Tuscany has some of the very best art galleries in the world. These include the Uffizi Gallery, the Bargello Museum and Santa Croce Museum.
Mediaeval era and the Renaissance is the epoch which also left the enormous quantity of art masterpieces in Tuscany. Tuscany is known for its beautiful landscapes, its rich artistic legacy and vast influence on high culture. Tuscany is widely regarded as the true birthplace of the Italian Renaissance, and has been home to some of the most influential people in the history of arts and science, such as Dante Alighieri, Francesco Petrarca, Giovanni Boccaccio, Sandro Botticelli, Raffaello Santi, Giotto, Donatello, Michelangelo, Leonardo da Vinci, Giorgio Vazari and many others and Puccini. Due to this, the region has several museums, most of which (such as the Uffizi and the Pitti Palace) are found in Florence, but others in towns and smaller villages.

In medieval period and in the Renaissance, there were four main Tuscan art schools which competed against each other: the Florentine School, the Sienese School, the Pisan School and the Lucchese School. Some of the best known artists of the Florentine School are Brunelleschi, Donatello, Michelangelo, Fra Angelico, Botticelli, Lippi, Masolino, and Masaccio.
The Sienese School of painting flourished in Siena between the 13th and 15th centuries and for a time rivaled Florence, though it was more conservative, being inclined towards the decorative beauty and elegant grace of late Gothic art. Its most important representatives include Duccio, whose work shows Byzantine influence; his pupil Simone Martini; Pietro and Ambrogio Lorenzetti; Domenico and Taddeo di Bartolo; Sassetta and Matteo di Giovanni. Siena rivalled Florence in the arts throughout the 13th and 14th centuries: the important late medieval painter Duccio di Buoninsegna (1253–1319) was a Sienese, but worked across the peninsula, and the mural of Good Government by Ambrogio Lorenzetti in the Palazzo Pubblico, or town hall, is a magnificent example of late-Medieval/early Renaissance art as well as a representation of the utopia of urban society as conceived during that period. Siena's cathedral, the Duomo, begun in the 12th century, is one of the great examples of Italian romanesque architecture. Inside is the famous Gothic octagonal pulpit by Nicola Pisano (1266–1268) supported on lions, and the labyrinth inlaid in the flooring, traversed by penitents on their knees. Within the Sacristy are some perfectly preserved renaissance frescos by Ghirlandaio, and, beneath the Duomo, in the baptistry is the baptismal font with bas-reliefs by Donatello, Ghiberti, Jacopo della Quercia and other 15th century sculptors. The Museo dell'Opera del Duomo contains Duccio's famous Maestà (1308–1311) and various other works by Sienese masters.
Unlike the naturalistic Florentine art, there is a mystical streak in Sienese art, characterized by a common focus on miraculous events, with less attention to proportions, distortions of time and place, and often dreamlike coloration. In the 16th century the Mannerists Beccafumi and Il Sodoma worked there. While Baldassare Peruzzi was born and trained in Siena, his major works and style reflect his long career in Rome. The economic and political decline of Siena by the 1500s, and its eventual subjugation by Florence, largely checked the development of Sienese painting, although it also meant that a good proportion of Sienese works in churches and public buildings were not discarded or destroyed by new paintings or rebuilding. Siena remains a remarkably well-preserved Italian late-Medieval town.

One of the distinguishing features of Renaissance art was its development of highly realistic linear perspective. Giotto di Bondone (1267–1337) is credited with first treating a painting as a window into space, but it was not until the demonstrations of architect Filippo Brunelleschi (1377–1446) and the subsequent writings of Leon Battista Alberti (1404–1472) that perspective was formalized as an artistic technique. The development of perspective was part of a wider trend towards realism in the arts. To that end, painters also developed other techniques, studying light, shadow, and, famously in the case of Leonardo da Vinci, human anatomy. Underlying these changes in artistic method, was a renewed desire to depict the beauty of nature, and to unravel the axioms of aesthetics, with the works of Leonardo, Michelangelo and Raphael representing artistic pinnacles that were to be much imitated by other artists. Other notable artists include Sandro Botticelli, working for the Medici in Florence, Donatello another Florentine and Titian in Venice, among others.