We describe a variety of one- and two-dimensional mathematical modeling approaches to characterizing diffuse flow circulation at mid-ocean ridge hydrothermal systems. The goal is to estimate the potential extent of the sub-seafloor microbial biosphere based on subsurface contours of the 120 °C isotherm as determined from the various models. The models suggest that the sub-seafloor depth for microbial life may range from less than 1 m in some places to the thickness of crustal layer 2A of in others. This depth depends primarily on how diffuse flow is driven. The 120 °C isotherm tends to be much deeper if diffuse flow is induced as boundary layer flow near high-temperature plumes, than if it results from conductive cooling or mixing near the seafloor. Because the heat flow alone may not allow identification of the flow regime in the subsurface, we highlight the use of chemical tracers as an additional constraint that sheds light into the flow and reaction patterns associated with vents. We use thermodynamic modeling, which connects the temperature of the diffuse fluid to its chemical composition. As the temperature-composition relationships differ for mixing versus conductive heating and cooling, the fluid geochemistry can shed light on subsurface transport. Using methane as an example, the geochemical models indicate subsurface microbial methane production and consumption in different regions of the vent field near EPR .