The book "Dictyostelium discoideum Protocols" offers a comprehensive collection of methods and techniques for studying the eukaryotic model organism Dictyostelium discoideum. This organism is known for its unique advantages in researching fundamental cellular processes such as signal transduction, cell motility, cytokinesis, and vesicle transport. The second edition of the book reflects the latest developments in various research areas and is divided into four parts. The first part provides an introduction to Amoebozoa and the available community resources. The second part addresses large-scale analytical methods made possible by the completion of the genome sequence of Dictyostelium. The third part focuses on molecular genetic techniques as well as cell biological, biochemical, and biophysical methods. The fourth part describes Dictyostelium as a model system for studying vesicle formation, transport, and infection by bacterial pathogens. The chapters are written in the established format of the "Methods in Molecular Biology" series and provide thorough introductions to their respective topics.