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The Uffizi Gallery has been altered several times in its four centuries of life. The Tribune by Buontalenti (1584), today Room 18, formed the first ordered nucleus of the museum, together with other adjacent rooms: the Room of Scientific Instruments (today Room 17), the Terrace of Geographical Maps (today Room 16) and the five smaller rooms created by Francesco I's brother and successor, Ferdinando I, to display an arms collection (today Rooms 19-23). The Room of Madame (today Room 24) the Room of Miniatures), where the Grand Duchess kept her jewellery and where "idols" (small classical bronzes) and miniatures were displayed, stood on the Eastern side of the end of this first corridor. Although they now mostly contain paintings, these rooms still maintain their suggestive sixteenth century atmosphere.

Ferdinando II had the ceilings of the second and third corridors completed with frescoes dedicated to the glories of Florence in the 17th century while Cosimo III used these rooms to exhibit recently added jewels to the grand-ducal collections. The Vestibule (now the exit on the staircase by Buontalenti) was therefore created along the long West Corridor, to contain archeological discoveries and ancient inscriptions, the Porcelain Room (today Room 34, Veronese), the Room of the Medals (today Room 41, Rubens), with over 30.000 exhibits, and the Room of Selfportraits (today Room 35, Tintoretto): this last collection was later moved into the Vasari Corridor.

The drawings collected by Cardinal Leopoldo, the main nucleus of the Gallery of Prints and Drawings, the miniatures and objects in pietra dura were also stored in the Gallery while awaiting definite arrangement. The collection of classical sculpture was also growing, thanks to the acquisition of some really exceptional examples like the Hermaphrodite, Cupid and Psyche, the Wrestlers and the Medici Venus. By 1689 about 102 busts and 72 statues were on display along the corridors. Their first official restorer, the sculptor Ferrata, was lodged in Palazzo Vecchio in order to be as close as possible to the works he had to restore.

The Gallery of the Uffizi certainly did not lose any of its importance during the reign of the Lorraine family (1737-1860) for acquisitions of works of art still continued even though some unfortunate errors were made, like the sale of the Medici Armoury (1775) and the reduction of the majolica collection.

Some of the most important innovations were however of administrative type. Giovanni Bianchi, the official "Custodian", printed the first guide to the museum (1759), the Room of Niobe (West Corridor, architect Gaspare Maria Paoletti) and the Room of Jewels, today Room 24, Miniatures (1782, architect Zanobi del Rosso), were arranged, a director was nominated (1765, Giuseppe Bencivenni Pelli, with Luigi Lanzi as assistant), the Royal Guards were put in charge of the security of the paintings (1793) and all the works were labelled to make it easier for visitors to identify them (1795).

Stimulated by the new principles of Illuminism, Grand Duke Pietro Leopoldo of Lorraine started (1780-82) to arrange the collections in a scientific-didactic order and transferred some of the sections in the Uffizi Gallery to other galleries like the Academy and the Opificio delle Pietre Dure.

Between the 18th and 19th centuries, entire collections were transferred elsewhere to create new museums: La Specola (scientific instruments), the Egyptian Museum (1852), the Etruscan Museum (1870) and the Bargello (1865), dedicated to 15th-16th century sculpture. A section of paintings was detached to create the Gallery of Modern Art at the Pitti Palace, the paintings of Fra Angelico were sent to St. Mark's, a whole set of paintings was sent to Lucca.

Alterations carried out in this century have led to the creation of the Room of the Primitives by the architects Scarpa, Michelucci and Gardella (1953-56), the restoration of the church of San Pier Scheraggio which Vasari had incorporated in the Palace (1971, architect Bemporad), a new arrangement of the Lippi (1973-74, Room 8), Botticelli (1978, Room 10-14) and Leonardo Rooms (1979, Room 15). About 1.700 paintings are exhibited at present, while another 1.000 are displayed (and can be viewed on request) in the new Storerooms created in 1978. The project for the "Great Uffizi" is still in the throes of being carried out; the plan dates from 1964 and will also include a new entrance from Piazza Castellani and a substantial increase in space for exhibitions as well as for equipment and other facilities. Thus the rooms on the first floor, which were occupied by the State Archives until only a few years ago, will be added to the historic 45 rooms on the second floor.