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BP: Fachverband Biologische Physik

BP 2: Bioimaging and Spectroscopy I

BP 2.1: Hauptvortrag

Montag, 20. März 2017, 09:30–10:00, HÜL 386

Visualization and Manipulation of the Invisible — •Heinrich Leonhardt — Ludwig Maximilians University Munich, Biocenter, Martinsried, Germany

Fluorescence light microscopy allows multicolor visualization of cellular components with high specificity, but its utility has until recently been constrained by the intrinsic limit of spatial resolution and the lack of specific detection tools. To circumvent these limitations, we applied three-dimensional structured illumination microscopy (3D-SIM, Science, 320, 1332-6) and high-throughput STED microscopy in combination with automated image analysis. For detection of cellular structures, we have generated fluorescent, antigen-binding proteins, termed chromobodies, by combining epitope-recognizing fragments with fluorescent proteins (Nature Methods, 3, 887-9). These chromobodies can be expressed in living cells and used to target or trace epitopes in subcellular compartments providing an optical readout for novel high content analyses and functional studies (Nature Struct. Mol. Biol., 17, 133-139). These antigen-binding fragments can also be recombinantly produced, chemically functionalized and directly used for super-resolution microscopy (Science, 331, 1616-20). To study the dynamics of genome organization we have repurposed prokaryotic DNA binding proteins (TALEs and CRISPR/Cas) for the detection of specific DNA sequences in living cells (NAR 42, e38 and Nucleus, 5, 163-172). This combination of detection tools and microscopy techniques provides new insights into the structure and function of mammalian cells.