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      Imaging intracellular fluorescent proteins at nanometer resolution.

      Science (New York, N.Y.)

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