Zeitschriftenaufsätze

A major challenge in the use of HepG2 cell culture models for drug toxicity screening is their lack of maturity in 2D culture. 3D culture in Matrigel promotes the formation of spheroids that express liver-relevant markers, yet they still lack various primary hepatocyte functions. Therefore, alternative matrices where chemical composition and materials properties are controlled to steer maturation of HepG2 spheroids remain desired. Herein, a modular approach is taken based on a fully synthetic and minimalistic supramolecular matrix based on squaramide synthons outfitted with a cell-adhesive peptide, RGD for 3D HepG2 spheroid culture. Co-assemblies of RGD-functionalized squaramide-based and native monomers resulted in soft and self-recovering supramolecular hydrogels with a tunable RGD concentration. HepG2 spheroids are self-assembled and grown ( 150 m) within the supramolecular hydrogels with high cell viability and differentiation over 21 days of culture. Importantly, significantly higher mRNA and protein expression levels of phase I and II metabolic enzymes, drug transporters, and liver markers are found for the squaramide hydrogels in comparison to Matrigel. Overall, the fully synthetic squaramide hydrogels are proven to be synthetically accessible and effective for HepG2 differentiation showcasing the potential of this supramolecular matrix to rival and replace naturally-derived materials classically used in high-throughput toxicity screening.

The east and southeast rim of Harz mountains (Germany) are marked by a high density of former copper mining places dating back from the late 20th century to the middle age. A set of 18 soil samples from pre- and early industrial mining places and one sample from an industrial mine dump have been selected for investigation by 16S rRNA and compared with six samples from non-mining areas. Although most of the soil samples from the old mines show pH values around 7, RNA profiling reflects many operational taxonomical units (OTUs) belonging to acidophilic genera. For some of these OTUs, similarities were found with their abundances in the comparative samples, while others show significant differences. In addition to pH-dependent bacteria, thermophilic, psychrophilic, and halophilic types were observed. Among these OTUs, several DNA sequences are related to bacteria which are reported to show the ability to metabolize special substrates. Some OTUs absent in comparative samples from limestone substrates, among them Thaumarchaeota were present in the soil group from ancient mines with pH > 7. In contrast, acidophilic types have been found in a sample from a copper slag deposit, e.g., the polymer degrading bacterium Granulicella and Acidicaldus, which is thermophilic, too. Soil samples of the group of pre-industrial mines supplied some less abundant, interesting OTUs as the polymer-degrading Povalibacter and the halophilic Lewinella and Halobacteriovorax. A particularly high number of bacteria (OTUs) which had not been detected in other samples were found at an industrial copper mine dump, among them many halophilic and psychrophilic types. In summary, the results show that soil samples from the ancient copper mining places contain soil bacterial communities that could be a promising source in the search for microorganisms with valuable metabolic capabilities.

Defocus particle tracking (DPT) has gained increasing importance for its use to determine particle trajectories in all three dimensions with a single-camera system, as typical for a standard microscope, the workhorse of todays ongoing biomedical revolution. DPT methods derive the depth coordinates of particle images from the different defocusing patterns that they show when observed in a volume much larger than the respective depth of field. Therefore it has become common for state-of-the-art methods to apply image recognition techniques. Two of the most commonly and widely used DPT approaches are the application of (astigmatism) particle image model functions (MF methods) and the normalized cross-correlations between measured particle images and reference templates (CC methods). Though still young in the field, the use of neural networks (NN methods) is expected to play a significant role in future and more complex defocus tracking applications. To assess the different strengths of such defocus tracking approaches, we present in this work a general and objective assessment of their performances when applied to synthetic and experimental images of different degrees of astigmatism, noise levels, and particle image overlapping. We show that MF methods work very well in low-concentration cases, while CC methods are more robust and provide better performance in cases of larger particle concentration and thus stronger particle image overlap. The tested NN methods generally showed the lowest performance, however, in comparison to the MF and CC methods, they are yet in an early stage and have still great potential to develop within the field of DPT.

Nanomeasuring machines developed at the Technische Universität Ilmenau enable three-dimensional measurements and manufacturing processes with the lowest uncertainties. Due to the requirements for these processes, a highly reproducible and long-term stable tool changing system is needed. For this purpose, kinematically determined couplings are widely used. The state-of-the-art investigations on those are not sufficient for the highest demands on the reproducibility required for this application. A theoretical determination of the reproducibility based on analytical or numerical methods is possible, however not in the desired nanometer range. Due to this, a measurement setup for the determination of the reproducibility in five degrees of freedom with nanometer uncertainty was developed. First, potential measuring devices are systematically examined and measurement principles were developed out of this. A three-dimensional vector-based uncertainty analysis is performed to prove the feasibility of the measurement principle and provides a basis for further design. As a result, a translatory measurement uncertainty of 10 nm and a rotatory uncertainty of 11 nrad can be reached. Afterwards, the measurement setup is designed, focusing on the metrological frame and the lift-off device. The developed setup exceeds the uncertainties of the measurement setups presented in the state-of-the-art by an order of magnitude, allowing new in-depth investigations of the reproducibility of kinematic couplings.

Illumination with LEDs is of increasing interest in imaging and lithography. In particular, compared to lasers, LEDs are temporally and spatially incoherent, so that speckle effects can be avoided by the application of LEDs. Besides, LED arrays are qualified due to their high optical output power. However, LED arrays have not been widely used for investigating optical effects, e.g., the Lau effect. In this paper, we propose the application of an LED array for realizing the Lau effect by taking into account the influence of the coherence properties of illumination on the Lau effect. Using spatially incoherent illumination with the LED array or a single LED, triangular distributed Lau fringes can be obtained. We apply the obtained Lau fringes in the optical lithography to produce analog structures. Compared to a single LED, the Lau fringes using the LED array have significantly higher intensities. Hence, the exposure time in the lithography process is largely reduced.