This dissertation thesis describes a phenomenon called electroporation. It is about its theoretical aspects as well as about modeling of processes in the tissue during electroporation. Further, it describes the technical design of two developed unique experimental generators of DC and AC pulses for electroporation purposes. It also includes a description of experiments which were done using discussed generators.

The aim of this master thesis was to test and compare two available methods of genetic transformation (biolistics, electroporation) of the plastid genome of green algae Chlamydomonas reinhardtii. For biolistic transformation a wide range of experimental parameters which generally influence ptDNA transformation efficiency was optimized: physiological condition of acceptor cells, type and size of microparticles, pressure of propulsion gas (helium), length of projectile trajectory, transformation of cells directly on selective medium (containing spectinomycin 150 ?g/ml) or on nonselective medium (without spectinomycin) and form of transforming DNA. In contrast to nuclear transformation ptDNA transformation of the experimental object C. reinhardtii by means of electroporation hadn´t been described. Hence a wide range of values of different physical parameters which can significantly influence the transformation efficiency was tested.

The aim of this master thesis is to design a high voltage source for the generation of bipolar pulses, which are suitable for the process of reversible and irreversible electroporation. Further design is electronic control. The first part of the thesis analyzes the theoretical knowledge about the process of electroporation, the second part describes the principles of realization of such sources, the next part deals with the design of the source and its control. The penultimate part is a description of the user manual and the last part is dedicated to the measurement.

This masteral thesis describes phenomenon of electroporation and it’s use to deal with occluded self-expandable metalic stent. Thesis briefly summarizes theory of electroporation, currently used medical treatments of occluded stents. The next part of the thesis is dedicated to current state of mathematical simulations of electroporation. The core of the thesis are simulations of electroporation effects on tissue of occluded metal stents.

The aim of this master thesis is to design a power source for generating pulsed electric field for the needs of the technological process of electroporation of fruit musts and mashes. To design further the network of switched capacitors and inductors based on the required pulse and to design and implement the basic control unit together with the switching transistors. The thesis will include calculations and simulations used in the draft, also design solutions and measured values. This thesis is divided into several basic parts. In the first part there is discussed in detail the theoretical knowledge of electroporation and the desired characteristics of generated pulses are chosen according to this knowledge. Each part of electroporation workplace is described in the second part of this thesis, beginning from the source through the control system to the electrode chamber. For each of these parts are given different possible alternatives. In the next chapter is already proceeded to the design of the source. There are listed required parameters of the pulses and according to them calculations and the design are gradually carried out. Another chapter deals with the simulations, which are used to verify the calculated values and conditions in the electroporated sample. Last but one part discusses the mechanical design of the workplace. There are described all problems of the construction and commissioning of the product. The last section is dedicated to the workplace measurement and analysis of the measured results.

The aim of this work was to obtain better insight into the principles of cell structures and organs in the chicken embryo development. To reach this goal special methods of micromanipulations and visualization in vitro, ex vivo, ex ovo and in ovo were implemented and adjusted. These methods were used to study gene expression in neural crest development and eye development. In the course of long term research in our laboratory we observed that oncoprotein v-Myb influences the development of the neural crest and has the capacity to change natural cell fate. We performed a series of experiments to investigate v-Myb protein influence on neural crest cells differentiation, especially melanocyte lineage development, and its influence on gene expression in the neural crest. Therefore we focused on Gremlin 2 (PRDC), the gene upregulated by v-Myb in the neural crest. The established procedure of electroporation in ovo was adjusted to transfect cells of the developing eye and used to study gene expression during lens induction. The results obtained from chicken embryo experiments endorsed the study performed on mouse embryos. Futhermore, the electroporation technique was slightly modified for manipulations of the neural retina in the developing eye in ovo. Thereafter, the retinas were processed ex vivo and...

The main goal of this thesis is to design control board for the experimental electroporation device and to develop control firmware. The first chapter of this work focuses on the electroporation phenomenon itself. Behaviour of the cell model in external electrical field is described there as well as simulation and overview of how electroporation affects living tissue. It also explains the main requirements for parameters of the electroporation pulses as well as need for ECG synchronization. Furthermore, some remarks are given about novel high frequency electroporation method, which involves use of nanosecond bipolar high voltage pulse bursts. The second chapter briefly introduces commercial electroporation device called Nanoknife, including control part, power part, and it's limits. The third chapter consists of introduction of the novel experimental electroporation device developed at BUT. Power part of this device is discussed as well. Next chapter focuses on design of the control board for this device and also on description of the particular schematic parts. There is a control algorithm explanation in the fifth chapter of this thesis followed by the brief manual to machine operation.