Fachbereich Ingenieur- und Naturwissenschaften
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The novel approach of an individualized medicine affects nowadays various areas of therapeutic treatment. Primarily, these include the application of pharmaceuticals, private point-of-care solutions, surgical procedures as well as steps of rehabilitation. However, beside these existing strategies the development of patient specific models for the training of clinical personal is currently insufficient. Such models are essential to prepare a personalized approach of medical care. One possible solution to address this problem could be the adaption of the established 3d printing technology for the processing of suitable biopolymers. The presented work is focused on the development of a printing system utilizing collagen. Therefore, it is intended to design multiple extrusion heads in accordance with the material parameters. In order to achieve the aspired physiological properties of the final model, a first set of experiments will be performed with several compositions of collagen to validate the fundamental mechanical characteristics. Hereby, the elasticity, thermal stability, force resistance as well as the haptic behaviour are of most interest. Afterwards, these obtained experimental results should be used to simulate the extrusion process and to validate the extruder concepts. In case of a positive evaluation these concepts are realized by using rapid prototyping technologies. Finally, this novel 3d printer will be used to print first organic test structures with collagen.
The sample preparation for biological and chemical probes involves following a strict workflow to eliminate any contamination to the sample beforehand. Furthermore, it is time consuming and must be carried out by trained personnel such as a nurse or other supervisors, making it therefore expensive. The development of novel sample preparation techniques paired with modern sample analysis systems is focused on improving the operability while keeping a constant quality of results. This is important to analyse samples, which cannot be determined with current screening conditions. The analysis of analytes is required to receive a more detailed picture of the patient and to fully understand its complexity. Possible samples for in-depth analysis of chemical origin can be cholesterol or glucose. More complex samples, such as blood or saliva, require a sophisticated system, which analyses the samples for their individual compounds.
In-vitro cell culture offers the ability to grow individual cells and monitor their behaviour in a controlled environment over a certain time. During culture, different parameters have to be controlled to ensure a vital growth of cells. The most important parameters are temperature, pH-value, oxygen and carbon dioxide levels as well as the glucose concentration. All of these parameters influence the growth ability of the cell culture and should be monitored online. The online monitoring of glucose was achieved with a novel GOD based sensor which enabled a real-time measurement of glucose during cell culture. A first-time characterisation of the sensor was carried out in the culture medium DMEM. In addition, material properties of the culture system were investigated. At first to establish a baseline measurement, the sensor was placed in different glucose concentrations dissolved in Water-KCl mixture at 25 °C inside a batch chamber under constant stirring. Afterwards, the temperature was increased to 37 °C to simulate a cell culture environment. In addition, the system was transferred into a flow-through reaction chamber. The highest response signal and the most stable signal was achieved at 37 °C with DMEM during flow-through measurement. Based on these measurements online glucose monitoring in cell culture was possible to determine the glucose consumption for each cell which is important for future human-on-a-chip devices to prevent diabetic metabolisms.
Modern medical science delivers through innovative chemical or mechanical/physical means new strategies to treat patients mildly and fight diseases accurately. In line with this development a screening procedure for tissue samples under usage of the electronic speckle pattern interferometry is developed at the University of Applied Science Wildau. The paper at hand provides the corner stone for such a procedure in form of an incubation system that is adapted to the properties of an electronic speckle pattern interferometer and allows the incubation as well as study of samples over time. As a result the developed system can regulate its own temperature and is constructed for use in an electronic speckle pattern interferometry (ESPI) setup. Its design allows a simple modular approach for further development.
3 dimensional (3D) printing evolved during the last decade to a consumer friendly and affordable craft. Furthermore, implementations of this techniques in the field of biotechnological research and development within laboratories is a very expansive process. Bio-printers’ prices cover a wide spectrum and most basic models are available for around 5000€. On the other end, high-end printer machines with a vast variety of features are available for several hundreds of thousands of euros. Thus, due to the immense potential in the field of Biotechnology the availability of this technology for research purpose should be enhanced. A developed ecological syringe extruder prototype for processing of biological based gels has been further improved. The original prototype was capable to processing multiple layers of agar with concentrations of 1% and 2.5%. Based on these results the prototype was revised regarding printing process parameter, which include among others applied forces to the substrate, air-ventilation, and heating of the substrate. The process behavior will be simulated with computational fluid dynamics for the processing of biological based substrate. After a concluding validation these results are intended to be implemented into a new design for improved processing of a variety of bioinks.
