Abstract
Recent improvements in direct electron detectors, microscope technology and software provided the stimulus for a `quantum leap' in the application of cryo-electron microscopy in structural biology, and many national and international centres have since been created in order to exploit this. Here, a new facility for cryo-electron microscopy focused on single-particle reconstruction of biological macromolecules that has been commissioned at the European Synchrotron Radiation Facility (ESRF) is presented. The facility is operated by a consortium of institutes co-located on the European Photon and Neutron Campus and is managed in a similar fashion to a synchrotron X-ray beamline. It has been open to the ESRF structural biology user community since November 2017 and will remain open during the 2019 ESRF-EBS shutdown.
Highlights
Recent progress in direct electron detector technology, the provision of intense and coherent electron beams using field emission guns (FEGs and X-FEGs) in cryo-electron microscopes and advances in image-processing algorithms and sample preparation have led to the so-called ‘resolution revolution’ (Kuhlbrandt, 2014; Mitra, 2019) in cryo-electron microscopy
In 2015, the European Synchrotron Radiation Facility (ESRF) took the decision to complement its portfolio of cutting-edge X-ray-based facilities for structural biology (Pernot et al, 2013; von Stetten et al, 2015; Linden et al, 2014; Mueller-Dieckmann et al, 2015) by acquiring, installing and commissioning a cryo-electron microscopy (cryo-EM) facility (CM01; http://www.esrf.eu/ home/UsersAndScience/Experiments/MX/About_our_beamlines/ CM01.html) based around a Titan Krios microscope and making it available to its structural biology user community
Preliminary sample screening will be carried out using the Partnership for Structural Biology (PSB) cryo-EM platform microscopes located at the Grenoble Institut de Biologie Structurale (IBS)
Summary
Recent progress in direct electron detector technology, the provision of intense and coherent electron beams using field emission guns (FEGs and X-FEGs) in cryo-electron microscopes and advances in image-processing algorithms and sample preparation have led to the so-called ‘resolution revolution’ (Kuhlbrandt, 2014; Mitra, 2019) in cryo-electron microscopy (cryo-EM). An average resolution for singleparticle cryo-EM structures of between 5 and 6 Afor the 3DEM density maps deposited suggests that there is still some way to go before cryo-EM becomes a routine technique for the production of high-resolution structural information With this aim in mind, and coupled with significant advances in sample preparation and data processing, many national and international centres for cryo-EM have opened during the last few years (see, for example, Alewijnse et al, 2017, and references therein; Stuart et al, 2016; Clare et al, 2017). Preliminary sample screening will be carried out using the PSB cryo-EM platform (http:// www.ibs.fr/research/research-groups/ methods-and-electron-microscopy-group/ electron-microscopy-platform/) microscopes located at the Grenoble Institut de Biologie Structurale (IBS) Once this mode of operation is fully functional, it will provide a unique opportunity for scientists of the ESRF’s international community with limited or no access to EM facilities to access the technique for scientifically important projects
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