Trams operate through the central business district on a mixed right-of-way, using dedicated traffic lanes segregated from other traffic by pavement markings. A 100 m. long segment on East Charlotte Street is the only one with trams and motor vehicles operating in mixed traffic. A 300 m. long single-track segment precedes the Fort Norfolk / Medical Center terminal.
The entire suburban segment of the line is on a dedicated right-of-way, mostly built in place of the former Norfolk Southern Railway right-of-way, parallel to Interstate I-264. All crossings are protected by gates. The segment between the Harbor Park and NSU Stations is on an extended viaduct over both the Amtrak railroad mainline and East Brambletone Avenue, State Highway 460. The longer-than-usual viaduct provides for less grades. The NSU Station is elevated.