Understanding human behavior in emergency evacuation is a significant issue for layout optimization, crowd management and rescue. In this study, we conducted a series of controlled experiments to study choice behavior of pedestrians considering environment factors (e.g. occupant initial distribution, auditory information, and building layout). We found it was non-symmetrical for pedestrians’ exit selection and aisle selection in the room. And there was a strong positive relation between intermediate exit choice and destination choice. Pedestrians’ final destinations had significantly effects on evacuation route and intermediate facilities usage. When the final destination was uncertain, the factor of building layout performed more effect than occupant initial distribution. Pedestrian psychology, following the crowd, had a major influence on pedestrians’ exit stairs selection, especially when pedestrians were in non-limited visual field environment. Bifurcation point, where a row of pedestrians split into two streams with diametrically opposite movement directions, was a quite biased away from the side of exit. The conclusions are expected to provide valuable advice for crowd management and optimization design such as aisle-seating layout.