The Lockport and Niagara Falls Railroad was chartered on April 24, 1834 and was completed in 1838 between the two villages. This was sometimes called the "Strap Railroad" because it consisted of straps of iron on wooden rails. For the first year the line was operated by horse power which was quickly replaced by steam engines. The original line ran through Lockport's Lowertown area, but in August 1851 the line was relocated to the upper area of the town which became the commercial center.
The Rochester and Lockport Railroad was chartered on May 15, 1837, but little was done except for some grading and bridge construction. A reorganized company, the Rochester, Lockport & Niagara Falls Railroad was granted a state charter in 1850 and the nearly 76 mile long railroad between Niagara Falls and Rochester by way of Lockport was opened to traffic in 1852. The following year the line came under the control of the New York Central railroad where is was known as the Niagara Falls Branch or the Falls Road.

The Niagara Falls Branch closely paralleled the Erie Canal for much of its length which accounts for the many stations incorporating "Basin" and "port" in their name.

The table below lists the passenger stations on the line from a 1911 timetable (above) and a January 14, 1951 timetable. Stations with no decimal point in the distance appear in the 1911 timetable but not in the 1951 version.
| Station | Distance |
| Rochester Center Park South Greece Spencerport Adams Basin Brockport Holley Fancher Albion Eagle Harbor Knollsville/Knowlesville Medina Middleport Gasport Lockport West Lockport Lockport Jct. Cambria Sanborn Walmore Suspension Bridge Niagara Falls |
0.0 0 8 10.9 14 17.9 22.5 26.1 31.4 34.3 37.1 41.2 45.7 50.7 56.6 58 61 65 67.3 70 75.7 77.5 |