Subsurface flow in streambeds can vary at different scales in time and space. Recognizing this variability is critical for understanding biogeochemical and ecological processes associated with the hyporheic zone. The aim of this study was to examine the variability of hydraulic conductivity (K), vertical hydraulic gradients (VHGs), and subsurface fluxes, over a riffle-step-pool sequence and at a high spatio-temporal resolution. A 20 m reach was equipped with a network of piezometers in order to determine the distribution of VHGs and K. During a summer month, temporal variations of VHGs were regularly surveyed and, for a subset of piezometers, the water level was automatically recorded at 15 min intervals by logging pressure transducers. Additionally, point-dilution tests were carried out on the same subset of piezometers. Whereas the distribution of vertical fluxes can be derived from K and VHG values, point-dilution tests allow for the estimation of horizontal fluxes where no VHG is detectable. Results indicate that, spatially, VHGs switched from upwelling to downwelling across lateral as well as longitudinal sections of the channel. Vertical fluxes appeared spatially more homogeneous than VHGs, suggesting that the latter can be a poor indicator of the intensity of flow. Finally, during flow events, some VHGs showed little or no fluctuations; this was interpreted as the result of a pressure wave propagating from upstream through highly diffusive alluvial sediments.