Early sixteenth century saw the emergence of printed sources of polyphony alongside music manuscripts, and the new technologies applied to the creation of music books inevitably brought change to their material properties and dissemination. This thesis examines the effects of printing technology to the layout of music, text, and images in books of polyphony from the first half of the sixteenth century. The research is a part of an AHRC-funded project 'Production and Reading of Music Sources, 1480-1530' on systematic analysis on the production and use of polyphonic music manuscripts and prints from the late fifteenth century to the early sixteenth century. First part of the thesis (Chapter 2) re-evaluates the music printing techniques used during the early sixteenth century, followed by a bibliographical analysis of the most common formats and layouts of the surviving sources: large choirbooks, small choirbooks, and partbooks (Chapters 3, 4, 5). The analysis on these chapters focuses on the structure of the books and the printing materials used in their making, traces printing house procedures evident from the layout in order to determine visual hierarchies on the presentation of music, text, and decorative devices, and how these hierarchies would change depending on the geographic location and the technological requirements. Chapter 6 analyses later changes made to printed books, both in the printing house and by the later readers of the books, with a case study based on Andrea Antico's Liber quindecim missarum printed in 1516.A closer observation of the printing techniques suggests further sub-categories on the main division between sources printed from woodcuts or from movable type. In particular, the methods used for printing from woodcut can be divided into three categories: block-book printing, the use of movable type along the woodcuts, and separating the notation and staves to two separate blocks for multiple impressions. These subdivisions allow further understanding of the printing house processes, and the individual approaches taken by the makers of these books. The relatively small readership of polyphonic sources was receptive for variety in terms of format and layout. While a portion of printed books share same repertory, they are rarely identical: the differences in choice of technique and its implementation on the layout and format bring an individual touch to the publications, reflecting the diversity present in the surviving sources of manuscripts and prints from the first decades of the sixteenth century.