Forschungszentrum Mikrotechnik
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Recently the use of microRNAs (miRNAs) as biomarkers for a multitude of diseases has gained substantial significance for clinical as well as point-of-care diagnostics. Amongst other challenges, however, it holds the central requirement that the concentration of a given miRNA must be evaluated within the context of other factors in order to unambiguously diagnose one specific disease. In terms of the development of diagnostic methods and devices, this implies an inevitable demand for multiplexing in order to be able to gauge the abundance of several components of interest in a patient’s sample in parallel. In this study, we design and implement different multiplexed versions of our electrochemical microfluidic biosensor by dividing its channel into subsections, creating four novel chip designs for the amplification-free and simultaneous quantification of up to eight miRNAs on the CRISPR-Biosensor X (‘X’ highlighting the multiplexing aspect of the device). We then use a one-step model assay followed by amperometric readout in combination with a 2-minute-stop-flow-protocol to explore the fluidic and mechanical characteristics and limitations of the different versions of the device. The sensor showing the best performance, is subsequently used for the Cas13a-powered proof-of-concept measurement of two miRNAs (miRNA-19b and miRNA-20a) from the miRNA-17∼92 cluster, which is dysregulated in the blood of pediatric medulloblastoma patients. Quantification of the latter, alongside simultaneous negative control measurements are accomplished on the same device. We thereby confirm the applicability of our platform to the challenge of amplification-free, parallel detection of multiple nucleic acids.
In this work, we present a significant step toward in vivo ophthalmic optical coherence tomography and angiography on a photonic integrated chip. The diffraction gratings used in spectral-domain optical coherence tomography can be replaced by photonic integrated circuits comprising an arrayed waveguide grating. Two arrayed waveguide grating designs with 256 channels were tested, which enabled the first chip-based optical coherence tomography and angiography in vivo three-dimensional human retinal measurements. Design 1 supports a bandwidth of 22 nm, with which a sensitivity of up to 91 dB (830 µW) and an axial resolution of 10.7 µm was measured. Design 2 supports a bandwidth of 48 nm, with which a sensitivity of 90 dB (480 µW) and an axial resolution of 6.5 µm was measured. The silicon nitride-based integrated optical waveguides were fabricated with a fully CMOS-compatible process, which allows their monolithic co-integration on top of an optoelectronic silicon chip. As a benchmark for chip-based optical coherence tomography, tomograms generated by a commercially available clinical spectral-domain optical coherence tomography system were compared to those acquired with on-chip gratings. The similarities in the tomograms demonstrate the significant clinical potential for further integration of optical coherence tomography on a chip system.
Electric cell-substrate impedance spectroscopy (ECIS) enables non-invasive and continuous read-out of electrical parameters of living tissue. The aim of the current study was to investigate the performance of interdigitated sensors with 50 μm electrode width and 50 μm inter-electrode distance made of gold, aluminium, and titanium for monitoring the barrier properties of epithelial cells in tissue culture. At first, the measurement performance of the photolithographic fabricated sensors was characterized by defined reference electrolytes. The sensors were used to monitor the electrical properties of two adherent epithelial barrier tissue models: renal proximal tubular LLC-PK1 cells, representing a normal functional transporting epithelium, and human cervical cancer-derived HeLa cells, forming non-transporting cancerous epithelial tissue. Then, the impedance spectra obtained were analysed by numerically fitting the parameters of the two different models to the measured impedance spectrum. Aluminium sensors proved to be as sensitive and consistent in repeated online-recordings for continuous cell growth and differentiation monitoring assensors made of gold, the standard electrode material. Titanium electrodes exhibited an elevated intrinsic ohmic resistance incomparison to gold reflecting its lower electric conductivity. Analysis of impedance spectra through applying models and numerical data fitting enabled the detailed investigation of the development and properties of a functional transporting epithelial tissue using either gold or aluminium sensors. The result of the data obtained, supports the consideration of aluminium and titanium sensor materials as potential alternatives to gold sensors for advanced application of ECIS spectroscopy.
An electrochemical study with three redox substances on a carbon based nanogap electrode array
(2020)
Over the last years, polymers have gained great attention as substrate material, because of the possibility to produce low-cost sensors in a high-throughput manner or for rapid prototyping and the wide variety of polymeric materials available with different features (like transparency, flexibility, stretchability, etc.). For almost all biosensing applications, the interaction between biomolecules (for example, antibodies, proteins or enzymes) and the employed substrate surface is highly important. In order to realize an effective biomolecule immobilization on polymers, different surface activation techniques, including chemical and physical methods, exist. Among them, plasma treatment offers an easy, fast and effective activation of the surfaces by micro/nanotexturing and generating functional groups (including carboxylic acids, amines, esters, aldehydes or hydroxyl groups). Hence, here we present a systematic and comprehensive plasma activation study of various polymeric surfaces by optimizing different parameters, including power, time, substrate temperature and gas composition. Thereby, the highest immobilization efficiency along with a homogenous biomolecule distribution is achieved with a 5-min plasma treatment under a gas composition of 50% oxygen and nitrogen, at a power of 1000 W and a substrate temperature of 80 C. These results are also confirmed by different surface characterization methods, including SEM, XPS and contact angle measurements.
We present a new concept of 3D polymer-based 1 × 4 beam splitter for wavelength splitting around 1550 nm. The beam splitter consists of IP-Dip polymer as a core and polydimethylsiloxane (PDMS) Sylgard 184 as a cladding. The splitter was designed and simulated with two different photonics tools and the results show high splitting ratio for single-mode and multi-mode operation with low losses. Based on the simulations, a 3D beam splitter was designed and realized using direct laser writing (DLW) process with adaptation to coupling to standard single-mode fiber. With respect to the technological limits, the multi-mode splitter having core of (4 × 4) μm 2 was designed and fabricated together with supporting stable mechanical construction. Splitting properties were investigated by intensity monitoring of splitter outputs using optical microscopy and near-field scanning optical microscopy. In the development phase, the optical performance of fabricated beam splitter was examined by splitting of short visible wavelengths using red light emitting diode. Finally, the splitting of 1550 nm laser light was studied in detail by near-field measurements and compared with the simulated results. The nearly single-mode operation was observed and the shape of propagating mode and mode field diameter was well recognized.
Design and optimization of 1x2N Y-branch optical splitters for telecommunication applications
(2020)
This paper presents the design and optimization of 1x2N Y-branch optical splitters for telecom applications. A waveguide channel profile, used in the splitter design, is based on a standard silica-on-silicon material platform. Except for the lengths of the used Y-branches, design parameters such as port pitch between the waveguides and simulation parameters for all splitters were considered fixed. For every Y-branch splitter, insertion loss, non-uniformity, and background crosstalk are calculated. According to the minimum insertion loss and minimum non-uniformity, the optimum length for each Y-branch is determined. Finally, the individual Y-branches are cascade joined to design various Y-branch optical splitters, from 1x2 to 1x64.
