The Microsetta Initiative (TMI) consolidates our global efforts to collect microbiome samples of the world’s populations and to couple these collections with educational outreach about microbiome science. TMI is part of the Earth Microbiome Project, and the data produced are de-identified and placed into the public domain so that scientists around the world contribute to research in understanding how health and lifestyle factors associated with the microbiome translate between populations. TMI was conceived as the next natural step on the evolution of the American Gut Project, to increase its geographical reach, and provide a framework for its international expansion efforts. TMI is a highly collaborative effort, with an emphasis on inclusion, standardization of technical practices, and utilization of modern molecular methodologies. This transition emphasizes engagement with underrepresented populations both at the local level and global scale and expansion of the availability of population-specific microbiome references to improve the interpretation of regional studies.