Uncertainty in thematic maps has been tested mainly in maps with discrete or fuzzy classifications based on spectral data. However, many ecosystem maps in tropical countries consist of discrete polygons containing information on various ecosystem properties such as vegetation cover, soil, climate, geomorphology and biodiversity. The combination of these properties into one class leads to error. We propose a probability-based sampling design with two domains, multiple stages, and stratification with selection of primary sampling units (PSUs) proportional to the richness of strata present. Validation is undertaken through field visits and fine resolution remote sensing data. A pilot site in the center of the Colombian Andes was chosen to validate a government official ecosystem map. Twenty primary sampling units (PSUs) of 10 × 15 km were selected, and the final numbers of final sampling units (FSUs) were 76 for the terrestrial domain and 46 for the aquatic domain. Our results showed a confidence level of 95%, with the accuracy in the terrestrial domain varying between 51.8% and 64.3% and in the aquatic domain varying between 75% and 92%. Governments need to account for uncertainty since they rely on the quality of these maps to make decisions and guide policies.