The southern half of NY 65 passes through mostly rural areas, while its northern section traverses densely populated portions of Monroe County.

NY 65 originally began at what is now US 20A in Honeoye when it was assigned as part of the 1930 renumbering of state highways in New York; however, it was truncated to its current southern terminus in the late 1930s.

[6] The route continues onward, passing alongside the western edge of Mendon Ponds Park as it enters the town of Pittsford.

At the northern edge of the park, NY 65 curves to the northeast and passes over the New York State Thruway (Interstate 90).

From this point northward, the route heads through much more developed areas, beginning with a series of housing tracts situated in the 1.5 miles (2.4 km) between Calkins and Jefferson Roads, the latter carrying NY 252.

It continues on past NY 252, taking on a more northerly routing as it crosses over the CSX Transportation-owned West Shore Subdivision railroad line and the Erie Canal and passes through densely populated portions of Pittsford and Brighton.

[8][9] The route heads due northward for just under 1 mile (1.6 km), paralleling I-590 and passing The Harley School before veering to the northeast ahead of an intersection with Elmwood Avenue.