title: Live cell immunogold labelling of RNA polymerase II.
abstract: Labeling nuclear proteins with electron dense probes in living cells has been a major challenge due to their inability to penetrate into nuclei. We developed a lipid-based approach for delivering antibodies coupled to 0.8 nm ultrasmall gold particles into the nucleus to label RNA polymerase II. Focussed Ion Beam slicing coupled to Scanning Electron Microscopy (FIB/SEM) enabled visualization of entire cells with probe localization accuracy in the 10 nm range.
authors: Department of Integrated Structural Biology, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) INSERM, U964; CNRS/Strasbourg University, UMR7104 1, rue Laurent Fries, BP10142 67404 Illkirch, France.
authors: Igor
authors: I
authors: Orlov
authors: Carl Zeiss Microscopy GmbH, Training, Application and Support Center (TASC), Carl-Zeiss-Straße 22, D-73447 Oberkochen, Germany.
authors: Andreas
authors: A
authors: Schertel
authors: LCAMB, UMR 7199, CNRS/Strasbourg University, Faculty of Pharmacy, 74 route du Rhin, 67401 Illkirch, France.
authors: Guy
authors: G
authors: Zuber
authors: Department of Integrated Structural Biology, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) INSERM, U964; CNRS/Strasbourg University, UMR7104 1, rue Laurent Fries, BP10142 67404 Illkirch, France.
authors: Bruno
authors: B
authors: Klaholz
authors: Department of Integrated Structural Biology, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) INSERM, U964; CNRS/Strasbourg University, UMR7104 1, rue Laurent Fries, BP10142 67404 Illkirch, France.
authors: Robert
authors: R
authors: Drillien
authors: LBSC, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg, Strasbourg University, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.
authors: Etienne
authors: E
authors: Weiss
authors: Department of Integrated Structural Biology, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) INSERM, U964; CNRS/Strasbourg University, UMR7104 1, rue Laurent Fries, BP10142 67404 Illkirch, France.
authors: Patrick
authors: P
authors: Schultz
authors: Department of Integrated Structural Biology, IGBMC (Institut de Génétique et de Biologie Moléculaire et Cellulaire) INSERM, U964; CNRS/Strasbourg University, UMR7104 1, rue Laurent Fries, BP10142 67404 Illkirch, France.
authors: Danièle
authors: D
authors: Spehner
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism*; D006046:Gold; D006367:HeLa Cells; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods; D012319:RNA Polymerase II / Q000378:metabolism*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D006046:Gold; D012319:RNA Polymerase II