Given projected increases in stream temperatures attributable to global change, improved understanding of relationships between stream temperatures and hyporheic exchange would be useful. We conducted two conservative tracer injection experiments in a glacial meltwater stream, to evaluate the effects of hyporheic thermal gradients on exchange processes, including preferential flow paths (PFPs). The experiments were conducted on the same day, the first (a stream injection) during a cool, morning period and the second (dual stream and hyporheic injections) during a warm, afternoon period. In the morning, the hyporheic zone was thermally uniform at 4°C, whereas by the afternoon the upper 10 cm had warmed to 6–12°C and exhibited greater temperature heterogeneity. Solute transport modeling showed that hyporheic cross-sectional areas (As) at two downstream sites were two and seven times lower during the warm experiment. Exchange metrics indicated that the hyporheic zone had less influence on downstream solute transport during the warm, afternoon experiment. Calculated hyporheic depths were less than 5 cm, contrasting with tracer detection at 10 and 25 cm depths. The hyporheic tracer arrival at one downstream site was rapid, comparable to the in-stream tracer arrival, providing evidence for PFPs. We thus propose a conceptual view of the hyporheic zone in this reach as being dominated by discrete PFPs weaving through hydraulically isolated areas. One explanation for the simultaneous increase in temperature heterogeneity and As decrease in a warmer hyporheic zone may be a flow path preferentiality feedback mechanism resulting from a combination of temperature-related viscosity decreases and streambed heterogeneity.