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DDC: 500 × Subject: imaging ×

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    Nonlinear Structured Illumination Using a Fluorescent Protein Activating at the Readout Wavelength
    (San Francisco, California, US : PLOS, 2016) Lu-Walther, Hui-Wen; Hou, Wenya; Kielhorn, Martin; Arai, Yoshiyuki; Nagai, Takeharu; Kessels, Michael M.; Qualmann, Britta; Heintzmann, Rainer
    Structured illumination microscopy (SIM) is a wide-field technique in fluorescence microscopy that provides fast data acquisition and two-fold resolution improvement beyond the Abbe limit. We observed a further resolution improvement using the nonlinear emission response of a fluorescent protein. We demonstrated a two-beam nonlinear structured illumination microscope by introducing only a minor change into the system used for linear SIM (LSIM). To achieve the required nonlinear dependence in nonlinear SIM (NL-SIM) we exploited the photoswitching of the recently introduced fluorophore Kohinoor. It is particularly suitable due to its positive contrast photoswitching characteristics. Contrary to other reversibly photoswitchable fluorescent proteins which only have high photostability in living cells, Kohinoor additionally showed little degradation in fixed cells over many switching cycles.
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    Two-dimensional imaging in hyperbolic media-the role of field components and ordinary waves
    ([London] : Macmillan Publishers Limited, part of Springer Nature, 2015) Tuniz, Alessandro; Kuhlmey, Boris T.
    We study full vector imaging of two dimensional source fields through finite slabs of media with extreme anisotropy, such as hyperbolic media. For this, we adapt the exact transfer matrix method for uniaxial media to calculate the two dimensional transfer functions and point spread functions for arbitrary vector fields described in Cartesian coordinates. This is more convenient for imaging simulations than the use of the natural, propagation direction-dependent TE/TM basis and clarifies which field components contribute to sub-diffraction imaging. We study the effect of ordinary waves on image quality, which previous one-dimensional approaches could not consider. Perfect sub-diffraction imaging can be achieved if longitudinal fields are measured, but in the more common case where field intensities or transverse fields are measured, ordinary waves cause artefacts. These become more prevalent when attempting to image large objects with high resolution. We discuss implications for curved hyperbolic imaging geometries such as hyperlenses.