title: Nonlinear Structured Illumination Using a Fluorescent Protein Activating at the Readout Wavelength.
abstract: 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.
authors: Leibniz Institute of Photonic Technology, Jena, Germany.
authors: Hui-Wen
authors: http://orcid.org/0000-0002-8710-2100
authors: HW
authors: Lu-Walther
authors: Institute of Biochemistry I, Jena University Hospital/Friedrich Schiller University Jena, Jena, Germany.
authors: Wenya
authors: W
authors: Hou
authors: Leibniz Institute of Photonic Technology, Jena, Germany.
authors: Martin
authors: M
authors: Kielhorn
authors: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors: Yoshiyuki
authors: Y
authors: Arai
authors: The Institute of Scientific and Industrial Research, Osaka University, Osaka, Japan.
authors: Takeharu
authors: T
authors: Nagai
authors: Institute of Biochemistry I, Jena University Hospital/Friedrich Schiller University Jena, Jena, Germany.
authors: Michael M
authors: MM
authors: Kessels
authors: Institute of Biochemistry I, Jena University Hospital/Friedrich Schiller University Jena, Jena, Germany.
authors: Britta
authors: B
authors: Qualmann
authors: Leibniz Institute of Photonic Technology, Jena, Germany.|Institute of Physical Chemistry, Abbe Center of Photonics, Friedrich-Schiller-University Jena, Jena, Germany.
authors: Rainer
authors: R
authors: Heintzmann
mesh_terms: D006367:HeLa Cells; D006801:Humans; D008027:Light; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D017711:Nonlinear Dynamics
chemical_list: D008164:Luminescent Proteins