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The Boboli Gardens Grotta Grande, commissioned under Francesco I de’ Medici in 1583, visualizes acutely the role of materiality in the display of Third Nature philosophy. Jacopo Bonfadio described Third Nature in 1541: “For in the gardens...nature incorporated with art is made an artificer and naturally equal with art, and from them both together is made a third nature...” This analysis examines the purposeful implementation of conservative materials, including local stone, as well as water and light, as agents of Third Nature in the Grotta Grande. Light and water, as well as stone, were employed as material mediums to guide a multisensory, tactile experience that praised the extravagances of both Nature and man. The grotto’s marble sculptures, both refined and non-finito, further visualized this natural artifice. Through Francesco I’s purposeful use of materials, the Medici Grotta Grande activated a tangible Third Nature experience.