Wayfinding is the spatial knowledge about one’s current location, destination, and the spatial relation between them. Wayfinding problems threaten people’s sense of well-being, and cause loss of time and money. Designers and planners can improve wayfinding when they understand how physical environmental factors affect people’s wayfinding performance. This study explores the effect of personal and physical environmental characteristics on wayfinding performance. The personal characteristics include gender, age, and familiarity. The physical environmental characteristics include plan layout complexity, physical differentiation and its components vertical and horizontal differentiation. 166 volunteers explored 18 virtual environments and completed wayfinding tasks, such as sketching, and direction estimation. The results showed that the Simple layouts, Higher Physical Differentiation, Vertical or Horizontal differentiation yielded better wayfinding performance than Complex layouts, Lower Physical differentiation, and No Vertical or Horizontal differentiation. Males performed better than Females, and performance improved with Familiarity.