The topic of this thesis is the study of developmental biological processes using modern light microscopy techniques. The focus is on the embryogenesis of the nematode Caenorhabditis elegans. For this purpose, an already existing setup is refined, characterized and used for gentle three-dimensional long-term imaging of the early embryogenesis. The results of another thesis, which were made possible by these acquisitions, are briefly presented. An extension of the setup to perform diffusion measurements enables a comparison of three different measurement techniques (SPIM-FCS, SPT, DDM) using calibration samples. Based on these first results, a subsequent spatial and temporal quantification of protein diffusion in living embryos is performed. After first successful measurements on a peripheral membrane protein in the cytoplasm and on the membrane of the embryo, the development of a spatial diffusion gradient of a protein, which is critical for the embryonic development, is examined. In a subsequent part of this thesis the development, construction and characterization of a second setup for super-resolution microscopy as well as bleaching and ablation measurements is presented. In addition to the gain of resolution these techniques facilitate both the quantification of diffusion and binding processes and a minimally invasive method for specific sample manipulations. As a result of this, new issues and relevant key processes in the embryonic development can be explored. The methods are tested on culture cells and worm embryos and first results are presented.
|School:||Universitaet Bayreuth (Germany)|
|Source:||DAI-C 81/1(E), Dissertation Abstracts International|
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