title: MRT letter: Nanoscopy of protein colocalization in living cells by STED and GSDIM.
abstract: We report the use of superresolution fluorescence microscopy for studying the nanoscale distribution of protein colocalization in living mammalian cells. Nanoscale imaging is attained both by a targeted and a stochastic fluorescence on-off switching superresolution method, namely by stimulated emission depletion (STED) and ground state depletion microscopy followed by individual molecular return (GSDIM), respectively. Analysis of protein colocalization is performed by bimolecular fluorescence complementation (BiFC). Specifically, a nonfluorescent fragment of the yellow fluorescent protein Citrine is fused to tubulin while a counterpart nonfluorescent fragment is fused to the microtubulin-associated protein MAP2 such that fluorescence is reconstituted on contact of the fragment-carrying proteins. Images with resolution down to 65 nm prove a powerful new way for studying protein colocalization in living cells at the nanoscale.
authors.affiliation: Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, Göttingen, Germany.
authors.forename: Birka
authors.initials: B
authors.lastname: Lalkens
authors.forename: Ilaria
authors.initials: I
authors.lastname: Testa
authors.forename: Katrin I
authors.initials: KI
authors.lastname: Willig
authors.forename: Stefan W
authors.initials: SW
authors.lastname: Hell
mesh_terms: D000818:Animals; D002477:Cells / Q000166:cytology / Q000378:metabolism*; D002522:Chlorocebus aethiops; D008164:Luminescent Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D021381:Protein Transport; D011506:Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells
chemical_list: D008164:Luminescent Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins