The 501 Queen streetcar route operates between Neville Park Loop, Humber Loop and Long Branch Loop, generally in an east-west direction. It serves the Queen and Osgoode Stations on the Yonge-University-Spadina Subway. Both Queen and Osgoode Stations are accessible subway stations.

Two services are operated.  The 501 (Neville Park-Long Branch) is the main branch, and operates at all times, seven days a week. The 501 (Neville Park-Humber Loop) short-turn branch also operates at all times, seven days a week.

Westbound “501 LONG BRANCH” streetcars operate from Neville Park Loop (on the southwest corner of Queen Street East and Nursewood Road) via west on Queen Street East, west on Queen Street West, west on The Queensway, through Humber Loop, and continuing southwest and west on Lake Shore Boulevard West to Long Branch Loop (on the north side of Lake Shore Boulevard West, west of Browns Line).

Eastbound “501 NEVILLE PARK” streetcars operate from Long Branch Loop via east and northeast on Lake Shore Boulevard West, through Humber Loop, continuing east on The Queensway, east on Queen Street West, and east on Queen Street East to Neville Park Loop.

Westbound “501 HUMBER” streetcars operate from Neville Park Loop via west on Queen Street East, west on Queen Street West, and west on The Queensway to Humber Loop (on the south side of The Queensway, west of the Humber River).

Eastbound “501 NEVILLE PARK” streetcars operate from Humber Loop via east on The Queensway, east on Queen Street West, and east on Queen Street East to Neville Park Loop.

All streetcars on the route serve local streetcar stops along the route.