Estimation of Injection Volume in Capillary Microinjection Using Electrical Impedance Measurement

Capillary pressure microinjection (CPM) is a tool for transporting small sample volumes into living cells utilizing a sharp glass pipette and pressure pulses. The automation level of the current state-of-the-art microinjection devices is low and this makes the technique slow, imprecise and inefficient. The objective of this thesis work is to develop a method to estimate the injection volume in the capillary pressure microinjection technique of living adherent cells. This method would improve the reliability and repeatability of CPM and facilitate automating the injection procedure. Due to the extremely small dimensions involved in the process, a straight measurement of the injection volume is not possible. The strategy used in this work is to generate a mathematical model for the injection volume as a function of the injection pressure and the pipette electrical resistance. A measurement setup is built around a microinjection system to gather data for constructing the model. The injection pressure is measured with a pressure sensor, the pipette electrical resistance is determined using a custom-made impedance measurement circuitry and the injection volume is estimated by using a fluorescent dye as the injection liquid and recording image data from the injections. Several injection pressures and micropipette sizes are used to achieve data extensively enough. A MATLAB based automated algorithm is generated to handle the measurement data and organize the results efficiently. The measurement results give a rough estimate of the relationship between the injection volume, the injection pressure and the pipette electrical resistance. However, a reliable model cannot be built based on the data. The reason is the rather limited amount of suitable measurement data for modelling it was possible to collect due to the numerous error situations. Nevertheless, new important information of the nature of the microinjection procedure is obtained and valuable observations on measurements connected to microinjection are made. Further studies must be done to solve the problems in the tests to be able to gather the data more efficiently and construct the actual model. /Kir1

Study of Speed and Force in Biomanipulation

Ph.DDOCTOR OF PHILOSOPH

Mechanical Manipulation and Characterization of Biological Cells

Mechanical manipulation and characterization of an individual biological cell is currently one of the most exciting research areas in the field of medical robotics. Single cell manipulation is an important process in intracytoplasmic sperm injection (ICSI), pro-nuclei DNA injection, gene therapy, and other biomedical areas. However, conventional cell manipulation requires long training and the success rate depends on the experience of the operator. The goal of this research is to address the drawbacks of conventional cell manipulation by using force and vision feedback for cell manipulation tasks. We hypothesize that force feedback plays an important role in cell manipulation and possibly helps in cell characterization. This dissertation will summarize our research on: 1) the development of force and vision feedback interface for cell manipulation, 2) human subject studies to evaluate the addition of force feedback for cell injection tasks, 3) the development of haptics-enabled atomic force microscope system for cell indentation tasks, 4) appropriate analytical model for characterizing the mechanical property of mouse embryonic stem cells (mESC) and 5) several indentation studies on mESC to determine the mechanical property of undifferentiated and early differentiating (6 days under differentiation conditions) mESC. Our experimental results on zebrafish egg cells show that a system with force feedback capability when combined with vision feedback can lead to potentially higher success rates in cell injection tasks. Using this information, we performed experiments on mESC using the AFM to understand their characteristics in the undifferentiated pluripotent state as well as early differentiating state. These experiments were done on both live as well as fixed cells to understand the correlation between the two during cell indentation studies. Our results show that the mechanical property of undifferentiated mESC differs from early differentiating (6th day) mESC in both live and fixed cells. Thus, we hypothesize that mechanical characterization studies will potentially pave the way for developing a high throughput system with force feedback capability, to understand and predict the differentiation path a particular pluripotent cell will follow. This finding could also be used to develop improved methods of targeted cellular differentiation of stem cells for therapeutic and regenerative medicine

Biologization, Nanotechnology, Simulation: Proceedings of the 1st Joint PhD Conference on Material Science:: from 27.6.-1.7.2022 in Dresden/ Germany and Usti/Česká republika

