In the context of the already observed population changes at specific time intervals and the shift in penguin breeding sites because of global warming and the fluctuations in availability of food, the report presents the methodical tools for an area-wide penguin monitoring of Antarctic penguins by remote sensing. In order to gain insight into current and future populations of Antarctic penguin populations, the study has developed methods to identify the number of penguin nests to be derived as precisely as possible from the distribution of guano deposits in multispectral satellite images. In order to verify these analyzes of the satellite images, the most accurate ground truth data is necessary. Four different methods for creating such reference data were investigated and compared with each other in the project.