title: Interferometric fluorescent super-resolution microscopy resolves 3D cellular ultrastructure.
abstract: Understanding molecular-scale architecture of cells requires determination of 3D locations of specific proteins with accuracy matching their nanometer-length scale. Existing electron and light microscopy techniques are limited either in molecular specificity or resolution. Here, we introduce interferometric photoactivated localization microscopy (iPALM), the combination of photoactivated localization microscopy with single-photon, simultaneous multiphase interferometry that provides sub-20-nm 3D protein localization with optimal molecular specificity. We demonstrate measurement of the 25-nm microtubule diameter, resolve the dorsal and ventral plasma membranes, and visualize the arrangement of integrin receptors within endoplasmic reticulum and adhesion complexes, 3D protein organization previously resolved only by electron microscopy. iPALM thus closes the gap between electron tomography and light microscopy, enabling both molecular specification and resolution of cellular nanoarchitecture.
authors.affiliation: Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, VA 20147, USA.
authors.forename: Gleb
authors.initials: G
authors.lastname: Shtengel
authors.forename: James A
authors.initials: JA
authors.lastname: Galbraith
authors.forename: Catherine G
authors.initials: CG
authors.lastname: Galbraith
authors.forename: Jennifer
authors.initials: J
authors.lastname: Lippincott-Schwartz
authors.forename: Jennifer M
authors.initials: JM
authors.lastname: Gillette
authors.forename: Suliana
authors.initials: S
authors.lastname: Manley
authors.forename: Rachid
authors.initials: R
authors.lastname: Sougrat
authors.forename: Clare M
authors.initials: CM
authors.lastname: Waterman
authors.forename: Pakorn
authors.initials: P
authors.lastname: Kanchanawong
authors.forename: Michael W
authors.initials: MW
authors.lastname: Davidson
authors.forename: Richard D
authors.initials: RD
authors.lastname: Fetter
authors.forename: Harald F
authors.initials: HF
authors.lastname: Hess
mesh_terms: D000818:Animals; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000295:instrumentation* / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation* / Q000379:methods*; D008857:Microscopy, Interference / Q000295:instrumentation* / Q000379:methods*; D008870:Microtubules