Interpolation of stream habitat can be very useful for habitat assessment. Using a small number of habitat samples to predict the habitat of larger areas can reduce time and labor costs as long as it provides accurate estimates of habitat. The spatial correlation of stream habitat variables such as substrate and depth improves the accuracy of interpolated data. Several geographical information system interpolation methods (natural neighbor, inverse distance weighted, ordinary kriging, spline, and universal kriging) were used to predict substrate and depth within a 210.7-m² section of a second-order stream based on 2.5% and 5.0% sampling of the total area. Depth and substrate were recorded for the entire study site and compared with the interpolated values to determine the accuracy of the predictions. In all instances, the 5% interpolations were more accurate for both depth and substrate than the 2.5% interpolations, which achieved accuracies up to 95% and 92%, respectively. Interpolations of depth based on 2.5% sampling attained accuracies of 49–92%, whereas those based on 5% percent sampling attained accuracies of 57–95%. Natural neighbor interpolation was more accurate than that using the inverse distance weighted, ordinary kriging, spline, and universal kriging approaches. Our findings demonstrate the effective use of minimal amounts of small-scale data for the interpolation of habitat over large areas of a stream channel. Use of this method will provide time and cost savings in the assessment of large sections of rivers as well as functional maps to aid the habitat-based management of aquatic species.