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Dye Distance Mapping Using Waveguide Evanescent Field Fluorescence Microscopy And Its Application To Cell Biology, Frederik Fleissner, Michael Morawitz, S. Jeffrey Dixon, Uwe Langbein, Silvia Mittler

Physics and Astronomy Publications

Previous studies have measured the distance between cells and the substratum at sites of adhesion via the emission of a fluorescent dye and waveguide methods. Here, we demonstrate a novel approach to measure the position of fluorescent dyes above a waveguide surface in the 10-200 nm distance range throughout an entire area, yielding a 2D dye distance map or a 3D contour plot. The dye is located in a multilayered Langmuir Blodgett (LB) film or in the plasma membrane of a cell. Waveguide evanescent field fluorescence (WEFF) images obtained using two different waveguide modes are employed allowing, with a simple …