Berlin 2008 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 26: Posters II

BP 26.30: Poster

Donnerstag, 28. Februar 2008, 17:00–19:30, Poster A

Single molecule microscopy using total internal reflection — •Andreas Veenendaal, Jan Peter Siebrasse, Constanze Husche, and Ulrich Kubitscheck — Institute of Physical and Theoretical Chemistry, Bonn, Germany

In single molecule fluorescence microscopy one often wants to reduce the fluorescence background emitted by fluorophores not located in the plane of interest. With total internal reflection fluorescence (TIRF) microscopy this is achieved by reflecting the light at the coverslip/probe interface (coverslip (glass) n = 1.51; sample (e.g. cell) n = 1.33), and thus generating an evanescent wave illuminating the sample. The intensity of the evanescent wave decays exponentially with a penetration depth of roughly half the wavelength of the incident light. We realised objective type TIRF using an NA 1.45 objective from Zeiss. Using this technique we characterized several red fluorescent dyes attached to the surface by a PEG-biotin-streptavidin system at the single molecule level. As a first biological application we studied the transport dynamics of single nuclear pore complexes.