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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.

SUBMITTER: Orlov I

PROVIDER: S-EPMC4321181 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature

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Live cell immunogold labelling of RNA polymerase II.

Orlov Igor I Schertel Andreas A Zuber Guy G Klaholz Bruno B Drillien Robert R Weiss Etienne E Schultz Patrick P Spehner Danièle D

Scientific reports 20150209

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. ...[more]

PMID: 25662860

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