The first U.S. Consulate in this consular district was established in the port city of Leghorn (Livorno). U.S. Consular Agent Philip Felicchi was appointed on May 29, 1794. The Government of the Grand Duchy of Tuscany did not recognize U.S. Consular Agents posted in Florence; thus the first U.S. Consular Agent to serve Florence was Vice Consular Agent James Ombrosi, who was under mandate from the U.S. Consulate at Leghorn (Livorno). Ombrosi was accredited on May 15, 1819.

• James Ombrosi 1819-1849 Consular Agent (Consul after 1825)
• Edward Gamage 1849-1852 Consul of Florence & Livorno
• Francis Lance 1852-1854 Commercial Agent
• Amasa Hewins 1854-1855 Commercial Agent
• 1856-1857 Post Vacant
• Edward J. Mallett 1858-1861 Commercial Agent (Consul General after 1860)
• T. B. Lawrence 1861-1869 Consul General (Florence: Capital of Italy 1865-1870)
• J. Lorimer Graham, Jr. 1869-1876 Consul General (Consul after 1871)
• J. Schuyler Crosby 1876-1882 Consul
• William L. Welsh 1882-1887 Consul
• Isaac R. Diller 1887-1891 Consul
• James V. Long 1891-1893 Consul
• Charles B. Davis 1893-1897 Consul
• Edward C. Cramer 1897-1903 Consul
• Francis B. Keene 1903-1905 Consul
• Jerome A. Quay 1905-1910 Consul
• Leo J. Keena 1910-1914 Consul
• Frederick T. F. Dumont 1914-1919 Consul
• W. Roderick Dorsey 1920-1923 Consul
• Joseph E. Haven 1923-1937 Consul
• John R. Putnam 1937-1941 Consul General
• 1941-1944 Consulate closed during World War II
• Walter W. Orebaugh 1944-1948 Consul
• George P. Waller 1948-1950 Consul
• Charles S. Reed II 1950-1954 Consul General
• Richard M. Service 1954-1956 Consul
• William D. Fisher 1956-1958 Consul
• Merritt N. Cootes 1958-1966 Consul General
• Joseph C. Wheeler 1966-1970 Consul General
• William J. Barnsdale 1970-1972 Consul General
• Robert C. F. Gordon 1972-1978 Consul General
• Donald A. Johnston 1978-1982 Consul General
• Frederick H. Hassett 1982-1985 Consul General
• Diane Dillard 1985-1989 Consul General
• Virginia Morris 1989-1990 Consul
• Marisa R. Lino 1990-1993 Consul General
• Sue H. Patterson 1993-1996 Consul General
• Louis A. McCall 1996-1998 Consul General
• Hilarion Martinez 1998-2001 Consul General
• Daria M. dePierre-Hollowell 2001-2002 Consul General
• William W. McIlhenny 2002-2005 Consul General
• Nora B. Dempsey 2005-2008 Consul General
• Mary Ellen Countryman 2008-2011 Consul General
• Sarah C. Morrison 2011-2014 Consul General
• Abigail M. Rupp 2014-2017 Consul General
• Benjamin V. Wohlauer 2017-2020 Consul General