Cavalcaselle was born in Legnago, Veneto. He studied at the Academy of Fine Arts in Venice. Cavalcaselle participated in the Revolution of 1848 and in the Roman Republic.
After the latter's fall, he lived in England for several years. There he published, together with Joseph A. Crowe, their first joint work, Early Flemish Painters (1856), later followed by the History of Painting in Italy (3 vol., 1864-1866). Other important works by Crowe and Cavalcaselle are The Life of Titian (London, 1876), and The Life of Raphael (London, 1883). He returned to Italy in 1857to become the chief of the art department under the Minister of Public Instruction in Rome. He was subsequently inspector of the National Museum in Florence.
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