Description du projet
Imagerie en temps réel de la différenciation des cellules souches
La microscopie optique est couramment utilisée pour la recherche et le diagnostic dans le monde entier, et permet de visualiser des objets d’une taille inférieure à 250 nm. La microscopie à super-résolution (SR) gagne du terrain sur la microscopie optique car elle contourne la limite de diffraction et améliore considérablement la capacité de résolution. Financé par le Conseil européen de l’innovation, le projet RT-SuperES vise à développer une technologie de SR automatisée qui offre la possibilité de réaliser une imagerie en temps réel pouvant passer de la microscopie conventionnelle à la microscopie de fluorescence SR. Les chercheurs prévoient d’utiliser ce système SR pour étudier la différenciation des cellules souches embryonnaires.
Objectif
The development of super-resolution (SR) microscopy in recent years has revolutionized cell biology, breaking the diffraction limit of light microscopy by order of magnitude. However, SR is currently incompatible with high-content imaging. RT-SuperES will provide a groundbreaking and affordable technology with automated SR capabilities beyond the state-of-the-art. To this end, we will generate a library of endogenously-labelled SNAP-tag fusion proteins in mouse embryonic stem cells (ESCs), and deploy a real-time decision-making module, which will continuously monitor our SNAP-tagged cells using fast fluorescence imaging, and, once a change is detected, will fix the desired cells, and switch to SR mode. By bringing together seven world-leading experts from four different countries, combining basic and applied research and industry, we propose several firsts: a) The first endogenously-labelled clone library of SNAP-tag fusion proteins; b) Utilize machine learning (ML) for real-time automated decision making, autonomously switching from fast conventional to SR imaging; c) Combine high content with SR imaging; d) Integrate novel, cutting-edge technologies, namely SR Radial Fluctuations (SRRF), NanoJ-Fluidics, Single Molecule Localization Microscopy (SMLM) and Structured Illumination Microscopy (SIM); e) Collect large scale imaging datasets of cell states in ESCs, and f) Provide cell-cycle stage-dependent nanoscale localization of selected nuclear and chromatin proteins (e.g. H3.3) during early ESC differentiation. RT-SuperES will provide the scientific community with the first-of-its-kind commercial real-time SR-highcontent imaging system, and the first library of endogenously SNAP-tagged ESC clones, which are ideal, among many other things, for SR imaging.
Champ scientifique
- natural sciencesphysical sciencesopticsmicroscopysuper resolution microscopy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesbiological sciencescell biology
- medical and health sciencesmedical biotechnologycells technologiesstem cells
- natural sciencescomputer and information sciencesartificial intelligencemachine learning
Mots‑clés
Programme(s)
- HORIZON.3.1 - The European Innovation Council (EIC) Main Programme
Régime de financement
EIC - EICCoordinateur
91904 Jerusalem
Israël