Bioreactor systems for cultivating cells in Life Sciences have been widely used for decades. Recently, there is a trend towards miniaturization, disposables and even micro platforms that fulfill increasing demands strongly aiming for production and testing of novel pharmaceutical products. Miniaturized bioreactors allow low power consumption, portability and reduced space requirements and utilize smaller volumes of reagents and samples [1,2]. A recursive strategy is necessary for optimizing the design and the manufacture of such miniaturized bioreactors. For the fabrication of these prototypes utilized micro-milling. Micro milling is a mechanical process which is commonly applied to create micro-structures in metals, e.g. aluminum and steel, or polymers, e.g. poly carbonate substrates. The structures and geometries are generated by utilizing computer aided design. By means of computer-aided manufacturing, the machining operations are implemented and then transferred to the machine tool. The machine tool moves the cutting tools with certain speeds, feeds and traverse ranges to the substrate. Micro milling has the advantage that the materials are generally not degraded by chemical substances, heating procedures or electromagnetic radiation.
Development of a Smartphone Based Reader for the Quantitative Analysis of Lateral Flow Assays
(2018)
Smartphones are developing into all-purposes devices. In the present work, the employment/application of smartphones as medical devices in home care and point-of-care (POC) diagnostics are investigated in the analysis of Lateral Flow Assays (LFA). A smartphone-based LFA reader was developed for the quantitative analysis of D-Dimer – a biomarker indicating e.g. thrombotic event or danger of embolism.The proof-of-concept has been shown with multiple smartphones in establishing: (I) Optimal dimensions of the LFA cell of 72.11mm distance of smartphone to D-Dimer test leading to a coefficients of variances (CV) between 0.8% and 4.2%. (II) Inter-device investigations: CVs around 13.5%; a limit of detection (LOD) of 100ng/ml (DDU) D-Dimer. (III) Inter-smartphone investigations: CV about 16%, a limit of detection (LOD) at 66.4ng/ml (DDU). (IV) Calibrations: CV and LOD of three smartphones are comparable to the commercial available LFA reader. Further development to put the multiple smartphone-based LFA reader on the market.
Modern cell culture as well as sophisticated bio-applications involve complex biochemical processes, which are required to induce growth, product development or material degradation. Tracking the reaction processes inside the application presents a major challenge due to its complexity. The development of new analysis and tracking mechanisms for such application presents a solution to fully understand the process. In addition, the applied sensors are required to monitor the reactions enable a live tracking of the process. Furthermore, this gives the opportunity to influence and manipulate reactions to further enhance the application of the process. Possible analytes for tracking during processes can be chemical origin such as glucose, cytokines, antibiotics and growth factors, which are included in the culture medium. Based on the complexity of the culture or bio-application the sensor tracking mechanism has to be adapted to ensure full process control. A variety of different approaches can be used for the tracking mechanism.
BACKGROUND:
After excitation with light photoacids can change the pH in a solution by release of a proton. They have been used mostly for excited state proton transfer studies. In this review the general functionality and mechanisms and the subdivision of photoacids is explained.
STATE OF THE ART:
Different uses of photoacids are described, covering a wide range of various biochemical topics, focusing on biochemical applications. Examples for the introduced subdivisions are covered.
CONCLUSIONS AND OUTLOOK:
The areas in which photoacids can be employed are diverse. Photoacids have a promising future in biotechnology and biochemistry and should be considered for upcoming applications, especially in non-invasive control of biochemical reactions.
The binding of photosystem I (PS I) from Thermosynechococcus elongatus to the native cytochrome (cyt) c6 and cyt c from horse heart (cyt cHH) was analyzed by oxygen consumption measurements, isothermal titration calorimetry (ITC), and rigid body docking combined with electrostatic computations of binding energies. Although PS I has a higher affinity for cyt cHH than for cyt c6, the influence of ionic strength and pH on binding is different in the two cases. ITC and theoretical computations revealed the existence of unspecific binding sites for cyt cHH besides one specific binding site close to P700. Binding to PS I was found to be the same for reduced and oxidized cyt cHH. Based on this information, suitable conditions for cocrystallization of cyt cHH with PS I were found, resulting in crystals with a PS I:cyt cHH ratio of 1:1. A crystal structure at 3.4-Å resolution was obtained, but cyt cHH cannot be identified in the electron density map because of unspecific binding sites and/or high flexibility at the specific binding site. Modeling the binding of cyt c6 to PS I revealed a specific binding site where the distance and orientation of cyt c6 relative to P700 are comparable with cyt c2 from purple bacteria relative to P870. This work provides new insights into the binding modes of different cytochromes to PS I, thus facilitating steps toward solving the PS I–cyt c costructure and a more detailed understanding of natural electron transport processes.