Conventional scanning and transmission electron microscopy (SEM and TEM) of the rhizosphere (soil around plant roots) has been used to describe the spatial organization of biotic (microorganisms including bacteria and fungi) and abiotic (organomineral complexes and clay platelets) constituents in soil.1 However, soil is deeper than plant rootS2 and the spatial distribution of microbes, the morphology of the microbial growth habit, and the spatial association of microbes with mineral substrates in the deeper vadose zone are little known. That subsurface terrestrial microbes catalyze most major nutrient-cycling reactions in the terrestrial biosphere3 punctuates the need to observe the true morphology of bacteria in association with mineral surfaces and microbemicrobe associations. Here, we present our use of environmental scanning electron microscopy (ESEM) to image bacteria on unsaturated sand, a simple surrogate for heterogeneous soil. ESEM is a choice tool for high resolution imaging of combined soft and hard matter, where delicate hydrated expolymer matrices bind microbes to each other and to soil particles as biofilm.4