Current land-management decisions that affect the persistence of native salmonids are often influenced by studies of individual sites that are selected based on judgment and convenience. Although this approach is useful for some purposes, extrapolating results to areas that were not sampled is statistically inappropriate because the sampling design is usually biased. Therefore, in recent investigations of coastal cutthroat trout (Oncorhynchus clarki clarki) located above natural barriers to anadromous salmonids, we used a methodology for extending the statistical scope of inference. The purpose of this paper is to apply geospatial tools to identify a population of watersheds and develop a probability-based sampling design for coastal cutthroat trout in western Oregon, USA. The population of mid-size watersheds (500-5800 ha) west of the Cascade Range divide was derived from watershed delineations based on digital elevation models. Because a database with locations of isolated populations of coastal cutthroat trout did not exist, a sampling frame of isolated watersheds containing cutthroat trout had to be developed. After the sampling frame of watersheds was established, isolated watersheds with coastal cutthroat trout were stratified by ecoregion and erosion potential based on dominant bedrock lithology (i.e., sedimentary and igneous). A stratified random sample of 60 watersheds was selected with proportional allocation in each stratum. By comparing watershed drainage areas of streams in the general population to those in the sampling frame and the resulting sample (n = 60), we were able to evaluate the how representative the subset of watersheds was in relation to the population of watersheds. Geospatial tools provided a relatively inexpensive means to generate the information necessary to develop a statistically robust, probability-based sampling design.