The Biblioteca Ambrosiana is a historic library in Milan, Italy, also housing the Pinacoteca Ambrosiana, the Ambrosian art gallery. Named after Ambrose, the patron saint of Milan, it was founded in 1609 by Cardinal Federico Borromeo, whose agents scoured Western Europe and even Greece and Syria for books and manuscripts.

During Cardinal Borromeo's sojourns in Rome, 1585–95 and 1597–1601, he envisioned developing this library in Milan as one open to scholars and that would serve as a bulwark of Catholic scholarship in the service of the Counter-Reformation against the treatises issuing from Protestant presses. To house the cardinal's 15,000 manuscripts and twice that many printed books, construction began in 1603 under designs and direction of Lelio Buzzi and Francesco Maria Richini. When its first reading room, the Sala Fredericiana, opened to the public on 8 December 1609 it was, after the Bodleian Library in Oxford, the second public library in Europe.

One innovation was that its books were housed in cases ranged along the walls, rather than chained to reading tables, the latter a medieval practice seen still today in the Laurentian Library of Florence. A printing press was attached to the library, and a school for instruction in the classical languages.

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