In this article, we use eye-tracking methods to analyze the differences in spatial ability between geographers and non-geographers regarding topographic maps, as reflected in the following three aspects: map-based spatial localization, map-based spatial orientation, and map-based spatial visualization. We recruited 32 students from Beijing Normal University (BNU) and divided them into groups of geographers and non-geographers based on their major. In terms of their spatial localization ability, geographers had shorter response times, higher fixation frequencies, and fewer saccades than non-geographers, and the differences were significant. For their spatial orientation ability, compared to non-geographers, geographers had significantly lower response times, lower fixation counts and fewer saccades as well as significantly higher fixation frequencies. In terms of their spatial visualization ability, geographers’ response times were significantly shorter than those of non-geographers, but there was no significant difference between the two groups in terms of fixation count, fixation frequency or saccade count. We also found that compared to geographers, non-geographers usually spent more time completing these tasks. The results of this study are helpful in improving the map-based spatial ability of users of topographic maps.