This comprehensive book gives a systematic introduction to the techniques of single-particle cryo-EM of macromolecular machines. The first book of its kind, it focuses on not only the digital image reconstruction procedures, but includes an introductory chapter on the theory of image formation, specimen preparation, data collection, sources of noise. One whole chapter is dedicated to the practical procedures for structure determination. The methods are described from the authors' practical point of view. Developments, such as parallel computing, automated docking, and field emission gun (FEG) cryo-EM, described illustrate recent successes in the field in the solution of structures. Sufficient detail of the theory of digital imaging processing allows the interested biologist to understand the underlying mathematical and computational concepts of the method. Ideal not only for experienced scientist, postdocs, graduate students working in the field of cryo-EM, but also students of biophysics and structural biology. Newcomers benefit greatly from this treatment of the subject.