Deep biosphere and subseafloor hydrosphere studies are critical focus areas for the Integrated Ocean Drilling Program (IODP). In this project, researchers will develop and test new instrumentation for the incubation of microbes in boreholes that may be drilled and CORKed at North Pond, on the western flank of the Mid-Atlantic Ridge at 22 degrees 45 minutes north, under the auspices of a pending IODP project. This project will also extend analysis techniques (Flourescently-Labeled Embedded Core or FLEC) that permit careful study of the spatial relationships ('life locations') among microbes for use in various microbial studies in the deep marine environment, including on the incubators to be developed in this project. FLEC, combined with established ribosomal RNA detection methods (CARD and FISH), will provide significantly more information about the microbial systems, including community structure and microbial associations at a fine scale. Broader impacts include training of graduate and undergraduate students, advancement of IODP science and building on ODP science, hosting of a workshop, and educational outreach to teacher groups.