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      Imaging intracellular fluorescent proteins at nanometer resolution.

      Science (New York, N.Y.)

      methods, Animals, Fluorescence, COS Cells, Photobleaching, Algorithms, Mitochondria, Recombinant Fusion Proteins, analysis, Gene Products, gag, HIV-1, Nanotechnology, Pseudopodia, Luminescent Proteins, chemistry, Microscopy, Cercopithecus aethiops, Actins, Cell Membrane, Proteins, Light, Focal Adhesions, Organelles, Lysosomes, Cell Line, Vinculin

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          Abstract

          We introduce a method for optically imaging intracellular proteins at nanometer spatial resolution. Numerous sparse subsets of photoactivatable fluorescent protein molecules were activated, localized (to approximately 2 to 25 nanometers), and then bleached. The aggregate position information from all subsets was then assembled into a superresolution image. We used this method--termed photoactivated localization microscopy--to image specific target proteins in thin sections of lysosomes and mitochondria; in fixed whole cells, we imaged vinculin at focal adhesions, actin within a lamellipodium, and the distribution of the retroviral protein Gag at the plasma membrane.

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          Journal
          16902090
          10.1126/science.1127344

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