Workflow 2: Single-particle analysis cryoEM
Single-particle analysis (SPA) by cryoEM can achieve high resolution structural information of single proteins or protein complexes. Many identical copies of the particle of interest are embedded in vitreous ice on an EM grid by rapid plunge-freezing, or jet-freezing. Using automated data collection, thousands of images are collected, each containing hundreds of projections of the particles in various orientations. Computationally, these particle images are extracted, aligned and averaged, to reconstruct a high-resolution 3D map. This map is then used to build an atomic model of the protein structure.
CryoEM grid quality is typically assessed with our Glacios cryoTEM microscope, equipped with Falcon 3 direct electron detector. Automated SPA data collection is performed on one of our 3 Titan Krios microscopes, equipped with energy filters and K3, Falcon 3 or Falcon 4 direct electron detectors.