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Sci Rep. 2015 Feb 9;5:8324. doi: 10.1038/srep08324.

Live cell immunogold labelling of RNA polymerase II.

Author information

1
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.
2
Carl Zeiss Microscopy GmbH, Training, Application and Support Center (TASC), Carl-Zeiss-Straße 22, D-73447 Oberkochen, Germany.
3
LCAMB, UMR 7199, CNRS/Strasbourg University, Faculty of Pharmacy, 74 route du Rhin, 67401 Illkirch, France.
4
LBSC, UMR 7242, Institut de Recherche de l'Ecole de Biotechnologie de Strasbourg, Strasbourg University, 300 Boulevard Sébastien Brandt, BP 10413, 67412 Illkirch, France.

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.

PMID:
25662860
PMCID:
PMC4321181
DOI:
10.1038/srep08324
[Indexed for MEDLINE]
Free PMC Article

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