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Cryosectioning-enabled super-resolution microscopy for studying nuclear architecture at the single protein level.
Stein, Johannes; Ericsson, Maria; Nofal, Michel; Magni, Lorenzo; Aufmkolk, Sarah; McMillan, Ryan B; Breimann, Laura; Herlihy, Conor P; Lee, S Dean; Willemin, Andréa; Wohlmann, Jens; Arguedas-Jimenez, Laura; Yin, Peng; Pombo, Ana; Church, George M; Wu, Chao-Kng.
Afiliación
  • Stein J; Wyss Institute of Biologically Inspired Engineering, Boston, MA, USA.
  • Ericsson M; Department of Genetics, Harvard Medical School, Boston, MA, USA.
  • Nofal M; Blavatnik Institute, Harvard Medical School, Boston, MA, USA.
  • Magni L; Wyss Institute of Biologically Inspired Engineering, Boston, MA, USA.
  • Aufmkolk S; Wyss Institute of Biologically Inspired Engineering, Boston, MA, USA.
  • McMillan RB; Department of Genetics, Harvard Medical School, Boston, MA, USA.
  • Breimann L; Wyss Institute of Biologically Inspired Engineering, Boston, MA, USA.
  • Herlihy CP; Department of Genetics, Harvard Medical School, Boston, MA, USA.
  • Lee SD; Department of Genetics, Harvard Medical School, Boston, MA, USA.
  • Willemin A; Department of Genetics, Harvard Medical School, Boston, MA, USA.
  • Wohlmann J; Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetic Regulation and Chromatin Architecture Group, Berlin, Germany.
  • Arguedas-Jimenez L; Humboldt-Universität zu Berlin, Institute for Biology, Berlin, Germany.
  • Yin P; Department of Biosciences, University of Oslo, Norway.
  • Pombo A; Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetic Regulation and Chromatin Architecture Group, Berlin, Germany.
  • Church GM; Wyss Institute of Biologically Inspired Engineering, Boston, MA, USA.
  • Wu CK; Max-Delbrück-Center for Molecular Medicine in the Helmholtz Association (MDC), Berlin Institute for Medical Systems Biology (BIMSB), Epigenetic Regulation and Chromatin Architecture Group, Berlin, Germany.
bioRxiv ; 2024 Feb 05.
Article en En | MEDLINE | ID: mdl-38370628
ABSTRACT
DNA-PAINT combined with total Internal Reflection Fluorescence (TIRF) microscopy enables the highest localization precisions, down to single nanometers in thin biological samples, due to TIRF's unique method for optical sectioning and attaining high contrast. However, most cellular targets elude the accessible TIRF range close to the cover glass and thus require alternative imaging conditions, affecting resolution and image quality. Here, we address this limitation by applying ultrathin physical cryosectioning in combination with DNA-PAINT. With "tomographic & kinetically-enhanced" DNA-PAINT (tokPAINT), we demonstrate the imaging of nuclear proteins with sub-3 nanometer localization precision, advancing the quantitative study of nuclear organization within fixed cells and mouse tissues at the level of single antibodies. We believe that ultrathin sectioning combined with the versatility and multiplexing capabilities of DNA-PAINT will be a powerful addition to the toolbox of quantitative DNA-based super-resolution microscopy in intracellular structural analyses of proteins, RNA and DNA in situ.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: BioRxiv Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: BioRxiv Año: 2024 Tipo del documento: Article País de afiliación: Estados Unidos Pais de publicación: Estados Unidos