In this paper, the authors design a calculation method for landscape perception sensitivity on sight-seeing route in ecotourism destinations. The method includes 2 types of impact factors: visible perception factors and ecological perception factors. The visible perception is used to describe the perception stimulated by visual sense, which includes 3 factors, namely landscape visibility, the best viewing distance and the best viewing orientation. The ecological perception is mainly to emphasize the perception difference influenced by the ecological landscape, which also includes three factors, i. e., landscape type, resource grade and some landscapes for experiencing in them. The result shows that the calculation method for landscape perception sensitivity is a good way to quantitatively describe the eco-tourists’ perception for ecological landscapes at any locations on the sightseeing routes, and identify its spatial variation on linear units.