Materials scientists from Ústí nad Labem and Dresden met in June of 2022 for the first joint PhD Conference on Material Science, with the special focus on biologization, nanotechnology and simulation. The conference aimed to encourage interdisciplinary exchange between Čzech and German research institutes and promote transnational cooperation on an international level along the Saxon- Čzech border. Due to the restrictions caused by the corona pandemic, several attempts were necessary before the conference, which was first planned in 2020, could finally take place for the first time in 2022. The conference could take place in presence, which was seen as a big plus by all participants, especially as it was the first meeting in this German - Čzech context for most of the participants. The attending scientists (about 60) met at the Institute of Material Science of TU Dresden in Germany for the first half of the week before the conference moved to the faculties of Science and Environment of the Jan Evangelista Purkyně University UJEP in Ústí nad Labem in Čzechia. The organized activities ranged from scientific presentations of current PhD projects and research topics, lab tours in the participating institutions, come-together events such as a guided tour at the dye collection of the TU Dresden and a hiking trip to Bohemian Switzerland. The conference was funded by INTERREG VA Saxony - Čzech Republic - a cooperation programme of the Elbe/Labe region. All participants - PhD students, scientists and staff members of the participating institutions - enjoyed this opportunity to build individual and new contacts, exchange information on current research topics and methods, find starting points for future collaborations between the different research areas and institutions and also discuss the similarities and differences between the German and Čzech research landscape. The purpose of this brochure is to present the institutions with their special topics and laboratories and to present current research topics - on the base of the presented PhD projects.:1 Introduction 2 1.1 Committees 5 2 Presentation of the participating institutes and chairs 5 2.1 Jan Evangelista Purkyně University in Ústí nad Labem 6 2.1.1 Faculty of Science 6 2.1.2 Faculty of Environment 12 2.2 Technische Universität Dresden 17 2.2.1 Institute of Material Science 17 2.3 Fraunhofer Institute for Ceramic Technologies and Systems IKTS 19 2.3.1 Department Bio- and Nanotechnology at IKTS 19 2.4 Institute for Complex Materials, Leibniz-IFW Dresden 21 2.5 TRANS³Net 22 3 Presentation of the PhD topics 23 3.1 Topic: BIOLOGIZATION 23 3.1.1 Ludovico Andrea Alberta: Exploring the effect of Cu additions on the mechanical behaviour of β-TiNb biomaterials 23 3.1.2 Franziska Alt: Formation of a microenvironment for directed differentiation of stem cells in a perfusion bioreactor 25 3.1.3 Dmitry Belyaev: Circular microfluidic systems for electro-chemical continuous monitoring of bio-chemicals in emulsion droplets 27 3.1.4 Constantin Ißleib: Dynamic osteoimmunological crosstalk in a bone replacement context 28 3.1.5 Adela Jagerová: Surface Modification by High-Energy Heavy-Ion Irradiation in Various Crystalline ZnO Facets 29 3.1.6 Nils Kaube: Bioinspired development of artificial enamel via in-situ nano-mineralization 30 3.1.7 Michaela Kocholata: Isolation and characterization of plant derived nanovesicles 30 3.1.8 Zuzana Nejedlá: Dendrimers as Drug Delivery System 31 3.1.9 Jacub Perner: Effect of cold plasma treatment of Poppy and Proso Millet seeds in plasma downer 32 3.1.10 Marina Roshchina: Development of new bacteria-killing coatings on beta-Ti-Nb alloy based on functional oxide nanotubular (ONT) layers 33 3.1.11 Muhammad Saqib: Algorithms and fluid-dynamic experimental platform for in vitro degradation studies of implant materials 34 3.1.12 Jacub Tolasz: Interaction of pollutants on nanoceria 35 3.1.13 Zuzana Žmudová: 3D spheroid culture for in vitro testing of nanoparticles 35 3.2 Topic: METROLOGY 37 3.2.1 Katrien Boonen: The potential of dendrochemistry and dendroecology in pollution research 37 3.2.2 Ivan Lopez Carasco: Development of immobilization protocols for Tro6 and Tro4 aptamers to be used in electrochemical biosensor 38 3.2.3 Jacub Hoskovec: Functionalized electrospun materials for selectvie capture of selected gases 39 3.2.4 Dominic Pilnaj: Applications of gas sensors for air-quality monitoring and identification of volatile organic compounds by GC-HRMS 39 3.2.5 Michaela Průšová: Prostat, Glioblastoma and Mammary carcinoma cells derived exosomes: Their isolation, characterization and loading with doxorubicin 40 3.2.6 Kateřina Přibylová: Preparation of nanostructured surfaces for CO2 Detection, Capture and Utilization 41 3.2.7 Michal Syrový: Chemical modification of PAN – based nanofibrous membranes prepared by electrospinning and their properties for CO2 capture potential 42 3.3 Topic: GEOLOGICAL/MATERIALS 43 3.3.1 Sabine Apelt: Using biomimicry to design anti-ice surfaces for air-water heat pumps 43 3.3.2 Jan Dočkal: Molecular dynamics of interfacial solution structure of alkali-halide electrolytes at graphens electrodes 47 3.3.3 Tereza Dušková: Metal complexes with polyfluorinated NHCs 48 3.3.4 Kristína Fiantoková: Obtaining of the active mass from the spent Li-Ion batteries 48 3.3.5 Stephanie Ihmann: Engineering of bio-based Building and Construction Materials 49 3.3.6 Sara Jalali: Degradable bone substitute materials with load-bearing properties - Fiber-strengthened silica 50 3.3.7 Pavel Kaule: Preparation of heteroborane derivatives for thin film deposition by the covalent bond formation 53 3.3.8 M. Kozakovic: The effect of primary and secondary flows on the homogenization process in a vertical bladed mixer 53 3.3.9 Pavlína Matysová: Molecular Simulation of Salt Hydrates 54 3.3.10 Viktorie Neubertová: Surface functionalization of Ti3C2T MXene for MRI contrast agent 55 3.3.11 Robert Ato Newton: Fuel characteristics of Miscanthus x giganteus biomass produced at the marginal and slightly contaminated by trace elements soils 55 3.3.12 Martin Otto: Bioresorbable Fe-based alloys processed via laser powder bed fusion 56 3.3.13 Petr Panuška: A millifluidic chip for cultivation of fish embryos and toxicity testing fabricated by 3D printing technology 59 3.3.14 David Poustka: Unlocking mass production of photocrosslinked chitosan nanofibers 60 3.3.15 Eliška Rezlerová: Adsorption and Diffusion of Short Hydrocarbons and Carbon Dioxide in Shale Organic Matter: Insights from Molecular Simulations 60 3.3.16 Stefan Weitz: Investigating the material hardness of mollusks shells in dry and wet states by microindentation 6

The Impact of Food Bioactives on Health: In Vitro and Ex Vivo Models

Food Microbiology; Food Science; Human Physiolog

Multi-purpose impedance-based measurement system to automate microinjection of adherent cells

This paper presents a multi-purpose measurement system developed to automate cellular microinjection. The system is based on measuring impedance between an injection solution in an injection capillary and cell culture medium where cells grow. The system can be used for detecting a contact between a cell and the injection capillary, a broken capillary, a clogged capillary, an aged measurement electrode and faulty injection solution. The measurement system facilitates the development of an expert system for guiding cellular injections, and later a fully automatic microinjection system.Peer reviewe