Universal Micromachining Platform and Basic Technologies for the Manufacture and Marking of Microphysiological Systems.

Micro Physiological Systems (MPS), also known as Multi-Organ-Chip, Organ-on-a-Chip, or Body-on-a-Chip, are advanced microfluidic systems that allow the cultivation of different types of cells and tissue in just one common circuit. Furthermore, they thus can also adjust the interaction of these different tissues. Perspectival MPS will replace animal testing. For fast and flexible manufacturing and marking of MPS, a concept for a universal micromachining platform has been developed which provides the following latest key technologies: laser micro cutting of polymer foils, laser micro- and sub-micro-structuring of polymer foils, 3D printing of polymer components as well as optical inspection and online process control. The combination of different laser sources, processing optics, inspection systems, and print heads on multiple axes allows the change and exactly positioning to the workpiece during the process. Therewith, the realization of MPS including 3D printed components as well as direct laser interference patterned surfaces for well-defined cell adhesion and product protection is possible. Additional basic technologies for the generation of periodical line-like structures at polycarbonate foils using special Direct Laser Interference Patterning (DLIP) optics as well as for the 3D printing of fluid-tight cell culture reservoirs made of Acrylonitrile Butadiene Styrene directly onto polycarbonate microfluidics were established. A first prototype of the universal micromachining platform combining different lasers with Direct Laser Writing and DLIP is shown. With this laser micro cutting as well as laser micro-structuring of polycarbonate (PC) foils and therewith functionalization for MPS application could be successfully demonstrated.

Ultrasensitive SPR detection of miRNA-93 using antibody-enhanced and enzymatic signal amplification.

MiRNAs are endogenous noncoding RNA molecules. They play important gene-regulatory roles by binding to the mRNA of target genes thereby leading to either transcript degradation or translational repression. In virtually all diseases, distinct alterations of miRNA expression profiles have been found thus suggesting miRNAs as interesting biomarkers. Here, we present an SPR biosensor that utilizes disposable, injection-molded sensor chip/microfluidic hybrids combined with a lateral imaging optical system for parallel analysis of three one-dimensional spot arrays to detect miRNA-93. To increase the sensitivity of the biosensor we used two different amplification strategies. By adding an RNA-DNA-hybrid antibody for primary signal amplification, a limit of detection of 10 pmol/L was achieved. Based on that method we demonstrate the detection of miRNA-93 in total RNA lysate from HEK-293 cells. Utilizing an enzymatic signal amplification with Poly(A) polymerase, the sensitivity could be increased even further leading to a limit of detection of 1 fmol/L.

Effective photodarkening suppression in Yb-doped fiber lasers by visible light injection.

Al-silicate fibers have excellent manufacturing quality. Unfortunately, high-Yb doping concentration may be limited by severe losses induced by photodarkening phenomenon. In this paper we demonstrate for the first time that Al-silicate Yb-doped fibers with high-inversion and doping concentration above 1 wt% can be successfully used by implementing a simple optical bleaching scheme. A co-injection into the active fiber of a few mW of light at around 550 nm wavelength successfully eliminates almost all photodarkening induced losses. We demonstrate operation at above 90% of the pristine output power level in several lasers with up to 30% Yb ions in the excited state. These results may allow using Yb-doped Al-silicate fibers with doping level increased by one order of magnitude. Finally, we provide a comprehensive picture of main parameters affecting photobleaching performance and, to the best of our knowledge, we report the first quantitative measurement of the Ytterbium excited state absorption cross-section in the visible range.

'Human-on-a-chip' developments: a translational cutting-edge alternative to systemic safety assessment and efficiency evaluation of substances in laboratory animals and man?

Various factors, including the phylogenetic distance between laboratory animals and humans, the discrepancy between current in vitro systems and the human body, and the restrictions of in silico modelling, have generated the need for new solutions to the ever-increasing worldwide dilemma of substance testing. This review provides a historical sketch on the accentuation of this dilemma, and highlights fundamental limitations to the countermeasures taken so far. It describes the potential of recently-introduced microsystems to emulate human organs in 'organ-on-a-chip' devices. Finally, it focuses on an in-depth analysis of the first devices that aimed to mimic human systemic organ interactions in 'human-on-a-chip' systems. Their potential to replace acute systemic toxicity testing in animals, and their inability to provide alternatives to repeated dose long-term testing, are discussed. Inspired by the latest discoveries in human biology, tissue engineering and micro-systems technology, this review proposes a paradigm shift to overcome the apparent challenges. A roadmap is outlined to create a new homeostatic level of biology in 'human-on-a-chip' systems in order to, in the long run, replace systemic repeated dose safety evaluation and disease modelling in animals.

DNA microarrays for hybridization detection by surface plasmon resonance spectroscopy.

We report on the development of a new platform technology for the detection of genetic variations by means of surface plasmon resonance (SPR) spectroscopy. TOPAS chips with integrated optics were exploited in combination with microfluidics. Within minutes, the detection of hybridization kinetics was achieved simultaneously at all spots of the DNA microarray. A nanoliter dispenser is used to deposit thiol-modified single-stranded probe DNA on the gold surface of the chips. We investigated the influence of different parameters on hybridization using model polymerase chain reaction (PCR) products. These PCR products comprised a single-stranded tag sequence being complementary to an anti-tag sequence of probes immobilized on the gold surface. The signals increased with increasing length of PCR products (60, 100 or 300 base pairs) as well as with their concentration. We investigated hybridizations on DNA microarrays comprising 90 spots of probe DNA with three different sequences. Furthermore, we demonstrate that sequences with possible hairpin structures significantly lower the binding rate, and thus, the SPR signals during hybridization.

Design and prototyping of a chip-based multi-micro-organoid culture system for substance testing, predictive to human (substance) exposure.

Dynamic miniaturized human multi-micro-organ bioreactor systems are envisaged as a possible solution for the embarrassing gap of predictive substance testing prior to human exposure. A rational approach was applied to simulate and design dynamic long-term cultures of the smallest possible functional human organ units, human "micro-organoids", on a chip the shape of a microscope slide. Each chip contains six identical dynamic micro-bioreactors with three different micro-organoid culture segments each, a feed supply and waste reservoirs. A liver, a brain cortex and a bone marrow micro-organoid segment were designed into each bioreactor. This design was translated into a multi-layer chip prototype and a routine manufacturing procedure was established. The first series of microscopable, chemically resistant and sterilizable chip prototypes was tested for matrix compatibility and primary cell culture suitability. Sterility and long-term human cell survival could be shown. Optimizing the applied design approach and prototyping tools resulted in a time period of only 3 months for a single design and prototyping cycle. This rapid prototyping scheme now allows for fast adjustment or redesign of inaccurate architectures. The designed chip platform is thus ready to be evaluated for the establishment and maintenance of the human liver, brain cortex and bone marrow micro-organoids in a systemic microenvironment.