Three-Dimensional Electron Microscopy of Macromolecular Assemblies

In this book, the mathematical principles and working methods of single-particle reconstruction are described; a method designed to retrieve three-dimensional structural information from electron micrographs showing thousands of “copies” of biological molecules trapped in a thin layer of ice. This technique is uniquely suited to obtain three-dimensional images of molecular machines in different functional states, as it dispenses with the need for crystals. The book starts with an introduction of image formation in the electron microscope, which includes the definition of the contrast transfer function. Next, averaging techniques and tools for image alignment, multivariate data analysis, and classification are…