This dissertation explores the primary sites of collecting and display commissioned by Francesco I de' Medici, Grand Duke of Tuscany (r. 1564-87). These sites ─ Francesco's studiolo in the Palazzo Vecchio and the nearby Uffizi Gallery ─ established precedents for the physical layout of newly emerging museums in early modern Europe, as scholars have suggested. But, as this dissertation asserts, Francesco's communication of authority through these sites significantly contributed to the changing expectations in the 16th century of a ruler's proper engagement with culture. Displaying objects connoting knowledge, taste, and wealth, these sites demonstrated Francesco's privileged access to such objects and his mastery over the scientific processes involved in their creation. Emphasizing the prince's knowledge, this approach contrasted with earlier rulers' reliance on images of direct military power and laid the groundwork for the merging of personal and private space that would come to characterize the full expression of absolutism across Europe. Chapter One examines the multi-faceted assertions of authority in Francesco's portraits, a strategy reflecting his embrace of images and spaces to communicate personal and political identity. Chapter Two addresses his private studiolo, which represented Francesco's participation in scientific, contemplative, and collecting activities among ruling elites. Chapter Three examines the subtle but profound shift in the meaning of the collection when, in 1583, Francesco created the Galleria degli Uffizi, a significant contribution in the history of European museums. Established independent of the prince's residence, the new museum represented Francesco's most powerful expression of cultural politics, as dignitaries visited the impressively decorated gallery and experienced first-hand its political assertions. The dissertation concludes by examining the impact of Francesco's museological precedents on other Italian rulers. Sites in Florence and Mantua demonstrate the continued attractiveness of Francesco's cultural expressions of authority to 17th-century rulers, as new expectations of a ruler's engagement with the arts emerge. Princely galleries become an increasingly common demonstration of authority, with many examples emulating the Uffizi's design. The conclusion affirms Francesco's legacy in binding the demonstration of artistic and scientific knowledge to political authority in the early Seicento.