Live cell immunogold labelling of RNA polymerase II

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / metabolism*
  • Gold*
  • HeLa Cells
  • Humans
  • Microscopy, Electron* / methods
  • RNA Polymerase II / metabolism*
  • Staining and Labeling* / methods

Substances

  • Gold
  • RNA Polymerase II