Home to the world's greatest collection of Italian Renaissance art, Florence's premier gallery occupies the vast U-shaped Palazzo degli Uffizi, built between 1560 and 1580 to house government offices. The collection, bequeathed to the city by the Medici family in 1743 on condition that it never leave Florence, contains some of Italy's best-known paintings including Piero della Francesco's profile portaits of the Duke and Duchess of Urbino and room full of masterpieces by Sandro Botticelli.

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The gallery is undergoing a €65 million refurbishment (the Nuovi Uffizi project) that will eventually see the doubling of exhibition space and possibly a new exit loggia designed by Japanese architect Arato Isozaki. A number of revamped rooms are open, but until the project is completed (date unknown) expect some halls to be closed and the contents of others changed.

The world-famous collection, displayed in chronological order, spans the gamut of art history from ancient Greek sculpture to 18th-century Venetian paintings. But its core is the Renaissance collection.

Visits are best kept to three or four hours maximum. When it all gets too much, head to the rooftop cafe (aka the terraced hanging garden, where the Medici clan listened to music performances on the square below) for fresh air and fabulous views.

Tuscan Masters: 13th to 14th Centuries

Arriving in the Primo Corridoio (First Corridor) on the 2nd floor, Rooms 2 to 7 – closed for renovation at the time of writing – are dedicated to pre- and early Renaissance Tuscan art. Among the 13th-century Sienese works displayed are three large altarpieces from Florentine churches by Duccio di Buoninsegna, Cimabue and Giotto. These clearly reflect the transition from the Gothic to the nascent Renaissance style. Note the overtly naturalistic realism overtones in Giotto's portrayal of the Virgin Mary and saints in the Madonna di Ognissanti.

Moving into Siena in the 14th century, the highlight is Simone Martini's shimmering Annunciazione (1333), painted with Lippo Memmi and setting the Madonna in a sea of gold. Also of note is the Madonna in trono con il Bambino in trono e otto angeli (Madonna with Child and Saints; 1340) by Pietro Lorenzetti, which demonstrates a realism similar to Giotto's; unfortunately both Pietro and his artistic brother Ambrogio died from the plague in Siena in 1348.

Masters in 14th-century Florence paid as much attention to detail as their Sienese counterparts: savour the realism and and extraordinary gold-leaf work of the Pietà di San Remigio (1360–65) by gifted Giotto pupil, Giottino.

Renaissance Pioneers

In Room 9, Piero della Francesca's famous profile portraits (1465) of the crooked-nosed, red-robed duke and duchess of Urbino are wholly humanist in spirit: the former painted from the left side as he'd lost his right eye in a jousting accident, and the latter painted a deathly stone-white, reflecting the fact the portrait was painted posthumously. Don't miss the reverse side featuring the duke and duchess etemalised with the Virtues.

Carmelite monk Fra' Filippo Lippi had an unfortunate soft spot for earthly pleasures, scandalously marrying a nun from Prato. Search out his self-portrait as a podgy friar in Incoronazione Maringhi (Coronation of the Virgin; 1439–47) and don't miss his later Madonna con Bambino e due angeli (Madonna and Child with Two Angels; 1460–65), an exquisite work that clearly influenced his pupil, Sandro Botticelli.

Another related pair, brothers Antonio and Piero del Pollaiolo, fill Room 9, where their seven cardinal and theological values of 15th-century Florence – commissioned for the merchant's tribunal in Piazza della Signoria – radiate energy. More restrained is Piero's Portrait of Galeazzo Maria Sforza (1471).

The only canvas in the theological and cardinal virtues series not to be painted by the Pollaiolos is Fortitude (1470), the first documented work by Botticelli.

Botticelli Room

The spectacular Sala del Botticelli, numbered as Rooms 10 to 14, but in fact one large hall, is one of the Uffizi's hot spots and is always packed. Of the 15 works by the Renaissance master known for his ethereal figures, La nascita di Venere (The Birth of Venus; c 1485), Primavera (Spring; c 1482), the deeply spiritual Annunciazione di Cestello (Cestello Annunciation; 1489–90), the Adorazione dei Magi (Adoration of the Magi; 1475) featuring the artist's self-portrait (look for the blond-haired guy, extreme right, dressed in yellow), and the Madonna del Magnificat (Madonna of the Magnificat; 1483) are the best known. True aficionados rate his twin set of miniatures depicting a sword-bearing Judith returning from the camp of Holofernes and the discovery of the decapitated Holofernes in his tent (1495–1500) as being among his finest works.

Leonardo Room

Room 15 displays three early Florentine works by Leonardo da Vinci: the incomplete Adorazione dei Magi (Adoration of the Magi; 1481–82), drawn in red earth pigment (removed for restoration at the time of writing); his Annunciazione (c 1475–80); and The Baptism of Christ (1470-75).

La Tribuna

The Medici clan stashed away their most precious masterpieces in this exquisite octagonal-shaped treasure trove (Room 18), created by Francesco I between 1581 and 1586. Designed to amaze and perfectly restored to its original exquisite state, a small collection of classical statues and paintings adorn its walls, upholstered in crimson silk, and 6000 mother-of-pearl shells painted with crimson varnish encrust the domed ceiling.

High Renaissance to Mannerism

Passing through the loggia or Secondo Corridoio (Second Corridor), visitors enjoy wonderful views of Florence before entering the Terzo Corridoio (Third Corridor). Rooms 25 to 32 were closed at the time of writing as part of the massive ongoing expansion and reorganisation of the Uffizi.

Michelangelo dazzles with the Doni Tondo, a depiction of the Holy Family that steals the High Renaissance show in Room 35. The composition is unusual – Joseph holding an exuberant Jesus on his muscled mother's shoulder as she twists round to gaze at him, the colours as vibrant as when they were first applied in 1506–08. It was painted for wealthy Florentine merchant Agnolo Doni (who hung it above his bed) and bought by the Medici for Palazzo Pitti in 1594.

1st-Floor Galleries

As part of the ongoing New Uffizi expansion project, the Uffizi has already added 1800 sq metres of gallery space to its vast repertoire and expansion continues into 2017. Head downstairs to the 1st-floor galleries where Rooms 46 to 55 display the Uffizi's collection of 16th- to 18th-century works by foreign artists, including Rembrandt (room 49); Rubens and Van Dyck share room 55. The next 10 rooms give to a nod to antique sculpture, before moving back into the 16th century with Andrea del Sarto (Rooms 57 and 58) and Räphael (Room 66) whose Madonna del cardellino (Madonna of the Goldfinch; 1505–06) steals the show. Räphael painted it during his four-year sojourn in Florence.

Rooms 90 to 94 feature works by Caravaggio, deemed vulgar at the time for his direct interpretation of reality. The Head of Medusa (1598–99), commissioned for a ceremonial shield, is supposedly a self-portrait of the young artist who died at the age of 39.The biblical drama of an angel steadying the hand of Abraham as he holds a knife to his son Isaac's throat in Caraveggio's Sacrifice of Isaac (1601–02) is glorious in its intensity.