Groundwater flow advects heat, and thus, the deviation of subsurface temperatures from an expected conduction-dominated regime can be analysed to estimate vertical water fluxes. A number of analytical approaches have been proposed for using heat as a groundwater tracer, and these have typically assumed a homogeneous medium. However, heterogeneous thermal properties are ubiquitous in subsurface environments, both at the scale of geologic strata and at finer scales in streambeds. Herein, we apply the analytical solution of Shan and Bodvarsson (2004), developed for estimating vertical water fluxes in layered systems, in 2 new environments distinct from previous vadose zone applications. The utility of the solution for studying groundwater-surface water exchange is demonstrated using temperature data collected from an upwelling streambed with sediment layers, and a simple sensitivity analysis using these data indicates the solution is relatively robust. Also, a deeper temperature profile recorded in a borehole in South Australia is analysed to estimate deeper water fluxes. The analytical solution is able to match observed thermal gradients, including the change in slope at sediment interfaces. Results indicate that not accounting for layering can yield errors in the magnitude and even direction of the inferred Darcy fluxes. A simple automated spreadsheet tool (Flux-LM) is presented to allow users to input temperature and layer data and solve the inverse problem to estimate groundwater flux rates from shallow (e.g., <1 m) or deep (e.g., up to 100 m) profiles. The solution is not transient, and thus, it should be cautiously applied where diel signals propagate or in deeper zones where multi-decadal surface signals have disturbed subsurface thermal regimes.