Description du projet
La microscopie électronique pour l’imagerie 4D-STEM en biologie structurale
La microscopie électronique (ME) a révolutionné tant les sciences de la vie que les sciences physiques. La cryo-ME permet de caractériser les structures 3D des protéines avec une résolution aussi fine que 1,0 Å. Un contraste insuffisant empêche toutefois la détermination des structures des molécules impliquées dans les maladies. La ME offre un contraste optimal pour les atomes de la lumière, atteignant une résolution remarquable de 0,2 Å en transformant la STEM d’une technique d’imagerie 2D en une technique d’imagerie 4D, combinant efficacement l’imagerie et la diffraction. Le projet 4D-BioSTEM financé par le CER entend développer des méthodologies et un outil cryo-ME afin d’améliorer le contraste et la résolution dans l’imagerie 4D-STEM de spécimens biologiques congelés. Il fait appel à du matériel spécialisé, à la théorie, à la simulation et à des algorithmes avancés de reconstruction d’images. Le projet permettra d’acquérir des données 4D éparses et de mettre en œuvre des techniques avancées pour extraire le maximum de signal à partir de données cryo-STEM 4D bruyantes.
Objectif
Electron Microscopy (EM) has transformed research in the Life and Physical Sciences separately. Cryo-EM in Life Sciences allows the 3D structure determination of proteins down to 1.0 Å resolution in case they are large enough and present in high numbers in homogeneous states. For many molecules involved in diseases like neurodegeneration, however, structure determination is still severely hampered due to their insufficient contrast when imaged in vitreous ice or in their native cellular environment. To date, EM in the Physical Sciences generated utmost contrast for light atoms and established a resolution in the range of 0.2 Å limited only by thermal motion. This record was achieved by evolving scanning transmission EM (STEM) from a 2D to a 4D imaging technique combining imaging with diffraction. In this project, we will work out 4D-BioSTEM methodologies and develop a cryo-EM tool that maximizes contrast and resolution by bringing together EM groups from Life and Physical Sciences. 4D-STEM imaging of frozen biological specimens will be approached with unique and specialized hardware, theory and simulation, the development of microscope operation routines, and image reconstruction algorithms. In particular, we will acquire sparse 4D data using ultrafast detectors, employ methods for direct phasing (differential phase contrast, DPC) and establish advanced, so-called ptychographic, techniques to gain maximal signal from noisy 4D cryo-STEM data that are limited in their electron dose budget. We will make use of synergies between recently separated fields in order to make proteins smaller than 50 kDa amenable to structure elucidation. Furthermore, we will expand 4D-STEM to tomography, to obtain high-contrast 3D reconstructions from cellular samples, human brain tissue of neurodegenerative diseases, and vitrified organic energy materials. A new comprehensive structural imaging framework will be put forward and benchmarked as to its utility in Life and Physical Science applications of cryo-EM.
Champ scientifique
Mots‑clés
Programme(s)
- HORIZON.1.1 - European Research Council (ERC) Main Programme
Thème(s)
Régime de financement
HORIZON-ERC-SYG - HORIZON ERC Synergy GrantsInstitution d’accueil
52428 Julich
Allemagne