Live cell immunogold labelling of RNA polymerase II

Igor Orlov (1) , Andreas Schertel (2) , Guy Zuber (3) , Bruno Klaholz (1) , Robert Drillien (1) , Etienne Weiss (4) , Patrick Schultz (1) , Danièle Spehner (1)

1 IGBMC - Institut de Génétique et de Biologie Moléculaire et Cellulaire
2 Carl Zeiss Microscopy GmbH
3 CAMB - Conception et application de molécules bioactives
4 BSC - Biotechnologie et signalisation cellulaire

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.

Domaines: Organisation et fonctions cellulaires [q-bio.SC]

Igor Orlov, Andreas Schertel, Guy Zuber, Bruno Klaholz, Robert Drillien, et al.. Live cell immunogold labelling of RNA polymerase II. Scientific Reports, 2015, 5 (1), pp.8324. ⟨10.1038/srep08324⟩. ⟨hal-03535681⟩