[1] Nicknamed the Elder (German: Ältere) to distinguish herself from her namesake great-niece, during her tenure, the Imperial court in Vienna became one of the centers of European Baroque music.

When she was ten, her education was entrusted to her paternal aunt Margherita Gonzaga, Dowager Duchess of Ferrara and Modena, who, although she never took the veil, lived in a Clarissan monastery attached to the Church of Sant'Orsola, founded by herself.

Then, negotiations began for the marriage of Eleonora with Victor Amadeus, Prince of Piedmont and heir of the Duchy of Savoy; however, the intended groom expected a large territorial gain as a dowry, which the House of Gonzaga refused.

[4] In July 1621 the widowed Holy Roman Emperor Ferdinand II sent his Privy Councillor Prince Hans Ulrich von Eggenberg to Mantua, instructing him to negotiate his marriage with Eleonora.

After obtaining the dispensation of the Holy See for the Emperor's wedding with not only a relative (ratione consanguinitatis) but also his goddaughter (ratione affinitatis spiritualis ex baptismo contractae), on 21 November the contract was signed, which repeated the provisions made by Ferdinand II's first wife, and on the same day in the Basilica palatina di Santa Barbara at the Ducal Palace, Mantua the marriage per procura was concluded.

As a wedding gift from the groom, the bride received jewelry made of diamonds and pearls worth 30,000 ducats; in addition, 18,000 florins were presented to Eleonora by the Tyrolean subjects of her husband.

They had no children, but Eleonora was able to develop a close relationship with all her stepchildren, particularly with the youngest one, the Archduke Leopold Wilhelm, in whom she formed a taste for art and literature.

[8] Like her husband, she was deeply religious and a strong supporter of the Counter-Reformation, being a benefactress of churches and monasteries and taking great interest in matters of charity.

With the blessing of Pope Urban VIII, the Empress built a monastery of Discalced Carmelites in Vienna, and in her will bequeathed 80,000 florins for prayers to be said for the salvation of her soul after her death.

She began the tradition of attending the opera and ballet performances during special celebrations in the Imperial family; the first one took place during Ferdinand II's birthday in 1625, and to this end, the Hofburg large wooden hall was built.

[14][15] A source of deep concern for Eleonora was the War of the Mantuan Succession (1628–1631), which began after the deaths of her brothers Francesco IV (1612), Ferdinando (1626) and Vincenzo II (1627) without surviving legitimate male heirs.

She was authorized by Charles II, Duke of Mantua and Montferrat to be his proxy in the negotiations of the marriage contract between his sister Eleonora and her stepson Emperor Ferdinand III,[18] which was finally signed on 8 February 1651.