Cryo-EM Part A: Sample Preparation and Data Collection

Cryo-EM Part A: Sample Preparation and Data Collection is dedicated to a description of the instruments, samples, protocols, and analyses that belong to cryo-EM. It emphasizes the relatedness of the ideas, instrumentation, and methods underlying all cryo-EM approaches, which allow practitioners to easily move between them. Within each section, the articles are ordered according to the most common symmetry of the sample to which their methods are applied.

Includes time-tested core methods and new innovations applicable to any researcherMethods included are useful to both established researchers and newcomers to the fieldRelevant background and reference information given for procedures can be used as a guide

Historical perspective

3D reconstruction from electron micrographs: a personal account of its development

David DeRosier

1. Preparation of 2-D crystals

Henning Stahlberg

2. Helical Crystallization of Soluble and Membrane Binding Proteins

Elizabeth M. Wilson-Kubalek, Christopher Arthur, Joshua S. Chappie

3. Plunge Freezing for Electron Cryomicroscopy

Alasdair W. McDowall, Megan J. Dobro, Linda A. Melanson, Grant J. Jensen

4. A Practical Guide to the Use of Monolayer Purification and Affinity Grids

Thomas Walz, Deborah F. Kelly, Danijela Dukovski

5. GraFix: Stabilization of fragile macromolecular complexes for single particle cryo-EM

Holger Stark

6. Cryo-negative staining of macromolecular assemblies

Sacha De Carlo, Holger Stark

7. Liposomes on a streptavidin crystal: a system to study membrane proteins by cryo-EM

Fred J. Sigworth, Liguo Wang

8. Micromanipulator-Assisted Vitreous Cryosectioning and Sample Preparation by High-Pressure Freezing

Mark S. Ladinsky

9. Site-Specific Biomolecule Labeling with Gold Clusters

Christopher J. Ackerson, Richard D. Powell, James F. Hainfeld

10. How to Operate a Cryo-Electron Microscope

Huilin Li, Jingchuan Sun

11. Collecting electron crystallographic data of two-dimensional protein crystals

Thomas Walz, Richard K. Hite, Andreas D. Schenk, Zongli Li, Yifan Cheng

12. Automated Data Collection for Electron Microscopic Tomography

David Agard, Shawn Q. Zheng, J. W. Sedat

13. Correlated Light and Electron Cryomicroscopy

Grant J. Jensen, Ariane Briegel, Songye Chen, Abraham J. Kosterc, Jürgen M. Plitzkod, Cindi L. Schwartz

14. Phase Plates for Transmission Electron Microscopy

Radostin Danev, Kuniaki Nagayama

15. Radiation damage in electron cryomicroscopy

John L. Rubinstein, Lindsay A. Baker