Mikveh Israel Cemetery serves as the oldest record of a permanent Jewish community in Philadelphia. It was established by merchant Nathan Levy following the death of his son in 1740.

An observant Jew, Levy sought a parcel of land on which to bury his son in accordance with Jewish law. Thomas Penn, son of William Penn and royal proprietor of Pennsylvania, allowed Levy to purchase a small private space surrounded by a "fence of boards . . . [in] an area characterized by woortleberry bushes and cows grazing in open fields." The cemetery, on the north side of Spruce between Eighth and Ninth Streets, evolved into a communal burial ground for colonial Philadelphia Jews.

Jewish traders periodically visited the Delaware Valley before William Penn's landing in 1682. Originally, some people from Dutch-held New Amsterdam colony (now New York) were drawn to the lucrative fur trade, along what was then known as the South River (now the Delaware River). Migration between that area and Philadelphia swelled after the establishment of Penn's colony and its promise of religious tolerance. Nathan Levy was a member of this group, eventually forming the first important Jewish company in Philadelphia: the Levy & Franks import/export merchants.

Ownership of the cemetery was transferred to the Mikveh Israel Congregation in 1828. Known as the "Synagogue of the American Revolution," the congregation is the oldest continuous synagogue in the United States

Eminent citizens interred at the cemetery include the great American patriot Haym Salomon, a member of the Sons of Liberty and a spy during the Revolution; Aaron Levy, influential pioneer and founder of Aaronsburg; and members of the Gratz and Phillips families. The cemetery also serves as the resting place of Jewish veterans of the American Revolution and the War of 1812.

Designated a National Historic Shrine in 1956, the cemetery is accessible from the intersection of Ninth and Locust Streets.