Water-mediated linkages that connect landscape components are collectively referred to as hydrologic connectivity. These linkages influence numerous watershed processes including biogeochemical cycling, spatial vegetation patterns, and stream runoff generation. The concept of hydrologic connectivity also informs environmental management and underpins regulations protecting waterways. However, to date, there is no consensus on how to quantitatively assess connectivity. Here, we develop and test a framework to quantify hydrologic connectivity within a landscape. We define connectivity as a continuous variable (from 0 to 1) that represents the vector strength between any two points in the landscape (a source to a target). To measure this vector strength, we analyzed hydrologic and geochemical indicators within a montane river-floodplain system across a dynamic range of streamflows. In addition to applying hydrologic and geochemical indicators, we tested the ability of microbiome membership to provide further insight into connectivity dynamics. From these data, we generated complementary time series of lateral connectivity (between the river and the floodplain) and longitudinal connectivity (along the river from upstream to downstream). We then quantified key parameters associated with connectivity regimes among waterbodies including connectivity strength and stability, and timing and speed of changes in connectivity. The application of a microbial index for connectivity provided new insight into flowpath residence times that was not apparent using more traditional hydro-geochemical approaches. The proposed connectivity framework moves beyond binary qualitative descriptions of connectivity and provides a coupled conceptual and empirical approach to quantify the spatiotemporal variability of hydrologic connectivity.