title: Imaging intracellular fluorescent proteins at nanometer resolution.
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.
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA. betzige@janelia.hhmi.org
authors.forename: Eric
authors.initials: E
authors.lastname: Betzig
authors.forename: George H
authors.initials: GH
authors.lastname: Patterson
authors.forename: Rachid
authors.initials: R
authors.lastname: Sougrat
authors.forename: O Wolf
authors.initials: OW
authors.lastname: Lindwasser
authors.forename: Scott
authors.initials: S
authors.lastname: Olenych
authors.forename: Juan S
authors.initials: JS
authors.lastname: Bonifacino
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lippincott-Schwartz
authors.forename: Harald F
authors.initials: HF
authors.lastname: Hess
mesh_terms: D000199:Actins / Q000032:analysis; D000465:Algorithms; D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry*; D002522:Chlorocebus aethiops; D005453:Fluorescence; D022001:Focal Adhesions / Q000737:chemistry; D015683:Gene Products, gag / Q000032:analysis; D015497:HIV-1; D008027:Light; D008164:Luminescent Proteins / Q000032:analysis*; D008247:Lysosomes / Q000737:chemistry; D008853:Microscopy / Q000379:methods*; D008928:Mitochondria / Q000737:chemistry; D036103:Nanotechnology; D015388:Organelles / Q000737:chemistry*; D038761:Photobleaching; D011506:Proteins / Q000032:analysis*; D011554:Pseudopodia / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D016596:Vinculin / Q000032:analysis
chemical_list: D000199:Actins; D015683:Gene Products, gag; D008164:Luminescent Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins; D016596:Vinculin