The path begins as a branch of the Main Yarra Trail, opposite St Kevin's College and Scotch College, close to Kooyong railway station (on the Glen Waverley line); between the suburbs of Toorak, Hawthorn, and Kooyong.

Near Malvern East railway station it crosses Winton Road, and shortly after this point the path diverts.

The branch to the right leads to a bridge over the Monash Freeway and railway line, giving access to Malvern East railway station or continuing on to pick up the Scotchmans Creek Trail and Malvern East Station to Centre Rd Trail.

The main Gardiners Creek Trail continues to the left, crossing the Solway Street Bridge that was rebuilt and opened in late 2011 after being washed away by flood waters in February 2011.

From Burwood Highway the path continues north-east alongside Deakin University through Gardiners Creek Reserve to Station Street, near the Box Hill Golf Club.

To remain on the Gardiners Creek Trail, riders should travel north up Old Station Street, passing immediately by the main golf club entrance, and then turn right at the quiet and dead end part of Riversdale Road.

Regardless of which route is being taken beyond Station Street, trail signage in this area is virtually non-existent, and great care needs to be taken to avoid getting lost.

Across the road, a pedestrian path, which can be used by cyclists, continues east through the Blackburn Creeklands – through Blacks Walk, Kalang Park and Furness Park to Blackburn Road; the path is unsealed through this section so slower speeds are advised.