Brief Communication abstract


Nature Methods 5, 1027 - 1030 (2008)
Published online: 16 November 2008 | doi:10.1038/nmeth.1271

Nanoscale imaging of molecular positions and anisotropies

Travis J Gould1,5, Mudalige S Gunewardene1,5, Manasa V Gudheti1, Vladislav V Verkhusha2, Shu-Rong Yin3, Julie A Gosse4 & Samuel T Hess1

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Knowledge of the orientation of molecules within biological structures is crucial to understanding the mechanisms of cell function. We present a method to image simultaneously the positions and fluorescence anisotropies of large numbers of single molecules with nanometer lateral resolution within a sample. Based on a simple modification of fluorescence photoactivation localization microscopy (FPALM), polarization (P)-FPALM does not compromise speed or sensitivity. We show results for mouse fibroblasts expressing Dendra2-actin or Dendra2-hemagglutinin.

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  1. Department of Physics and Astronomy and Institute for Molecular Biophysics, 5709 Bennett Hall, University of Maine, Orono, Maine 04469, USA.
  2. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461, USA.
  3. Laboratory for Cellular and Molecular Biophysics, Program in Physical Biology, National Institute of Child Health and Human Development, 10 Center Drive, Bethesda, Maryland 20892, USA.
  4. Department of Biochemistry, Microbiology, and Molecular Biology, 5735 Hitchner Hall, University of Maine, Orono, Maine 04469, USA.
  5. These authors contributed equally to this work.

Correspondence to: Samuel T Hess1 e-mail: sam.hess@umit.maine.edu




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