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This volume details the most up-to-date cryo-EM techniques from leading researchers. Chapters are organized into four parts with emphasis on electron cryotomography, single particle analysis, and the crystal based cryo-EM methods of 2D electron crystallography, and MicroED for the study of 3D crystals. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, application details for both the expert and non-expert reader, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, CryoEM: Methods and Protocols aims to serve as an excellent resource on cryo-EM and can serve as the foundation for new researchers to this growing field in structural biology.
List of contents
Getting Started with in situ cryo-electron Tomography.- Cryo-Electron Tomography and Automatic Segmentation of Cultured Hippocampal Neurons.- Practical Approaches for Cryo-FIB Milling and applications for Cellular Cryo-Electron Tomography.- In situ Imaging and Structure Determination of Biomolecular Complexes using Cryo-Electron Tomography.- Progress Towards CryoEM: Negative-stain Procedures for Biological Samples.- Setting up Parallel Illumination on the Talos Arctica for High-Resolution Data Collection.- Multi-body refinement of cryo-EM images in RELION.- Evaluating Local and Directional Resolution of Cryo-EM Density Maps.- Automated modeling and Validation of Protein Complexes in cryo-EM Maps.- 2D Electron Crystallography of Membrane Protein Single-, Double- and Multi-Layered Ordered Arrays.- Sample Preparation and Data Collection for Electron Crystallographic Studies on Membrane Protein Structures and Lipid-ProteinInteraction.- Single Particle Analysis for High Resolution 2D Electron Crystallography.- MicroED Sample Preparation and Data Collection for Protein Crystals.-- Detection of Microcrystals for CryoEM.- Low-dose Data Collection and Radiation Damage in MicroED.- Automation of Continuous-rotation Data Collection for MicroED.- Ab initio Determination of Peptide Structures by MicroED.
