The Louisville & Nashville Railroad Company was chartered in Kentucky on March 5, 1850 for the purpose of building a railroad line between Louisville, Kentucky, and Nashville, Tennessee. The first train to travel the railroad from Louisville to Nashville ran on October 27, 1859, and regular trains began running a few days later. During the Civil War, the L&N lay between Union and Confederate Lines and served both sides at various times. The railroad began expanding after the war ended, and eventually linked such cities as Cincinnati, Ohio, Pensacola, Florida, New Orleans, Louisiana, St. Louis, Missouri, Memphis, Tennessee, Birmingham, Alabama, and Mobile, Alabama. After transporting both freight and soldiers during World War II, the L&N continued to expand, purchasing new track and offering streamlined passenger service with such trains as The Dixie Flyer, The Georgian, and The Hummingbird. In 1969, the L&N acquired part of the Chicago & Eastern Illinois Railroad, which gave it access to important midwestern rail centers. In 1971, however, Seaboard Coast Line Industries bought the L&N, and it was officially merged into the Seaboard System Railroad on December 31, 1982. In 1986, the Seaboard System and the Chessie System merged into CSX Transportation, which continues to operate trains on what was once L&N track today.

The L&N Station in Knoxville, Tennessee was constructed in 1905 after the purchase of the the Atlanta, Knoxville and Northern Railroad, which connected Knoxville to the Western and Atlantic at Marietta, Georgia. The station was built to rival the Southern Terminal building on Depot Street. The L&N Station was designed by Richard Montfort. Author James Agee described the station in his book A Death in the Family, which takes place in Knoxville in 1915. The L&N Station was rennovated for the 1982 World's Fair in Knoxville and was then turned into a magnet school, L&N STEM Academy.