Subdiffraction Imaging through the Selective Donut-Mode Depletion of Thermally Stable Photoswitchable Fluorophores: Numerical Analysis and Application to the Fluorescent Protein Dronpa.

The fast and reversible on/off switching of the fluorescence emission of the GFP-like fluorescent protein Dronpa has attracted considerable interest for applications in subdiffraction imaging. In this paper we study the use of a donut-mode beam in combination with two more overlapping laser beams to...

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Publicado en:Journal of the American Chemical Society Vol. 129; no. 51; pp. 16132 - 16142
Autores principales: Dedecker, Peter, Hotta, Jun-ichi, FIors, Cristina, Sliwa, Michel, Uji-i, Hiroshi, Roeffaers, Maarten B. J., Ando, Ryoko, Mizuno, Hideaki, Miyawaki, Atsushi, Hofkens, Johan
Formato: Artículo
Publicado: American Chemical Society 12/26/2007
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Acceso en línea:Ver este registro en EBSCOhost
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      dt: 12/26/2007
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        atl: Subdiffraction Imaging through the Selective Donut-Mode Depletion of Thermally Stable Photoswitchable Fluorophores: Numerical Analysis and Application to the Fluorescent Protein Dronpa.
      aug:
        au:
          Dedecker, Peter
          Hotta, Jun-ichi
          FIors, Cristina
          Sliwa, Michel
          Uji-i, Hiroshi
          Roeffaers, Maarten B. J.
          Ando, Ryoko
          Mizuno, Hideaki
          Miyawaki, Atsushi
          Hofkens, Johan
        affil:
          Department of Chemistry and INPAC, Katholieke Universiteit Leuven, Celestijnenlaan 200 F, 3001 Heverlee, Belgium
          Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan
      su:
        Numerical analysis
        Green fluorescent protein
        Fluorescence
        Laser beams
        Optical resolution
        Optical diffraction
      sug:
        subj:
          Numerical analysis
          Green fluorescent protein
          Fluorescence
          Laser beams
          Optical resolution
          Optical diffraction
      ab: The fast and reversible on/off switching of the fluorescence emission of the GFP-like fluorescent protein Dronpa has attracted considerable interest for applications in subdiffraction imaging. In this paper we study the use of a donut-mode beam in combination with two more overlapping laser beams to increase the imaging resolution through selective switching to the nonfluorescent photoswitched state. We devise and run a series of numerical simulations to determine suitable photophysical parameters of prospective, thermally stable photoswitchable molecules, in terms of photoswitching quantum yields, fatigue resistance, and possible presence of transient nonfluorescent states. Many of our findings are applicable to other measurements that make use of donut beams, and these guidelines can be used in the synthesis and screening of novel photoswitchable compounds. We experimentally demonstrate the possibility of obtaining increased resolution by making use of the efficient and thermally stable Dronpa photoswitching, using equipment that is commonly available.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: English
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