEI-GA). This step was followed by a second deposition of antibody by overprinting on the PEI-GA patterned PMMA foil. The PEI polymer ink was first formulated to ensure stable drop formation in inkjet printing and the printed films were characterized using atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS). Anti-CRP antibody was patterned on PMMA foil by the developed method and bonded permanently with R2R hot embossed PMMA microchannels by solvent bonding lamination. The functionality of the immobilized antibody inside the microfluidic channel was evaluated by fluorescence-based sandwich immunoassay for detection of C-reactive protein (CRP). The antibody-antigen assay exhibited a good level of linearity over the range of 10 ng/ml to 500 ng/ml (R2 = 0.991) with a calculated detection limit of 5.2 ng/ml. The developed patterning method is straightforward, rapid and provides a versatile approach for creating multiple protein patterns in a single microfluidic channel for multiplexed immunoassays.

Highly sensitive biosensor based on UV-imprinted layered polymeric–inorganic composite waveguides

An evanescent field sensor utilizing layered polymeric-inorganic composite waveguide configuration was developed in this work. The composite waveguide structure consists of a UV-imprint patterned polymer inverted rib waveguide with a Ta2O5 thin film sputter-deposited on top of the low refractive index polymer layers. The results suggest that the polymer based sensor can achieve a detection limit of 3 × 10(-7) RIU for refractive index sensing and corresponding limit of about 100 fg/mm2 for molecular adsorption detection. Besides enhancing the sensitivity significantly, the inorganic coating on the polymer layer was found to block water absorption effectively into the waveguide resulting in a stabilized sensor operation. The ability to use the developed sensor in specific molecular detection was confirmed by investigating antibody - antigen binding reactions. The results of this work demonstrate that high performance sensing capability can be obtained with the developed composite waveguide sensor.

Polymeric dual-slab waveguide interferometer for biochemical sensing applications.

A polymer based dual-slab waveguide Youngs interferometer was demonstrated for biochemical sensing. Evanescent field is utilized for probing the binding events of biomolecules on the waveguide surface. Refractive index sensing in analyte and protein adsorption on the sensing surface were investigated with glucose de-ionized water solution and bovine serum albumin, immunoglobulin G solutions in phosphate buffered saline buffer. A detection limit of 10(-5) RIU and 4 pg/mm(2) was achieved for homogeneous and surface sensing, respectively. Also, the influence of water absorption inside the polymeric device on the measurement stability was evaluated. The results indicate that the waveguide polymer sensor fabricated with the spin coating technique can achieve a satisfactory sensitivity for homogeneous refractive index sensing and, as well, for monitoring molecular binding events on the surface.

Versatile bio-ink for covalent immobilization of chimeric avidin on sol-gel substrates.

A bio-ink for covalent deposition of thermostable, high affinity biotin-binding chimeric avidin onto sol-gel substrates was developed. The bio-ink was prepared from heterobifunctional crosslinker 6-maleimidohexanoic acid N-hydroxysuccinimide which was first reacted either with 3-aminopropyltriethoxysilane or 3-aminopropyldimethylethoxysilane to form silane linkers 6-maleimide-N-(3-(triethoxysilyl)propyl)hexanamide or -(ethoxydimethylsilyl)propyl)-hexanamide. C-terminal cysteine genetically engineered to chimeric avidin was reacted with the maleimide group of silane linker in methanol/PBS solution to form a suspension, which was printed on sol-gel modified PMMA film. Different concentrations of chimeric avidin and ratios between silane linkers were tested to find the best properties for the bio-ink to enable gravure or inkjet printing. Bio-ink prepared from 3-aminopropyltriethoxysilane was found to provide the highest amount of active immobilized chimeric avidin. The developed bio-ink was shown to be valuable for automated fabrication of avidin-functionalized polymer films.

Disposable (bio)chemical integrated optical waveguide sensors implemented on roll-to-roll produced platforms

To enable wide spread dissemination of sensors in cost-critical applications and resource poor settings, methods to implement sensor chips using low-cost materials and mass-manufacturing methods are developed. In this paper we demonstrate that disposable polymeric integrated Young interferometer (YI) sensor chips, implemented on roll-to-roll mass-manufactured waveguides, are applicable for analyte specific sensing of small molecules and for multi-analyte detection of biomolecules. For the chemical sensing of small molecules, a sensor chip was functionalized with a molecularly imprinted polymer (MIP). We demonstrate that the MIP receptor layer is compatible with a polymer-based evanescent wave sensor for direct refractive index sensing. For multi-analyte detection of biomolecules, antibody-based receptor layers were patterned by inkjet printing onto the sensor surface demonstrating the applicability of the method with integrated YI chips. Demonstration of the analyte specific chemical- and biosensing with disposable polymeric YI sensor chips opens new possibilities to implement low-cost (bio)chemical sensors.

Printed hybrid systems

This paper presents research activities carried out at VTT Technical Research Centre of Finland in the field of hybrid integration of optics, electronics and mechanics. Main focus area in our research is the manufacturing of electronic modules and product structures with printed electronics, film-over-molding and polymer sheet lamination technologies and the goal is in the next generation of smart systems utilizing monolithic polymer packages. The combination of manufacturing technologies such as roll-to-roll -printing, injection molding and traditional component assembly is called Printed Hybrid Systems (PHS). Several demonstrator structures have been made, which show the potential of polymer packaging technology. One demonstrator example is a laminated structure with embedded LED chips. Element thickness is only 0.3mm and the flexible stack of foils can be bent in two directions after assembly process and was shaped curved using heat and pressure. The combination of printed flexible circuit boards and injection molding has also been demonstrated with several functional modules. The demonstrators illustrate the potential of origami electronics, which can be cut and folded to 3D shapes. It shows that several manufacturing process steps can be eliminated by Printed Hybrid Systems technology. The main benefits of this combination are small size, ruggedness and conformality. The devices are ideally suited for medical applications as the sensitive electronic components are well protected inside the plastic and the structures can be cleaned easily due to the fact that they have no joints or seams that can accumulate dirt or bacteria.

PDMS surface modification in the application of waveguide claddings for evanescent field sensing

We fabricated SU-8 based slab waveguides on surface-modified poly(dimethyl siloxane) PDMS lower claddings for application in evanescent field sensing. In this application, higher sensitivity is obtained by generating stronger penetrating power above the waveguide into the analyte. This can be achieved by reducing the refractive index of the substrate. Compared with glass substrates that have a refractive index of 1.5, PDMS has a refractive index of 1.42 at 633 nm, thus serving as a better lower cladding material for high-sensitivity sensing with an evanescent field or as claddings in multilayer waveguide applications. In order to increase the adhesion of PDMS surfaces for successful SU-8 application we treated PDMS thin films in low-frequency (40 kHz) oxygen plasma for varied length of exposure time. The treatment process made PDMS hydrophilic and created nano-structures on the surfaces. The resultant surface topography with different exposure time was studied by an interferometric profiler on PDMS lower claddings and the later spin-coated SU-8 waveguides. Measurement results showed that longer plasma treatment on PDMS claddings significantly improved the uniformity and waviness of the waveguides. Light propagation tests performed with a prism coupler and an end-butt coupling setup proved that PDMS can be used as a proper material for SU-8 waveguides.