Ambient ionization mass spectrometry allows the rapid analysis of samples or objects in their native state in the open environment with no prior preparation. Over the past six years, the ability of these techniques to provide selective analyte desorption and ionization, in combination with mass spectrometry (MS), has provided a growing number of powerful analytical alternatives across broad application areas, both quantitative and qualitative in nature, including pharmaceutical analysis, process chemistry, biological imaging, in vivo analysis, proteomics, metabolomics, forensics, and explosives detection. With the emergence of new ambient ionization methods, and the complementary nature of existing desorption and/or ionization techniques, additional hyphenated methods have been devised, which pushes the total number of documented methods to almost thirty. To cover all current ambient ionization techniques in detail would be too complex and detract from the main objective of this review. Rather, an overview of the field of ambient ionization MS will be given, followed by broad classification to allow detailed discussion of theory and common mechanistic factors underpinning a number of key techniques. Consideration will be given to experimental design, ease of implementation and analytical performance, detailing subsequent impact on a number of application areas, both established and emerging.