The Virginia Museum of Transportation, located in downtown Roanoke, is housed in the former Norfolk & Western Railway Freight Station. Built in 1918 in the Colonial Revival style, it is listed on the National Register of Historic Places and Virginia Landmarks Register. The museum contains around 2,500 transportation-related objects, over 50 locomotives and trains cars, and the largest collection of diesel locomotives in the American south. The museum also has planes, vintage automobiles and other transportation artifacts. It is part of with the Virginia Rail Heritage Region, an organization formed in 2010 dedicated towards celebration the region's rail history.

Located in Roanoke, Virginia, The O. Winston Link Museum is housed in the former Norfolk & Western Railway passenger train station, which was built in 1905 and redesigned in 1949 by architect Raymond Loewy. The building is part of the Norfolk and Western Railway Company Historic District. The museum is named after photographer O. Winston who, in the 1950s, documented the end of the Norfolk & Western steam train era. He recorded the trains and the communities they served in photographs and films, all of which are located in the museum. Link's most well-known photograph is "Hotshot Eastbound" (see below). The museum opened in 2004.

The History Museum of Western Virginia is located in downtown Roanoke and is dedicated towards promoting the history of southwestern Virginia. It is housed on the third floor of the Center in the Square building. The Watts Library and the administration offices of the Historical Society of Western Virginia and are located inside the museum. The museum receives over 25 thousand visitors per year. It contains thousands of artifacts and other historical materials from western Virginia. These include photographs, documents, books, and maps. A notable item is a letter written by Thomas Jefferson describing his scientific observations of the the three mountains, called the Peaks of Otter, that overlook the town of Bedford. The museum is currently undergoing a $3 million dollar restoration.