Project description
Electron microscopy for 4D-STEM imaging in structural biology
Electron Microscopy (EM) has revolutionised both Life and Physical Sciences. Cryo-EM enables the determination of 3D protein structures with a resolution as fine as 1.0 Å. However, insufficient contrast impedes the determination of structures of molecules involved in diseases. EM offers optimal contrast for light atoms, achieving a remarkable resolution of 0.2 Å by transforming STEM from a 2D to a 4D imaging technique, effectively combining imaging with diffraction. The ERC-funded 4D-BioSTEM project aims to develop methodologies and a cryo-EM tool to enhance contrast and resolution in 4D-STEM imaging of frozen biological specimens. It uses specialised hardware, theory, simulation, and advanced image reconstruction algorithms. The project will acquire sparse 4D data and implement advanced techniques to extract maximum signal from noisy 4D cryo-STEM data.
Objective
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.
Fields of science
Keywords
Programme(s)
- HORIZON.1.1 - European Research Council (ERC) Main Programme
Topic(s)
Funding Scheme
HORIZON-ERC-SYG - HORIZON ERC Synergy GrantsHost institution
52428 Julich
Germany