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The Atacama Desert, one of the most arid landscapes on Earth, serves as an analog for the dry conditions on Mars and as a test bed in the search for life on other planets. During the Life in the Atacama (LITA) 2004 field experiment, satellite imagery and ground-based rover data were used in concert with a `follow-the-water’ exploration strategy to target regions of biological interest in two ( 1 coastal, 1 inland) desert study sites. Within these regions, environments were located, studied and mapped with spectroscopic and fluorescence imaging (FI) for habitats and microbial life. Habitats included aqueous sedimentary deposits ( e. g., evaporites), igneous materials ( e. g., basalt, ash deposits), rock outcrops, drainage channels and basins, and alluvial fans. Positive biological signatures ( chlorophyll, DNA, protein) were detected at 81% of the 21 locales surveyed with the FI during the long-range, autonomous traverses totaling 30 km. FI sensitivity in detecting microbial life in extreme deserts explains the high percentage of positives despite the low actual abundance of heterotrophic soil bacteria in coastal (< 1-10(4) CFU/g-soil) and interior (< 1-10(2) CFU/g-soil) desert soils. Remote habitat, microbial and climate observations agreed well with ground-truth, indicating a drier and less microbially rich interior compared to the relatively wetter and abundant biology of the coastal site where rover sensors detected the presence of fog and abundant surface lichens. LITA project results underscore the importance of an explicit focus by all engineering and science disciplines on microbially relevant scales ( mm to nm), and highlight the success of satellite-based and `follow-the-water’ strategies for locating diverse habitats of biological promise and detecting the microbial hotspots within them.