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      Fast, background-free, 3D super-resolution optical fluctuation imaging (SOFI).

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          Abstract

          Super-resolution optical microscopy is a rapidly evolving area of fluorescence microscopy with a tremendous potential for impacting many fields of science. Several super-resolution methods have been developed over the last decade, all capable of overcoming the fundamental diffraction limit of light. We present here an approach for obtaining subdiffraction limit optical resolution in all three dimensions. This method relies on higher-order statistical analysis of temporal fluctuations (caused by fluorescence blinking/intermittency) recorded in a sequence of images (movie). We demonstrate a 5-fold improvement in spatial resolution by using a conventional wide-field microscope. This resolution enhancement is achieved in iterative discrete steps, which in turn allows the evaluation of images at different resolution levels. Even at the lowest level of resolution enhancement, our method features significant background reduction and thus contrast enhancement and is demonstrated on quantum dot-labeled microtubules of fibroblast cells.

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          Author and article information

          Journal
          Proc Natl Acad Sci U S A
          Proceedings of the National Academy of Sciences of the United States of America
          Proceedings of the National Academy of Sciences
          1091-6490
          0027-8424
          Dec 29 2009
          : 106
          : 52
          Affiliations
          [1 ] Department of Chemistry and Biochemistry, and California NanoSystems Institute, University of California, Los Angeles, CA 90095, USA. t.dertinger@chem.ucla.edu
          Article
          0907866106
          10.1073/pnas.0907866106
          2799731
          20018714
          20d800e0-b318-402b-ad78-542c79b5c308
          History

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