Cryoem: Methods and Protocols

ISBN-13: 9781071609651 / Angielski / Twarda / 2020 / 350 str.

Tamir Gonen; Brent Nannenga

Spis treści

Part I – Electron Cryotomography

1. Getting Started with in situ cryo-electron Tomography

Daniel Serwas and Karen M. Davies

 

2. Cryo-Electron Tomography and Automatic Segmentation of Cultured Hippocampal Neurons

Ryan K. Hylton, Victoria H. Seader, and Matthew T. Swulius

 

3. Practical Approaches for Cryo-FIB Milling and applications for Cellular Cryo-Electron Tomography

Vinson Lam and Elizabeth Villa

 

4. In situ Imaging and Structure Determination of Biomolecular Complexes using Cryo-Electron Tomography

Mohammed Kaplan, William J Nicolas, Wei Zhao, Stephen D Carter, Lauren Ann Metskas, Georges Chreifi, Debnath Ghosal, and Grant J. Jensen

 

Part II– Single Particle Analysis and Electron Cryomicroscopy

5. Progress Towards CryoEM: Negative-stain Procedures for Biological Samples

Shane Gonen

 

6. Setting up Parallel Illumination on the Talos Arctica for High-Resolution Data Collection

Mark A. Herzik, Jr.

 

7. Multi-body refinement of cryo-EM images in RELION

Takanori Nakane and Sjors H.W. Scheres

 

8. Evaluating Local and Directional Resolution of Cryo-EM Density Maps

Sriram Aiyer, Cheng Zhang, Philp R. Baldwin, and Dmitry Lyumkis

 

9. Automated modeling and Validation of Protein Complexes in cryo-EM Maps

Tristan Cragnolini, Aaron Sweeney, and Maya Topf

 

10. 2D Electron Crystallography of Membrane Protein Single-, Double- and Multi-Layered Ordered Arrays

Matthew C. Johnson, Yusuf M. Uddin, Kasahun Neselu, and Ingeborg Schmidt-Krey

 

 

Part III–Electron Crystallography of 2d Crystals

11. Sample Preparation and Data Collection for Electron Crystallographic Studies on Membrane Protein Structures and Lipid-Protein Interaction

Ka-Yi Chan, Chloe Du Truong, Yu-Ping Poh, and Po-Lin Chiu

 

12. Single Particle Analysis for High Resolution 2D Electron Crystallography

Ricardo Righetto and Henning Stahlberg

 

13. MicroED Sample Preparation and Data Collection for Protein Crystals

Guanhong Bu and Brent L. Nannenga

 

PART IV – MICROCRYSTAL ELECTRON DIFFRACTION (MICROED)

14. Detection of Microcrystals for CryoEM

Simon Weiss, Sandra Vergara, Guowu Lin, and Guillermo Calero

 

15. Low-dose Data Collection and Radiation Damage in MicroED

Johan Hattne

 

16. Automation of Continuous-rotation Data Collection for MicroED

M. Jason de la Cruz

 

17. Ab initio Determination of Peptide Structures by MicroED

Chih-Te Zee, Ambarneil Saha, Michael R. Sawaya, and Jose A. Rodriguez

 

Blurb

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.