Gold-labels

Functionalized gold labels to visualize biomolecules and ligands by electron microscopy



Résumé
Cryo-Electron Tomography (ET) offers the possibility to visualize the molecular organization of protein complexes within their native cellular environment. A current challenge is to identify key functional sites for transcription within the cell nucleus, and to determine their structure and molecular environment. Molecular complexes of low cellular abundance or too small to be recognized by their shape need to be labelled with an electron dense marker to be detected with good statistical significance. To address this challenge, we propose to synthesize novel gold-based probes that can be delivered into living cells. The gold particle will be coupled to antibody derivatives and we will optimize its properties to selectively label the target proteins. The gold particles will be detected in cell sections using STEM-HAADF to determine their spatial distribution. Once optimized RNA polymerase II molecules will be gold-labelled in living cells, the cell will be vitrified and thin lamellas will be produced by Focused Ion Beam milling before cryo-ET analysis.

Mots-clés
functionalized gold particles, cryo electron tomography, STEM-HAADF, transcription factories, surface functionalization

Partenaires du projet

INSB
SCHULTZ Patrick
(UMR7104) Illkirch-Graffenstaden France
INSB
Zuber Guy
Biotechnologie et Signalisation Cellulaire (UMR7242) France
INP
ERSEN Ovidiu
Institut de Physique et de Chimie des Matériaux de Strasbourg (IPCMS) (UMR7504) France
Patrick Schultz
Crédit photo : Patrick Schultz