Berlin 2015 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 1: Imaging

BP 1.1: Hauptvortrag

Montag, 16. März 2015, 09:30–10:00, H 1028

Light sheet-based fluorescence microscopy for quantitative biology — •Ernst H.K. Stelzer — Physical Biology, BMLS, Goethe Universität, D-60438 Frankfurt am Main

As long as we rely on epifluorescence microscopes, we are faced with serious challenges. Fluorophores and specimens are essentially wasted during the observation process, since all fluorophores and many endogenous organic compounds in the specimen are excited whenever we record a single plane. Obviously, the situation becomes even more challenging when we perform complex biological experiments and observe the behavior of multiple targets in three dimensions as a function of time. In light sheet-based fluorescence microscopy (LSFM), planar optical sectioning in the excitation process minimizes fluorophore bleaching and phototoxic effects. Since biological specimens survive long-term three-dimensional imaging at high spatio-temporal resolution, LSFM has become the tool of choice in developmental biology. LSFM makes a sincere and honest effort to reduce bleaching and phototoxicity. LSFM allows one to record millions of pixels in parallel. Laser light sheet-based devices, including a macroscope, had been built several times, but their capability to perform at a microscopic level was unknown until we described a diffraction-limited microscope in 2002, observed living biological samples and evaluated multiple-view imaging. This developed from theta microscopy (1993) and a systematic evaluation of diffraction-limited microscopes with two to four lenses, in theory and practice.