Synopses & Reviews
Cryoelectron microscopy of biological molecules is among the hottest growth areas in biophysics and structural biology at present, and Frank is arguably the most distinguished practitioner of this art. CryoEM is likely over the next few years to take over much of the structural approaches currently requiring X-ray crystallography, because one can now get good and finely detailed images of single molecules down to as little as 200,000 MW, covering a substantial share of the molecules of greatest biomedical research interest. This book, the successor to an earlier work published in 1996 with Academic Press, is a natural companion work to our forthcoming book on electron crystallography by Robert Glaeser, with contributions by six others, including Frank. A growing number of workers will employ CryoEM for structural studies in their own research, and a large proportion of biomedical researchers will have a growing interest in understanding what the capabilities and limits of this approach are.
Synopsis
In the past ten years there has been a tremendous surge in applications of cryo-electron microscopy in the study of macromolecular assemblies, in especially those in single-particle form. Although the path toward atomic resolution is much more tedious than in electron crystallography of two-dimensional crystals, realization of such resolution for entirely asymmetrical molecules is now in close reach. In Three-Dimensional Electron Microscopy of Macromolecular Assemblies: Visualization of Biological Molecules in Their Native State, Joachim Frank describes the mathematical principles and working methods of single-particle reconstruction--a method designed to retrieve structural information from electron micrographs showing many -copies- of molecules trapped in ice in random orientations. The technique is uniquely suited to obtain three-dimensional images of molecular machines in different functional states, as it does not require crystals. This revised edition includes a basic introduction to transmission electron microscopy and the design of the instruments, and it covers specimen preparation in greater depth. Frank addresses the remarkable convergence of processing methods that has taken place, making it possible to sharpen the focus of some sections. New sections have been added that cover segmentation as well as various techniques of docking and fitting. Finally, appendix material is now included; this tabulates applications and methods of single-particle reconstruction and describes the most important mathematical tools and theorems of Fourier processing.
About the Author
Joachim Frank is an Investigator at the Howard Hughes Medical Institute. He is also Professor of Biomedical Sciences and Adjunct Professor of Biology at the State University of New York at Albany, Adjunct Professor of Biochemistry and Molecular Biophysics at Columbia University, Research Professor of Cell Biology at New York State University Medical School, and head of the Laboratory of Computational Biology and Macromolecular Imaging at Health Research, Inc., the Wadsworth Center.
Table of Contents
1. Introduction
2. Electron Microscopy of Macromolecular Assemblies
3. Two-Dimensional Averaging Techniques
4. Multivariate Data Analysis and Classification of Images
5. Three-Dimensional Reconstruction
6. Interpretation of Three-Dimensional Images of Macromolecules
Appendix 1: Some Important Definitions and Theorems
Appendix 2: Profiles, Point-Spread Functions, and Effects of Commonly Used Low-Pass Filters
Appendix 2: Bibliography of Methods
Appendix 2: Bibliography of Structures
Appendix 2: Special Journal Issues on Image Processing Techniques