Spatially intensive sampling by electrofishing is proposed as a method for quantifying spatial variation in fish assemblages at multiple scales along extensive stream sections in headwater catchments. We used this method to sample fish species at 10-m2 points spaced every 20 m throughout 5 km of a headwater stream in France. The spatially intensive sampling design provided information at a spatial resolution and extent that enabled exploration of spatial heterogeneity in fish assemblage structure and aquatic habitat at multiple scales with empirical variograms and wavelet analysis. These analyses were effective for detecting scales of periodicity, trends, and discontinuities in the distribution of species in relation to tributary junctions and obstacles to fish movement. This approach to sampling riverine fishes may be useful in fisheries research and management for evaluating stream fish responses to natural and altered habitats and for identifying sites for potential restoration.