The significance of the church interior– one of the few sites where all members of society (lay and religious, men and women) could interact – is only matched by our lack of precise information regarding its articulation. My book focuses on a crucial yet completely lost aspect of the Italian church interior: rood screens or tramezzi. While some previous studies have revealed evidence for screens in individual Italian churches, my book has a broader aim: to systematically and holistically investigate choir layouts to provide a comprehensive overview of sacred space in the churches of Renaissance Florence. These structures segregated social groups, sanctified hierarchical spaces, and facilitated liturgical processions. My book includes five case studies – based on the mendicant, male and female monastic, and civic contexts – which demonstrate the almost ubiquitous presence of screens, each case presenting unpublished archival documentation and new architectural reconstructions whilst elucidating wider art-historical issues such as gender, patronage, and social class. Despite their prevalence and diverse functionality, tramezzi were widely destroyed in the later sixteenth century and their accompanying choir stalls were relocated to areas behind the high altar. My book situates Florence, and Italy more generally, within a broader context of pan-European societal and religious change, arguing that both the Protestant and Catholic traditions experienced a reorientation of religious belief, shift in ideological focus, and change in liturgical practice which found variable expression in architectural form.