Book excerpt: This volume focuses on various methods used by researchers to study mucosal-associated invariant T (MAIT) cells in the aspects described below. The chapters are organized into Four Parts: part One looks at the function and importance of MAIT cells in health and disease. It covers methods to isolate and characterize MAIT cells from human tissues including liver, colon tumors, placenta and decidua, and endometrium. Part Two discusses MR1 independent stimulation, and looks at the activation of MAIT cells by different stimulatory agents. Part Three describes various methods to develop tools to analyze, monitor, and generate MAIT cells in vitro, including the production of MR1-tetramers and how they can be loaded with bacterial antigens and used for the detection of MAIT cells. The last part covers the use of murine models to study MAIT cells. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and comprehensive, MAIT Cells: Methods and Protocols is a valuable resource for anyone interested in learning more about this developing field. Chapters 9 and 17 are open access under a CC BY 4.0 license.