Fluorescence microscope (wide field system with inverse optics)

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Intracellular membranes subdivide eukaryotic cells into specific compartments which allows that different processes can occur simultaneously in a spatially separated and rather independent manner. For the biogenesis of cells, this compartmentalization, however, can be a big challenge for several reasons: (i) Eukaryotic cells contain two, in the case of algae and plants even three genomes from which gene expression needs to be coordinated. (2) Newly synthesized proteins have to be transported from the site of their synthesis to their final destination of function, for which, in most cases, they have to be translocated across membranes. (3) Comprehensive quality control mechanisms need to make sure that all these processes in the different compartments are coordinated for which it is required that local problems are immediately recognized and solved.In several research projects we analyze the biogenesis of eukaryotic cells, thereby particularly focusing on processes in genome-containing organelles such as mitochondrial and chloroplasts. We predominantly use the baker’s yeast Saccharomyces cerevisiae and the algae Chlamydomonas reinhardtii simple model organisms. For specific experiments, however, we also work with cultured mammalian cells.Here we apply for a fluorescence microscope with inverse optics for life cell imaging. This instrument will be used to localize simultaneously several individual fluorescence-labeled proteins in these very small cells and to study their intracellular distribution under different physiological and stress conditions. By use of microfluidics chambers the cells will be visualized for several generations, for example to elucidate the formation and inheritance of protein aggregates and to measure the dynamic form, volume and distribution organelles and their genomes within cells. In addition to an excellent optic system and fully automatization devices, software packages will be required for the analysis of the images, for example for deconvolution. From these projects, we expect detailed molecular insights into the function and relevance of specific chaperones, assembly factors, translocation complexes and proteases for the biogenesis of mitochondria and chloroplasts as well as for processes which maintain the functionality of eukaryotic cells under variable (patho)physiological conditions.